Laboratory Manual Fall 2014

Rhode Island College Biology 335
Contents
Revised Fall 2014
1. Safety in the Laboratory
2. Measurements and Computations
3. The Cell: Transport Mechanisms and Cell Permeability
13
4. Introduction to iWorx
15
5. Physiology of Skeletal Muscle
27
6. Skeletal Muscle Physiology: Computer Simulation
39
7. Reflexes
41
8. Cardiology with a Vertebrate Heart
51
9. Electrical Properties of the Heart
65
10. Circulatory Physiology
79
11. Mechanisms of Breathing
89
12. Restrictive and Obstructive Lung Disease
103
13. Basal Metabolic Rate
111
14. Renal Physiology The Function of the Nephron
123
Page 1
Revised Fall 2014
Page 2
Biology 335 Human Physiology: Safety
Laboratory #1
Safety in the Laboratory
discuss your ideas with the instructor at the time they
The laboratory is a safe place to work and study. It remains
occur to you or before leaving the laboratory.
safe when the individuals working there practice the conventional rules of laboratory safety. Laboratory work frequently
The laboratory bench must be clean and organized, and
requires the use of reagents, equipment, and organisms that
other extraneous items are a safety hazard when stacked
are potentially dangerous for all personnel in the laboratory.
on the surface of the bench. Remove unnecessary items
On entering the laboratory, you assume
from the bench top and place them in
responsibility for your own safety and
The keys to working safely in
a secure storage area. The cabinet in
the safety of your neighbors. The adany laboratory are organization, the bench pedestal is a good place to
vice that follows represents accepted
store calculators, purses and other
procedure and describes the behaviors
neatness and being prepared!
valuables.
assumed to be characteristic of anyone
working in a laboratory.
The laboratory in which you
will be working is a general purpose teaching laborato Good laboratory work requires advanced preparation on
ry. Instructors and students from different courses
the part of all that are present. The instructions and
make use of the room. As a result, a wide variety of
directions for a specific activity require careful study
substances and equipment is constantly being moved
beforehand. You need to know: 1) What to do, 2) How
into and out of the laboratory. Materials remain and
to do it, and 3) Why it is to be done in the prescribed
form residue that is potentially pathogenic, capable of
manner. Thorough preparation improves the quality,
causing personal injury, and likely to soil or damage
efficiency, and safety of your work. Prepare a set of
your belongings. Check the bench top as a precautionnotes for the materials and procedures organized in the
ary measure before assembling the materials needed for
form of a flow chart. Highlight notes that describe safethe laboratory activity. Keep your hands away from
ty procedures.
your mouth. Do not eat or drink in the laboratory,
and never pipette by mouth suction.
Instructors usually provide a briefing before the start of
a laboratory exercise. Unless the instructor tells you
When gathering materials from central supply areas,
otherwise, it is advisable to wait for that commentary
label the containers for their transport beforehand.
before beginning your work. The briefing provides the
Read the labels on the stock containers TWICE before
opportunity for: describing changes in instructions,
taking what you need. Immediately replace caps, stopdemonstrating techniques, explaining the procedures for
pers, and covers for all containers. The stock is to rethe proper use of instruments, and highlighting safety
main in the supply area. Excess amounts of media and
precautions. Be sure that you understand, and follow
reagents are not to be returned to stock supplies because
exactly, the special
contamination of the stock supply may result from
procedures for the
Please note that the material in
this process. Materials and equipment are to remain
correct disposal of
this manual is a compendium of
in the laboratory at all times.
hazardous materiexercises originating from multiple
als, biological
A general knowledge of safety procedures is
sources. The computer simulawastes, and body
tion laboratories are modifications inadequate. You must know what to do in a particufluids. The briefing of exercises contained within the
lar set of circumstances. Quick action is vital when
is a chance for you PhysioEx Laboratory Manual.
accidents occur that result in materials entering the
to ask questions
eyes, fire, broken glass, chemical spills, or injuries
Contributors to this manual inabout equipment
clude Frank Dolyak, Kenneth Kin- to the skin. Therefore, thoroughly study the proceand procedures that sey, Edythe Anthony, Jerome
dures that are to be followed for each of the condiyou dont undertions listed below.
Montvilo, Eric Hall and others.
stand. Making
Eye Injury. Go immediately to the eye wash station
changes in the procedures and using materials other
and flush the material from the eyes. When
than those described in the instructions can be disasone eye is affected, tilt the head to prevent
trous. On the other hand, changes based on new inthe stream of water from introducing the
sights can be very valuable because they may increase
material into the other eye. Continue the
the effectiveness of an exercise. Therefore, be sure to
Revised Fall 2014
Page 3
Biology 335 Human Physiology: Safety

procedure until directed to stop. Individuals in
the immediate area are to clear a path to the
eye wash station and take the initiative to lead
the injured person to the wash station. NOTE:
If you wear contact lenses and also have a
framed set of lenses, it is advisable to wear the
framed lenses in the laboratory.
as you would like to find it. Your colleagues and

instructors appreciate your cooperation. Thank
you!
Fire. When entering the laboratory for the

first time, determine the shortest exit routes
from your laboratory station to the adjoining
hallways. Locate the exits leading from the hallways to the
outside of the building. If the laboratory has fire extinguishers and fire blankets, the instructor will explain their proper
use and show you their locations.
Broken glass. To pick up broken glass use a dust pan and
brush. Gather together small slivers and chips by using a
crumbled wad of wet paper toweling or wet cotton and laboratory tongs. Discard the glass into the special container designated by the instructor.
Chemical spills. Clear the

area immediately and tell
the instructor. Promptly
flush the area with water
when spilled material contacts your personal belongings, clothing or skin.
CAUTION! Sometimes alternative procedures are required,
and the instructor will describe them during the laboratory
briefing.
Eye Wash Station
Skin injuries. Immediately report accidents that puncture,

cut, abrade or burn the skin. In each laboratory, there is a
first aid kit for the immediate treatment of minor injuries.
Any occurrence is significant, and the injured person should
consult the college Health Service.
Long hair, long or baggy sleeves, large bracelets, and

long dangling necklaces are safety hazards. When necessary, tie back long hair and roll up long baggy sleeves.
Store jewelry and other personal items in a secure area.
Sandals should not be worn in the laboratory. They do
not provide adequate protection against sharp objects or
chemical spills.
Before leaving the laboratory, be sure to clean and restock the individual bench area. Arrange the materials
and equipment as they were initially, and discard wastes
into the proper containers for disposal. CAUTION!
Disposal of reagents, biological wastes and body fluids
in a casual manner endangers yourself and those around
you. Unless told to do otherwise, solid waste is not
discarded into the sink or solution poured down the
drain. You should strive to leave the laboratory station
Revised Fall 2014
First Aid Kit and Fire Blanket
Page 4
Biology 335 Human Physiology: Measurements and Computations
Laboratory #2
Measurements and Computations
Objectives
1.
2.
3.
4.
HowmanyEnglishunitsinam,lorg?
Review the metric system.

Practice converting between different metric units
Review simple computations such as using
ratios and proportions.
Review the use of significant figures.
1.093yd
3.279
39.37in
0.264gl
1.057qt
2.114pt
0.0022lbs
0.035oz
Table 1. Metric and English Equivalents
I. The Metric System

units and their interchangeability.
At this point in your science education you should be
intimately familiar with the standard units of measurement used throughout the world. You should also possess a concept of the relationship between these international standards and the measures commonly used in the
USA. This latter is especially important for health care
providers who must translate a patients ideas of measure
into these standard measurements for recording in the
patients records. In fact, the measurement and administration of medications is always done in metric units.
When you pick up a prescription at the drug store it will
be measured in international units and if you are administering medications in a clinical setting you will also be
using international units. Of course the international
standard we are discussing is the metric system which is
based on the decimal system where the units are related
to each other by powers of 10. The following prefixes
have a constant meaning and are in general use throughout the metric system:
Kilo (k) =
Centi (c) =
Milli (m)=
Micro () =
Nano (n)=
thousand (times) [103]

one-hundredth [10-2]
one-thousandth [10-3]
one-millionth [10-6]
one-billionth [10-9]
B. Mass Units
Instead of weight (pounds and ounces), the metric
system expresses mass in grams and multiples thereof.
See the conversion table on the next page. It might be
more flattering to think of your weight in kg (a 200 lb
man or woman has a mass of about 91 kg), but your
actual mass is the same regardless of the units you use
to express it.
C. Volume Units
The metric unit of volume is the liter (l; slightly more
than a quart) and its multiples. See the table on the
next page.
D. Time
Seconds are divisible into milliseconds (ms) and microseconds (s). There are 1000 ms in 1 s and
1,000,000 s in 1 s.
E. Electrical potential difference
Volts (V) are divisible into millivolts (mV) and microvolts (V).
A. Length Units
Instead of the yards, feet and inches of the English system, the metric system expresses the length of an object
in meters, centimeters and millimeters. Still smaller
units are micrometers and nanometers. For comparison,
a human erythrocyte is about 7 micrometers in diameter.
Table 1 presents some commonly used metric length
F. Temperature Scales
In the Fahrenheit scale (still used in the USA), water
boils at 212 F, and freezes at 32 F, and the range
comprises 180 units. In the Celsius (centigrade) scale
the boiling point of water is 100 C and the freezing
Continued on page 7.
Revised Fall 2014
Page 5
ToConvert:
To:
Mul plyby:
ngornlornm
gorlorm
ngornlornm
mgormlormm
ngornlornm
gorlorm
ngornlornm
kgorklorkm
gorlorm
ngornlornm
1000 (1X103)
gorlorm
mgormlormm
0.001 (1X103)
gorlorm
gorlorm
gorlorm
kgorklorkm
0.000000001 (1X109)
mgormlormm
ngornlornm
1000000 (1X106)
mgormlormm
gorlorm
1000 (1X103)
mgormlormm
gorlorm
0.001 (1X103)
mgormlormm
kgorklorkm
0.000001 (1X106)
gorlorm
ngornlornm
1000000000 (1X109)
gorlorm
gorlorm
1000000 (1X106)
gorlorm
mgormlormm
gorlorm
cgorclorcm
100 (1X102)
gorlorm
dgordlordm
10 (1X10)
gorlorm
kgorkgorkm
0.001 (1X103)
kgorklorkm
ngornlornm
1000000000000 (1X1012)
kgorklorkm
gorlorm
kgorklorkm
mgormlormm
kgorklorkm
gorlorm
Revised Fall 2014
0.001 (1X103)
0.000001 (1X106)
0.000000001 (1X109)
0.000000000001 (1X1012)
0.000001 (1X106)
1000 (1X103)
1000000000 (1X109)
1000000 (1X106)
1000 (1X103)
Page 6
point is 0 C, with a range of 100 units. Thus one Celsius unit of temperature is larger than one Fahrenheit
unit, specifically 180/100 or 9/5 greater.
B. Proportion
To convert a Fahrenheit temperature to Celsius, use the A proportion is a statement of the equality of two ratifollowing formula:
os and can be expressed in this way:
To convert a Celsius temperature to Fahrenheit, use the

following formula:
We state that a is to b as c is to d. If the numerical
value of three of the four terms is known, the fourth
can be determined by the following formula (this is
called cross multiplying):
H. Problems
When you convert from a larger unit to a smaller unit
move the decimal to the right. When you convert
from a smaller unit to a larger unit move the decimal
to the left:
arge
mall
For instance, assume that for every liter of blood

pumped by the heart, 300 mL of this blood goes to the
kidneys. When the heart pumps harder and puts out 3
liters of blood in the same amount of time, you wish to
know how much blood goes to the kidneys (assuming
no independent change in kidney flow). You would
solve the problem by using the following proportion:
Solve the problems found on the accompanying worksheet.
II. Computations and Presentation of Data

A. Ratios
A comparison of two numbers is called a ratio. For
instance, if a tank of frogs contains 150 green frogs
and 75 brown frogs, the ratio of green to brown frogs
is 150 to 75, which may be written 150/75 or 150:75.
This ratio is a fraction which should be reduced to
the lowest common values (2/1 or 2:1). [It may be
expressed as a single number by dividing it out (in this
case 2). A denominator of 1 is then assumed.]
When two numbers are expressed as a ratio their units
must be the same. For instance, if a rabbit has a mass
of 1.5 kg and a frog has a mass of 100 g, one could
express the ratio of their masses, but only after converting one of the measurements to the units of the
other. Since 1.5 kg = 1500 g, we can express the ratio
in grams as 1500: 100 or 15:1. Conversely, we could
convert g to kg, getting 0.1 kg for the weight of the
frog. The ratio would be the same in kg, i.e. 1.5:0.1 =
15:1.
Revised Fall 2014
C. Calculation of an Arithmetic Mean

Performing a physiological or any scientific experiment involves the collection of a great deal of numerical data. After collecting the data, it is usually necessary to determine the significance of the data. Often
we want to know if the experimental procedure produced an effect which was different from a control
procedure. In order to compare the experimental
group of data with the control group it is first necessary to condense the data, that is, calculate the arithmetic mean. The arithmetic mean is simply the average value of the group data and is calculated as:
Page 7
Biology 335 Human Physiology: Measurements
X= Nx
Where:
X

x
= sum
= each individual datum
= number of data values in the group
Experimental thyroid mass (mg):

157
174
205
180
181
168
x = 1065
= 1065 6 = 177.5 mg
177.5 mg is the experimental mean.

The next step would be to compute the mean of the
control group, in the same way:
2. Control thyroid mass (mg):
160
154
190
142
201
179
Revised Fall 2014
= 1026 6 = 171 mg
171 mg is the control mean.
For instance, a group of six rats was given thiouracil

for two weeks in order to repress the functioning of the
thyroid gland. A valid question could be raised, Does
administration of thiouracil cause atrophy of the thyroid gland? In order to answer this, you could determine the mass of the thyroid gland from each of the six
experimental rats. These data would have to be compared with similar data from a control group of rats.
The first step would be to compute the arithmetic mean
of the experimental data and compare it with the arithmetic mean of the control data, as follows:
1.
= mean
x = sum of all individual data of one group

N
x =1026
By comparing the two arithmetic means, we would

conclude that there was no atrophy of the thyroid
gland caused by the procedure.
Suppose we had found a larger difference between the
experimental and control means. Let us assume a different set of control values, such that the control mean
is 300 mg. In this case, the experimental mean of
177.5 mg is so much smaller than the control mean
that we might be justified in concluding that atrophy
of the thyroid had been produced by the thiouracil.
In many biological experiments the results are not so
clear cut. We cannot tell by simply inspecting the
means, whether there is a significant difference between the experimental mean and the control mean. In
those cases it is necessary to use statistical procedures
which will tell us the probability that there is, or is not,
a significant difference between the two means.
D. Problems
Complete the problems included on the following
worksheet.
III. Graphing
The data gathered in the Human Physiology laboratory
is fairly simple and does not require complex graphing
techniques or expertise. At the same time there are
some things that a student should be aware of when
graphing data. The following are general rules that
should be used whenever you create a graph (and must
be followed when graphing data in this class).
1. You must have a title at the top of the page
2. You must properly label the X and Y axes
3. The X axis (horizontal) is the independent variable (Time or temperature in the Physiology laboratories)
4. The Y axis (vertical) is the dependent variable
(what you measured and recorded as your data)
5. Labels MUST always include units. Examples:
Time (min), Heart rate (Beats/min)
6. Please make every effort to fit the data optimally
to the range of each axis.
7. Use the space provided on the graph paper to optimally display your results
Page 8

8.
9.
Use the appropriate graph type

For continuous independent variables (time) use
a line graph
10. For discrete independent variables (temperature)
use a bar graph
11. When a graph contains two discrete data sets on
one graph make sure there is an appropriate label
or legend to make it clear.
12. Finally, follow the guidelines provided in the
manual.
See the provided examples.
IV. Significant Figures

All measurements are approximationsno measuring device can give perfect measurements without
experimental uncertainty. By convention, a mass
measured to 13.2 g is said to have an absolute uncertainty of 0.1 g and is said to have been measured to
the nearest 0.1 g. In other words, we are somewhat
uncertain about that last digitit could be a 2; then
again, it could be a 1 or a 3. A mass of 13.20 g
indicates an absolute uncertainty of 0.01 g.
The number of significant figures in a result is simply
the number of figures that are known with some degree of reliability. The number 13.2 is said to have 3
significant figures. The number 13.20 is said to have
4 significant figures.
A. Rules for deciding the number of significant
figures in a measured quantity:
(1) All nonzero digits are significant:
1.234 has 4 significant figures,
1.2 has 2 significant figures.
(2) Zeroes between nonzero digits are significant:

1002 has 4 significant figures,
(4) Zeroes to the right of a decimal point in a number

are significant:
0.023 has 2 significant figures,
(5) When a number ends in zeroes that are not to the

(3) Zeroes to the left of the first nonzero digits are not
right of a decimal point, the zeroes are not necessarily
significant; such zeroes merely indicate the position of
significant:
the decimal point:
0.001 has only 1 significant figure,
Revised Fall 2014
190 may be 2 or 3 significant figures,

50,600 may be 3, 4, or 5 significant
Page 9

figures.
last remaining digit is left as it is. For example,
12.4 is rounded to 12.

The potential ambiguity in the last rule can be avoided
by the use of standard exponential, or scientific, notation. For example, depending on whether 3, 4, or 5
significant figures is correct, we could write 50,600
(3) If the digit to be dropped is 5, and if any digit folcalories as:
lowing it is not zero, the last remaining digit is increased by one. For example,
5.06 104 calories (3 significant figures)
5.060 104 calories (4 significant figures), or
5.0600 104 calories (5 significant figures).
B. Rules for mathematical operations
(4) If the digit to be dropped is 5 and is followed only

by zeroes, the last remaining digit is increased by one
In carrying out calculations, the general rule is that the
if it is odd, but left as it is if even. For example,
accuracy of a calculated result is limited by the least
accurate measurement involved in the calculation.
11.5 is rounded to 12,
(1) In addition and subtraction, the result is rounded
off to the last common digit occurring furthest to the
right in all components. For example,
100 (assume 3 significant figures) +
23.643 (5 significant figures) = 123.643,
which should be rounded to 124 (3 significant figures).
This rule means that if the digit to be dropped is 5

followed only by zeroes, the result is always rounded
to the even digit. The rationale is to avoid bias in
rounding: half of the time we round up, half the time
we round down.
D. Rules for counting
(2) In multiplication and division, the result should be Counted numbers have an infinite number of signifirounded off so as to have the same number of significant figures:
cant figures as in the component with the least number
of significant figures.
10 notebooks + 285 notebooks = 295 notebooks
For example,
3.0 (2 significant figures ) 12.60 (4
significant figures) = 37.8000
which should be rounded off to 38 (2 significant figures).
C. Rules for rounding off numbers

(1) If the digit to be dropped is greater than 5,
the last retained digit is increased by one. For
example,
(2) If the digit to be dropped is less than 5, the
Revised Fall 2014
This section on significant figures was taken in part from:

http://www.chem.sc.edu/faculty/morgan/resources/sigfigs/
index.html
For great practice see:
http://science.widener.edu/svb/tutorial/sigfigures.html
Page 10
Worksheet score: ____________

Laboratory #2 Worksheet
Date:
Name:
Section:
Female (g)
209
222
456
_________ m
256
_________ mm
185
_________ mm
I. The Metric System

1. Length
a) 2.4 cm
b) 264 cm
c) 23 m
=
=
=
Ratio:
2. Mass
a) 0.85 g
b) 5.3 g
c) 280 ng
Male (g)
225
198
356
235
456
=
=
=
_________ kg
_________ mg
_________ g
3. Volume
a) 53 L
=
b) 7.95 L
=
c) 0.058 mL =
_________ mL
_________ L
_________ L
4. Time
a) 0.120 sec = _________msec
b) 240 msec=
_________ sec
c) 0.059 msec= _________ nsec
5. Temperature
7. Your rats are to be fed a special diet in which each 70 g

portion of chow will contain
45 g of protein.
in 10 seconds (use your left index finger applied to the radial

artery of your right wrist).
Pulses per 10 seconds: _________
Calculate the number of pulses in
60 seconds (1 min.) using the
following ratio:
Pulses
10
? Pulses
60 sec
(or just multiply by 6)
a. How much protein would be Pulse rate (pulses/min): ________

present in 500 g of rat
9. On an EKG strip a nurse deterchow?
mines that a patients heart is
beating 5 times every 4.5 seconds.
b. What is the ratio of protein
to non-protein in the diet?
Calculate the time for each

heart beat: ______________
Calculate the patients heart rate
in beats/min: ____________
Hint: Divide 60 by the time per beat.
8. When you want to quickly

10. On the following page, graph
measure a persons pulse rate
the data set shown below:
you dont usually count the
number of pulses in a full miHeartRateDuringExercise
nute. You count the number of
II. Computations
pulses in 6, 10, or 30 seconds.
HeartRate
You
can
do
this
because
you
Time(min)
(beats/min)
6. You have 5 female rats and 5
know
that
if
you
set
up
a
proper
male rats, their masses are given
0
70
below. What is the mean ratio of ratio you can quickly and easily
calculate
the
pulse
rate
in
pulses
mass of the male to the female
3
78
per minute.
rats? Note: all ratios should be
simplified to x:1.
5
88
Measure your pulse rate by
counting the number of pulses
10
102
a) 72 F
b) 98.6 F
c) 2 C
Revised Fall 2014
=
=
=
_________ C
_________ C
_________ F
Page 11
Name:
(continued)
Revised Fall 2014
Page 12
Biology 335 Human Physiology: Transport
Laboratory #3
Cell Transport Mechanisms and Permeability
Objectives
simple diffusion through a nonliving semipermeable

membrane. This is called dialysis. We will also ex-
After completing the following simulation you should

firmly understand the concepts of simple diffusion,
facilitated diffusion, osmosis and active transport
across a cell membrane.
Starting the Program
Background
The fluids that bathe all of the cells of the body are
water-based (aqueous) solutions. Within any solution
the movement of the molecules dissolved within the
solution (solutes) is driven by the random collisions of
molecules in the solution. These collisions cause molecules which are collected together to be pushed apart.
This is diffusion. The movement of molecules from a
point of high concentration to a point of low concentration is because of random molecular motion.
If a semipermeable membrane (like a cell membrane)
blocks the movement of solutes (into or out of a cell)
but not the movement of the solvent, the solvent will
diffuse from a point of high solvent concentration (low
solute concentration) to a point of low solvent concentration (high solute concentration). This is called osmosis.
The movement of molecules across a cell membrane
can be passive (requiring no direct energy) or active
(requiring energy in the form of ATP). Passive
transport includes facilitated diffusion and filtration.
Facilitated diffusion is the movement of molecules
from a point of high concentration on one side of a
cell membrane to the other side through protein channels in the membrane. Filtration is the movement of
molecules, driven by hydrostatic pressure, through
protein channels (pores) across a biological membrane.
amine the diffusion of water across such a membrane

(osmosis) as well as studying the processes of filtration and active transport.
1. Insert the PhysioEx 9.0 CD-ROM into the CDROM drive of the computer or access the PhysioEx folder on the desktop.
2. If you started with the CD-ROM a browser window with the PhysioEx opening page should
open. If you started with a folder on the desktop
click on the StartHere icon .
3. Then click on Access PhysioEx 9.0 to start the
program.
4. Once the PhysioEx 9.0 windows opens click on
Exercise 1: Cell Transport Mechanisms and Permeability.
5. Beginning with the Overview, complete the Activities. At the end of each activity you are given
the option of saving your work in a .pdf file. Do
so, and when complete, submit the files to your
instructor (via email or hard copywhichever the
instructor prefers). Save the files with unique file
name such as:
Hallsec03pex-01-01
Hallsec03pex-01-02
Hallsec03pex-01-03
Hallsec03pex-01-04
Hallsec03pex-01-05
Make sure the filename includes your name, section number and the exercise (-01) and activity
number (-01, -02, etc.) that you are submitting.
Active transport processes involve protein pumps located in cell membranes which utilize energy released
by the hydrolysis of ATP. This energy is used to
move molecules from one side of a cell membrane to
the other from a point of low concentration to a point
of higher concentration.
In this computer simulation we are going to examine
Revised Fall 2014
Page 13
Hemodialysis
Artificial kidney machines have been developed that make use of dialysis to purify the blood of persons
whose kidneys have ceased to function. Known as hemodialysis, this procedure has saved the lives of
many persons suffering from renal failure. In such machines, blood is circulated on one side of a semipermeable membrane (often cellophane) while a special dialysis fluid is circulated on the other side. The dialysis fluid must be a solution that closely matches the chemical composition of the blood. Metabolic waste
products such as urea and creatinine diffuse through the membrane into the dialysis fluid and are discarded, while loss by diffusion of substances necessary to the body (such as sodium chloride) is prevented by
their presence in the dialysis fluid. From: http://www.answers.com/topic/dialysis
http://www.lrh.com.au/home/orginfo/departments/dialysis/1391image003.jpg
http://www.medicinenet.com/dialysis/article.htm#1whatis
Revised Fall 2014
Page 14
Biology 335 Human Physiology: Introduction to iWorx
Laboratory #4 Introduction to iWorx

Objectives
1.
2.
3.
Become familiar with the iWorx recording and

stimulating apparatus.
Examine the effects of exercise on pulse rate and
blood flow
Examine the effect of increasing stimulus frequency on threshold (summation).
Background
A stimulus is a change in the environment to which a
cell, organ or organism is sensitive. Irritable (or excitable) cells, such as neurons, muscle cells, and glandular cells, can respond to such a change if the stimulus
is of an appropriate type for a particular cell.
(Environmental changes can be thermal, chemical,
physical, electrical, etc.). For a cell to respond, the
stimulus must also be of sufficient magnitude (i.e., at
or above a threshold level of intensity). Some cells,
such as skeletal muscle cells, will respond to such
stimulation in a manner that can be readily observed.
Other cells, such as neurons, are responsive to stimuli,
but the response cannot be detected without sophisticated instrumentation.
Because of the relative ease of observing and recording contractions of skeletal muscle, we plan to use this
tissue in another laboratory as a means to explore
stimulus-response relationships. Specifically, we will
examine how the gastrocnemius muscle of a frog responds to various types of electrical stimulation. To
be successful in exploring these responses, it is important to learn how to use an electronic stimulator to
deliver carefully controlled electrical stimuli. The
parameters of electrical stimuli that physiologists often
manipulate are voltage (strength or intensity of stimulus), frequency of application (number of stimuli delivered per second) and duration of each individual
stimulus.
It is also important to record the responses to such
stimuli. The iWorx 214 data acquisition system receives electrical inputs via various electrodes and sensors which plug into the front panel of the iWorx 214.
Revised Fall 2014
This unit also has a built-in stimulator (red) which can

be used to mimic electrophysiological events in order
to record and analyze electrical responses. The basic
system is illustrated in Figure 1. This data collection
unit interfaces with the computer via the LabScribe
software which permits the display and analysis of a
wide variety of physiological responses including frog
skeletal muscle contraction, human heart rate, pulse,
lung volumes, etc.
Recordings of physiological events and their analysis
can be saved to the computer desktop and/or printed
from the computer.
Figure 1. iWorx 214 Front and Back.
Learning To Use LabScribe-The Basics

Turn on the iWorx 214 console using the power button
on the rear panel. On the computer, start the LabScribe software by double clicking on the icon located
on the desktop. After the software is loaded, select
Load Group from the Settings menu. When the dialog box appears, select 4 Intro to iWorx and then
click Load or Enter. Click on the Settings menu
again and select the Introduction to iWorx Recording settings file. Figure 2 shows the resulting screen.
Notice that each channel has its own (white) recording
area with a colored bar above it containing a title, Autoscale and add function select buttons, and the voltage value. Above the top channel colored bar is a time
value, the sampling speed, the display time, the Mark
comments, a T2-T1 value and the Record button.
Page 15

scriptions into the Marks area and
after hitting the Enter key the comment will appear on the recording.
The comments associated with a mark
can be moved vertically by clicking on
the comment and dragging it using the
mouse. Comments in a given view can
be reset to the lower margin by selecting Reset Marks under the View
menu.
Cursors
You can use the single cursor to

place comments or marks anywhere
in your recording. Just move the
cursor where you want the mark to
appear, type your comment into the
marks space and hit Enter.
Figure 2. LabScribe Introduction to iWorx screen
To demonstrate this:
You can also use marks to move through your recorded data. Pull down the Windows menu and select
Marks, or click the Marks icon on the toolbar. This
will bring up a list of all of the marks you have typed
into the record. Click on the time or comment for the
mark you wish to go to and then press Go To. You
can also delete marks from this menu if you inadvertently place a mark in the wrong position.
1.
Measurements
Screen Time
The default value for the time a signal crosses the
screen is 10 seconds. This display time can be
changed by clicking the display controls on the LabScribe toolbar.
2.
3.
4.
Click on Record then

after a few seconds click on Stop.
Click the left icon (big mountain) and notice that
the trace spreads out the display time is five
seconds.
Click the right icon (double mountains) twice
and see that the display time increases to 20 seconds.
Click the left mountain icon once to return to a
10-second display time.
Marks
The recording can be annotated by adding
marks in two ways:
1.
Measurements are made using the cursors. These are vertical blue lines that
span all channels and can be called up using one of the
cursor icons.
Using two cursors (left icon button) allows you to determine the difference in time or voltage between any
two points on the recording. Click and drag the lines
to the left or right to display the difference in:
Time (horizontal distance) is measured by determining the time between the positions of the cursors. This difference is labeled T2-T1 and is displayed at the top right next to the Record button.
When not recording, two blue vertical lines or

cursors will overlay the screen (Figure 2). These Voltage (vertical distance) is measured by determing the difference between the two small
cursors can be used to make measurements of
cross-shaped indicators on the cursor lines. This
your data. However, if you type a comment in the
difference is labeled V2-V1 and is shown in the
Marks area at the top of the screen using the keyarea on the upper right margin of each channel.
board and press the Enter key, the comment will
be entered in the lower margin at the left cursor
(after you hit the Record button).
2. While recording, you can type short deNote: If the iWorx 214 unit and computer are not
communicating you will receive an error. If this happens, click on the Tools menu then Find HardPage 16
Revised Fall 2014
ware. A small window should appear that says
Hardware Found=iWX114. Click OK and proceed.

Preparation and Using the Software
Journal
Although there is a built in journal feature in the Labscribe software, you should use Microsoft Word to record your data journal for all iWorx laboratories in this
class. To start Microsoft Word, click on Start on the
Windows task bar then click on the Word icon. Keep
Word open while you work and you can copy your Labscribe screen using Ctrl-C and paste it into Word using
Ctrl-V.
If necessary, turn on the iWorx 214 console using the

power button on the rear panel. On the computer, start
the LabScribe software by double clicking on the icon
located on the desktop. After the software is loaded,
select Load Group from the Settings menu. When the
dialog box appears, select 4 Intro to iWorx and then
click Load or Enter. Click on the Settings menu again
and select the Introduction to iWorx Recording settings file. Figure 2 shows the resulting screen.
In this exercise, the output from a pulse oximeter will be

used as a signal source which we will use to determine a
volunteers pulse rate and the relative rate of blood flow
through his or her finger/thumb before and after exerFor every exercise which utilizes the LabScribe software cise. We are testing the very simple hypothesis that
you should open a new journal and enter your name, lab exercise increases pulse rate and
blood flow. What do you think
partners names, exercise, lab section and date in the
will happen?
following format:
This document can be saved to a USB Flash Drive or
emailed to yourself or your instructor. Make sure you
save your data file and journal frequently.
Your Name
Lab Partners names
Exercise Number (Chapter in the manual)
Laboratory Section
Date
Recorder Procedure:
1.
2.
As you complete each exercise you can add headings

and insert your results directly into the journal. At the
end of the lab you should print it immediately to include
with your lab report or email it to your lab partners or
instructor.
Please note that at the end of each chapter there is a
Journal Format. You should insure that your journal contains the exact components listed.
Learning To Use LabScribe-Recorder
3.
4.
In this experiment we will be using a pulse oximeter

attached to the iWorx 214 and the LabScribe software to
test the hypothesis that exercise increases pulse rate in a
human volunteer. In the process you will learn how to
use the LabScribe recording software and the iWorx data
acquisition system.
Equipment Needed
iWorx 214 and computer with LabScribe installed.

Pulse Oximeter
5.
6.
Revised Fall 2014
Place your volunteers finger

tip into the pulse oximeter
according to the diagram on the device.
Mark your recording before you start by typing a
description in the blank space next to the mark
button then click on Mark. [NOTE: before you can
add any marks to your screen you first have to record at least one second of data. Quickly hit Record
then stop before trying to add marks.] This will
insert a mark on the screen wherever the left most
cursor is found. It will also be entered into an
index which will allow you to move to any mark
in your data as you proceed.
Click Record to record the finger pulse while at
complete rest.
Click Autoscale in the top channel (labeled Pulse)
title area and see the rhythmic signal almost fill the
channel recording area. Your data should look like
the example shown in Figure 3. If the signal is
upside-down you can right click with the mouse
on the window and invert the trace. If the baseline in the blood flow window isnt level you
should stop recording and then start it again. Repeat this process until you obtain a level baseline.
The software uses an algorithm to smooth the baseline whenever you hit Record. There are times
when this algorithm doesnt work very well and
you need to restart it.
After obtaining a good recording, click Stop to halt
recording.
To determine the heart rate, position the left cursor
Page 17

and post-exercise screens (2 each) should
be in the journal.
13. To save your recording, click on the
File menu and select Save As. Choose a
descriptive file name and save your recording to the desktop. Dont forget to also
save your journal.
14. Ask the other lab groups what their
results were. How did they compare to
your results?
Please note: When you proceed to the
next exercise you need to use a different
file name or you will lose your data for
this exercise! You can continue using the
same Microsoft Word document for
your journal.
Figure 3. Pulse and Blood Flow Data
on the peak of one of the pulse peaks in the top
channel. Position the right cursor on the fifth
peak following the first one you chose (see
Figure 4).
7. To Copy this screen to your journal switch
to your Microsoft Word journal file. Enter your
name and other relevant information to the top
of the journal page. To insert your data, press
the print screen key on your keyboard while in
Labscribe then switch to the word document
and hit Ctrl-V to paste it into the Word document.
8. The time for these 5 pulse cycles is listed as T2
-T1 in the upper right hand of the computer
screen right next to the Record button (see
Figure 4). Divide 300 by this number to get
the pulse rate (see Figure 5).
9. The height of the peaks (V2-V1) in the blood
flow window represents the blood flow through the
finger (ml/min). Your results may be a negative
number this is due to the arbitrary positioning of
the cursors when you measure the trough to peak
height. This isnt a problem; just take the absolute
value of the measured blood flow. We will be using
this procedure later in the course (see Figure 6).
Copy this screen to your journal as well.
9. Position the left cursor at the trough (bottom) of the
first full blood flow (lower window) peak and the
right cursor at the peak (top) of the same peak (see
Figure 6 on the next page).
10. Copy the screen to the journal.
11. Record the blood flow in the journal.
12. Repeat the above procedure after exercising for 3
minutes. Make sure that you add a mark before
you hit record every time! Both the pre-exercise
Revised Fall 2014
Figure 4. Determining Pulse Rate
Learning To Use LabScribe-Stimulus/

Response
Now that you are a little more familiar with the LabScribe software and what it can do, in this next exercise
you will be recording some characteristics of stimulus/
response relationships. The stimulus will be an artificially applied potential change which will change the
membrane potential of receptor cells in your tongue. If
the applied depolarization is above threshold for those
receptors, you will feel the stimulus as a slight electric
shock. The threshold voltage is characteristic of the
Page 18

cells which are being stimuCalculating Pulse Rate
the menu.
lated and how readily the
We are using 5 pulses to determine the pulse
applied potential change
Using the Stimulator
rate. If we divide the time for 5 pulses by 5 this
penetrates into the tissue. In will give us the time between each pulse. If we
addition, the frequency of
then divide 60 by that number we will be calcu- To bring up the stimulator panel (Figure 7)
lating the number of pulses in 60 seconds.
stimulation can also affect
click on the View menu then on Stimulator
These calculations can be simplified as shown
the threshold voltage bePanel.
below:
cause as you apply more
stimuli in a short period of
To Vary Voltage (strength of stimulus)
60
time the stimuli will sumTime for 5 pulses
mate. Thus, the threshold
The range of voltages which can be applied
5
for a single stimulus should
are 0 to 5 volts which is adjusted using the
be higher than the threshold
5
Amp (short for amplitudenot amps) win60 X
voltage when using 50 stimdow in the stimulator panel (Figure 7) in the
Time for 5 pulses
uli per second. This means
upper left hand corner of your computer
that many subthreshold
screen. Note: HP is the holding potential
stimuli can add together to
300
and should always be left at 0 for our purposPulses/minute es. W(ms) is the pulses width and should be
produce an above threshold
Time for 5 pulses
potential change in the tarat 5 or 10 ms.
get cells. This is referred to as
Figure 5. Calculating Rates
summation of subthreshold
To vary frequency of stimulation
stimuli. The iWorx 214 apparatus includes a built-in
stimulator which we will use in this exercise. We are
The frequency of stimulation can be adjusted in a similar
testing the hypothesis that increasing frequency of stimu- manner up to 100 per second using the F(Hz) (frequency
in stimuli per second or hertz) window.
=
=
To change the number of stimuli

The number of stimuli you apply can be adjusted by changing the #pulses from 1 up to 9999.
A 0 in this box forces the instrument to apply
continuous stimuli.
Note: Whenever you change any stimulator
parameter you need to click on Apply before
hitting start. When you click the Record
button the stimulator will automatically apply
the stimulus you have defined. If you click on
Apply after you hit record, the stimulator will
apply a new set of stimuli every time you click
on the button.
Setup:
Figure 6. Determining Blood Flow
lation
will reduce the threshold voltage needed to detect a stimulus
applied to the human tongue.
Set up the Software
Click on Settings, then Load Group, and Introduction to iWorx Stimulus. Click on Settings again then
on Introduction to iWorx Stimulus at the bottom of
The stimulating electrode should be connected to the red

and green terminals of the red stimulator panel on the
iWorx 214. On the LabScribe software screen, click on
Settings and choose Introduction to iWorx Stimulus.
You should now see two screens, the lower recording
screen is titled Stimulus while the upper is titled Response.
Figure 7. Stimulator Panel

Revised Fall 2014
Page 19

Find the event marker (Figure 8) and, if necessary, insert the DIN cable into the Channel 4 connector of the
IWorx 214 unit. Whenever the human volunteer presses the event button, this will produce a square wave
deflection on the top screen (Response) while the stimulus will appear in the bottom screen (Stimulus; see
figure 9). The stimulus is automatically displayed as a
spike indicating every time the stimulator applies current to the electrodes.
2.
lus frequency (summation of subthreshold stimuli)

on threshold, repeat steps 3 to 7 in the above exercise after increasing the frequency to 50.
Copy these data into the journal.
Amp=5.0 V
W(ms)=5
F(Hz)=1
#pulses=5
Skin response.
The skin of your forearm has a thick cornified

layer of dead cells on its surface and is superficially dry. Do you think that these characteristics might alter the ability of an artificially applied voltage to stimulate the nerve cells found underneath the
epithelium?
Stimulus/Response Procedure:
Amp=Varies
W(ms)=5
1. Locate the stimulating electrode and genF(Hz)=1
tly clean it with an alcohol swab.
#pulses=0
2. To provide the volunteer with
some idea of the feeling of an above
threshold stimulus, set the Amp to 5
volts.
3. Locate the event marker (Figure 8) and
give it to your volunteer to hold in his or
her left hand.
4. Gently place the stimulating electrodes
on the surface of the volunteers tongue.
Figure 8. Event
5. Add a mark indicating the stimulus paMarker
rameters as shown in the small box
above. Click on Mark.
6. Click on Record. The volunteer should readily
feel the resulting stimulus and press the event
marker button accordingly. Click on Stop.
7. To determine the threshold voltage for the same
response, reset the voltage to 0.1 V (Amp, make
sure you click on Apply) and press Record again.
Whenever the volunteer feels the electrical stimulus they should press the button firmly and hold it
down as long as they feel the stimulus then release
it. After the five stimuli are applied click on Stop.
If the volunteer cannot feel the stimulus they
should not press the button.
8. Increase the voltage in 0.1 or 0.2 V units, click on
Apply, enter the stimulus voltage as a Mark then
click on Record again. Repeat this process until
the volunteer can feel the stimulus.
9. The magnitude of the weakest stimulus the volunteer can feel represents the threshold for a single
stimulus.
10. Copy a recording of the threshold voltage data into
the journal. Save your file using a different
name than you used for the previous exercise.
1. At the maximal voltage (5 V) see if your

volunteer can detect the stimulus on his/her
forearm.
2. If your volunteer could detect a skin response place this recording in the journal.
Save your journal.
Response
Stimulus
Figure 9. Stimulus Response
Amp=Varies Effects of increased stimulus frequency on

W(ms)=5
threshold (summation).
F(Hz)=50
#pulses=0
1. To examine the effects of increased stimuRevised Fall 2014
Page 20
Journal: __________
Worksheet Total: ___________

Date:
Name:
Section:
Learning to use LabScribe - Recorder

1) What was your volunteers pulse rate (pulses per minute) before and after exercise as determined using the
pulse oximeter?
_Before:___________________________________ _After:_________________________________________
2) How does the pulse rate that you recorded relate to that persons heart rate?
___________________________________________________________________________________________
3) What was blood flow through your volunteers finger before and after exercise?
_Before:___________________________________ _After:_________________________________________
4) If the blood flow changed, what do you think the physiological significance of that change is?
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
5) If you change the display time in the main window, will this change the pulse rate that you measured? (Try it)
Explain your answer.
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
6) What effect did exercise have on the pulse rate and blood flow of your volunteer? Did your results support the
hypothesis mentioned in the exercise?
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
Revised Fall 2014
Page 21
Name:
Laboratory #4 Worksheet (continued)

(cont.) Name: _______________
Learning to use LabScribeStimulus/Response
7) Enter the tongue threshold voltages for the classs volunteers in the table below. Calculate the class average
thresholds at 1 and 50 stimuli per second (Hz).
Tongue Threshold Voltages Class Data

Student:
F(Hz) = 1
F(Hz) = 50
#1
#2
#3
#4
#5
#6
Averages:
8) You should have seen a difference between the threshold voltage at 1 stimulus per second compared to 50 stimuli per second. Why should the threshold voltage change with a change in stimulus frequency? Did your data
support the hypothesis stated in the exercise?
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
9) You should see that there was a delay between the administration of a stimulus to the tongue and the volunteers
response to that stimulus. What caused this delay?
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
10) Did your volunteer feel the stimulus on his or her forearm? Explain these results.
___________________________________________________________________________________________
Revised Fall 2014
Page 22
Name:
Laboratory #4 Worksheet (continued)

(cont.) Name: _______________
11) Explain the process whereby an electrical stimulus can induce a receptor cell to generate action potentials.
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
Note: Make sure that you hand in your journal

for all exercises with this worksheet.
Revised Fall 2014
Page 23
Journal Format for Introduction to iWorx
(cont.)
(Guide for producing
Your Name
Lab Partners names
Learning to Use Labscribe
Laboratory Section
Date
Name: journal)
_______________
a complete
Every Journal should include every component listed in this journal format guide. Hand
the complete journal in with your worksheet!
A. RECORDER PROCEDURE
Pulse rate and blood flow before exercise
Paste the screen showing the pulse rate
measurement of the volunteer from
your lab group.
State how the pulse rate was determined.
Time for 5 recorded pulses = ______________
300/time for 5 recorded pulses = ______________ = the pulse rate in pulses/minute
Paste the screen showing the blood
flow measurements of the volunteer
from your lab group.
State the blood flow.
Blood flow = ______________ ml/minute
Pulse rate and blood flow after exercise

Paste the screen showing the pulse rate
measurement of the volunteer from
your lab group.
State how the pulse rate was determined.
Time for 5 recorded pulses = ______________
300/time for 5 recorded pulses = ______________ = the pulse rate in pulses/minute
Paste the screen showing the blood
flow measurements of the volunteer
from your lab group.
State the blood flow.
Blood flow = ______________ ml/minute
Revised Fall 2014
Page 24
Journal Format for Introduction to iWorx (continued)
(cont.)
(guide for
setting
_______________
up Name:
your journal)
B. STIMULUS/RESPONSE DATA
Tongue threshold voltage
Paste the screen showing the sub threshold and threshold voltages and response
of your volunteer.
State the threshold voltage .
Threshold voltage (Amp) = _________________ volts
Frequency of stimuli (should be 1) = _______________ Hz (stimuli/second)
Tongue threshold voltage with increased frequency of stimulation
Paste the screen showing the threshold
voltage with increased frequency of
stimulation and the response of your
volunteer.
State the threshold voltage.

Threshold voltage (Amp) = _________________ volts
Frequency of stimuli (should be 50) = _______________ Hz (stimuli/second)
Skin response
Paste the screen showing the stimulus
and response of your volunteer (only if
there is a response).
State the voltage .
Amp = 5V
State the frequency of stimulation
F(Hz) = ___________________
Make sure all 7 journal pages are in order and turn them in with the worksheet.
Revised Fall 2014
Page 25
Revised Fall 2014
Page 26
Biology 335 Human Physiology: Frog Muscle
Laboratory #5
Physiology of Skeletal Muscle
laboratory. If our depolarization is above the threshold
value for the myofibers then the muscle will experience an action potential and the result will be a muscle
1. To stimulate and observe skeletal muscle contrac- contraction.
tion.
2. To understand the properties of a twitch, summa- The gastrocnemius muscle of a frog will be used as
tion of contraction and recruitment.
your test muscle. Dissection of the muscle from the
leg of the animal damages its normal source of stimulation the sciatic nerve. Direct stimulation of the
Background
muscle with the stimulator allows us to control various
aspects of the stimulation received by the muscle.
Remember that you can vary the intensity (strength)
In the human body, as in the bodies of frogs and other of the stimulus delivered to the muscle by adjusting
vertebrate animals, skeletal muscle function is directed the voltage; you can also vary the frequency of stimuby the somatic division of the nervous system. Direct lation by adjusting the number of stimuli delivered per
nervous stimulation is normally required for a skeletal second.
muscle to contract; in the absence of such stimulation
skeletal muscles remain in a relaxed state. In this laBy altering the strength of the stimulus you will be
boratory, you will examine how such a muscle rechanging the number of muscle fibers involved in the
sponds to stimulation. More specifically, you will
resulting contraction. In the intact animal recruitment
focus on how difof motor units is utilized to vary the strength of the
ferent patterns of
muscle contraction. We will be using different voltstimulation result
ages to mimic this process in the isolated muscle.
in different types
of muscle activity.
Changes in stimulus frequency can also alter the
strength of contraction. However, in the intact aniRemember that
mal, contraction is induced by a chain of stimuli
the generation of
which are above the fusion frequency for the musan action potential
cle. The fusion frequency is the frequency of stimuin an excitable
lation which produces a completely smooth, tetanic
tissue like skeletal
contraction of the muscle. It results from the summuscle first remation of overlapping muscle contractions and the
quires that the
accumulation of calcium within the sarcomeres.
membrane be
We can determine the fusion frequency of the frog
brought to threshgastrocnemius muscle by varying the frequency of
old. In skeletal
stimulation which we apply.
muscle this occurs
normally at the
Preparation
neuromuscular
junction under the
influence of acetylcholine reFigure 1 illustrates how the gastrocnemius muscle
leased from the alpha Figure 1. Gastrocnemius muscle
will be positioned within the experimental equipment
prepared for recording.
motor neuron. In
as you assemble it for todays lab. You will prepare
todays laboratory
the muscle such that it remains attached to the frogs
we will be bypassing the neural stimulation by directly femur bone. The femur will be firmly held in place by
applying a membrane depolarization to the myofibers a femur clamp. This will stabilize the origin of the
using the same stimulators we used in the previous
muscle. The insertion of the muscle will be attached
Objectives
Revised Fall 2014
Page 27

trocnemius muscle will be dissected.
1. Grasp the cut end of the femur bone with forceps
and pull the skin off the leg.
2. Identify the femur, tibio-fibula (the two bones are
fused in the frog) and the gastrocnemius muscle
with its Achilles tendon.
3. Cut the quadriceps and hamstring muscles away
from the femur.
4. Insert a glass probe (see Figure 3) between the
gastrocnemius muscle and the tibio-fibula and
free it from the connective tissue by sliding the
glass rod up and down along the bone.
5. Tightly tie a piece of doubled thread approximately 30 cm long to the Achilles tendon.
Please note that in this exercise
Figure 2. Motion
6. Cut the Achilles tendon from the tibio-fibula disthe mass of the rod is a preload
Transducer
tal to the knot.
7. Cut the tibio-fibula
which acts to pull the
just below the knee (see
cytoskeletal elements
Figure 4).
within the sarcomeres
8. Position the femur in
into alignment prior to
Femur
Tibia/Fibula
the femur clamp as
contraction. In intact
shown in Figure 1. Place
organisms, muscle
slight tension on the
tone provides a premuscle by moving the
load which serves to
Gastronemius
femur clamp up until you
pull the cytoskeletal
can see that you are pullelements in the sarcoAchilles Tendon
ing up on the transducer.
meres of our muscles
9. Insert the two stimuinto alignment. This
lator wires through the
is important because it
muscle by firmly holding
decreases the time it
the muscle with one hand
takes for our muscles
Figure 3. The Frog Leg
while stabbing the wire through the
to respond to stimuli (latency period).
muscle with the other. The wires should be poFemur
sitioned as shown in Figure 1.
If the force generated by a muscle
contraction equals the preload , the
At this point you
muscle will shorten and lift the load
(mass actually moved). This type of
should consult your
contraction is referred to as an isotoninstructor to be sure
ic contraction because the forced
that your apparatus is
needed to move the load is constant
adjusted to provide
during the contraction. If the force
Figure 4.
optimal recordings of
generated by a muscle contraction is
muscle activity.
NOT enough to lift the load then the muscle does not
shorten. Since the length of the muscle stays the same
It is important to keep the muscle moist during the
it is referred to as an isometric contraction. In this
dissection and throughout the experimental procelaboratory, we are recording isotonic skeletal muscle
dures. Saline solution is provided in squeeze bottles
contractions.
for this purpose. This fluid must be used instead of
water, as its osmolality has been adjusted to prevent
A. Dissection of the muscle
osmotic damage to the muscle tissue. You should also
The anesthetized frogs will be killed by the instructor note that without an intact circulatory system or mechin a manner that has been approved by the RIC Institu- anism for replenishing nutrients and removing wastes,
tional Animal Care and Use Committee. Each group the isolated gastrocnemius preparation we are using
has a limited period of usefulness.
of students will receive one leg from which the gasto the motion transducer (Figure
2) via a thread tied to the Achilles
(calcaneal) tendon. After appropriate adjustments, contraction of
the muscle will result in movement of the transducer rod. Movement of the rod will produce a
permanent record on your computer that will provide you with a
information about how the muscle
has responded to different patterns
of stimulation.
Revised Fall 2014
Page 28
Figure 5. Stimulator
4.
Exercises
You should be thoroughly versed in all aspects of
the following exercises before beginning, as the
muscle will probably not last longer than about 30
minutes.B. iWorx Setup
After turning on the iWorx unit and starting up the
LabScribe software, click on Settings, Load Group
and select 5 Muscle Physiology. Click on Settings
again then choose 5 Muscle Contraction from the
drop down list. This will reveal a window containing
two channels. The top channel (Muscle Contraction)
will be used to record the muscle contractions while
the bottom channel (Stimulus) is reserved for recording the stimulator output .
5.
6.
7.
2. Click on Record then Stop to establish a record so you can begin adding marks or edit your
journal.
3. Add a mark to the recording indicating the
voltage.
One partner should watch the muscle while another
partner clicks on Record. As soon as you hit Record, 5 stimuli will be applied, however you can
apply additional stimuli by clicking on the Apply
button.
After a few stimuli click on Stop.
Increase the voltage by changing the Amp= to 0.2,
click on Apply then repeat steps 3 through 5.
Repeat steps 3-5, increasing the Amp each time
until you can observe the muscle twitch in response
to the stimulus. (See Figure 6).
It is important for someone to watch for the muscles

response and insure that you are recording properly.
8.
When the muscle finally responds, stop recording

then click on the double mountain display time icon
until voltages lower than threshold and the threshold
voltage data are both visible in one screen. Copy
these data to the journal. Remember to put your
heading in your journal (as described in the
Introduction to iWorx laboratory).
Amp=Varies A. Determination of Threshold

W(ms)=10
Remember that when you determine any
F(Hz)=1
threshold, you should start with very low inten#pulses=5
sity stimulation and work up to higher intensiB. Summation of Sub-threshold Stimuli
ties. You will be using the iWorx stimulator
Amp=Varies
panel as illustrated in Figure 5. Remember that if this
W(ms)=10
In this exercise you will determine whether the
panel is not visible, click on View and then Stimulathreshold voltage is different if the frequency of F(Hz)=50
tor panel.
stimulation is increased. Just as excitatory post #pulses=10
synaptic potentials (EPSPs) can summate at a
Start with the amplitude set at 0.1 Volts, W = 10, F
neurons axon hillock to bring a neuron to threshold and
(Hz)=1 and #pulses=5.
generate an action potential, below threshold
1. Whenever you change the settings in the stimula- (subthreshold) stimuli (artificially applied or natural) can
tor control panel hit the Apply button before hit- summate within the motor end plate of a skeletal muscle
fiber. Thus, many subthreshold stimuli might summate
ting Record or adding any marks.
within our isolated gastrocnemius muscle effectively
decreasing the voltage needed to stimulate muscle contraction and decreasing the threshold voltage.
Notice that the subthreshold
stimuli do NOT result in muscle
contraction, while the threshold
stimuli clearly produce muscle
contractions.
Subthreshold voltage
Set the F(Hz) at 50 stimuli per second and #pulses=10,

and repeat the determination of threshold (steps A1-A7),
starting again from the lowest possible voltage. Record
the data just as before and paste them into your journal.
Threshold voltage
C. Effects of Increasing Stimulus Intensity

(above threshold)
Amp=Varies
W(ms)=10
F(Hz)=1
In this exercise you will be stimulating the mus- #pulses=5
cle using increasing voltages to recruit more
Figure 6. Threshold stimulus

Revised Fall 2014
Page 29

D. Effects of Increasing Stimulus Frequency
As the frequency of stimulation increases you
will observe incomplete summation, treppe and
tetanus (complete summation). The following
exercises are designed to illustrate these processes.
Notice that the peak

height does not increase when you
increase the voltage
above the maximal
stimulus.
1.
Maximal stimulus
Figure 7. Maximal stimulus

myofibers into the contraction.
1.
2.
3.
4.
5.
2.
Return the frequency (F(Hz)) setting on the stimulator to 1 stimulus per second, set the Amp to the
threshold voltage (determined in exercise A) and
set #pulses=5.
Add a mark indicating the starting voltage.
Turn the stimulator and recorder on (click Record
in the LabScribe software).
After a few contractions, click the Stop button.
Increase the voltage in 0.2 volt increments and repeat steps 2 through 4 until the magnitude of the
contractions no longer increases. Remember to
click on Apply each time you change the voltage
and before you add your mark or hit Record.
3.
4.
Amp=Part C
W(ms)=10
F(Hz)=Varies
#pulses=0
Hold the intensity of stimulation constant at the

maximal voltage (if your muscle is already exhibiting signs of fatigue you might have to increase the
voltage to a level higher than the maximal stimulus). Set the frequency back to 1 per second but
now set the #pulses to 0 to force the stimulator to
apply constant stimuli at the defined amplitude and
frequency.
Click on Record in the software and you should see
the muscle twitch in response to the stimuli. Click
on Stop.
Increase the frequency to 2 and repeat step 2.
Repeat these steps increasing the frequency each
time until you obtain complete summation. Please
remember to mark your recording for each frequency.
As the frequency was increased to somewhere between 3

-6 stimuli per second you should have seen the muscle
contract in a pulsatile manner where a new contraction is
initiated before the previous contraction has fully relaxed. This is incomplete summation and should produce a staircase-like pattern of contraction on the recording which can be referred to as treppe (Figure 8). As
The lowest voltage which can produce the largest mag- you continued to increase the frequency of stimulation
nitude contraction is called the maximal stimulus (see you will eventually reach the fusion frequency. At this
Figure 7). Adjust the display time so that you can clear- point you will observe a smooth, complete contraction.
ly see the effects of increasing voltage and the maximal At this point your muscle is experiencing complete sumstimulus and copy this to the journal.
mation (tetanus).
E. Fatigue
Tetanus
Treppe
Amp=Part C
W(ms)=10
F(Hz)=50
#pulses=0
Muscle contraction is a very complex cellular

event which depends on the ionic and pH environment of the myofibers as well as a ready
supply of ATP to support cross bridge cycling.
Fatigue occurs progressively as the ionic and pH environment is disrupted, and the supply of ATP is depleted.
Without a blood supply which can replenish ATP stores
and maintain the ion and pH levels in the muscle tissue,
the isolated gastrocnemius muscle is more susceptible to
fatigue. In this exercise we are going to maximally
stimulate the isolated muscle and record the decreasing
contraction strength that occurs as the muscle fatigues.
Figure 8. Treppe and Tetanus

Revised Fall 2014
Page 30

called the contraction period and is measured as the time
from the onset of contraction to the peak of the contraction. The relaxation of the muscle takes a longer period
of time because it is due to the passive recoil of the series elastic elements within the muscle. This relaxation
period is measured from the peak of the contraction to
the point when the muscle has returned to its baseline
length.
Figure 9. Fatigue
Since this will effectively use up your isolated muscle, make sure that you have good data for all other exercises before trying this exercise.
1.
2.
3.
Use twitches
recorded in
Part C.
Set the stimulator to administer the maximal stimulus at a frequency of 50 stimuli/sec with #pulses set
to 0 or more. #pulses=0 provides continuous stimulation.
Click on Record.
Continue stimulating the muscle at a maximum
frequency until you see evidence of fatigue (Figure
9). Once you observe the contraction strength in
the Response Window decreasing, click on Stop,
adjust the display time appropriately and copy this
information to the journal.
Using data that you generated early in the laboratory

(Part C will work well), bracket a few good twitches
with the double cursors in the LabScribe software. Using the cursors measure the latency period, contraction
time, relaxation time (as shown in Figure 10) and total
twitch time (C+R). Repeat the measurements for 5 different twitches and place this data in the table in the
worksheet. Calculate the mean latency period, contraction time, relaxation time and total twitch time for your
muscle. Copy a representative example of each of these
screens into your journal (see Figure 10).
Make sure that you save your data file and print or
email copies of the journal for each lab member before leaving the laboratory. All calculations and
measurements should be clearly indicated in the
journal!
F. Properties of a Twitch
Although muscle twitches are not examples of normal physiologically meaningful

skeletal muscle contraction, they do provide some insight into the actual workings of
skeletal muscles. For each above-threshold stimulus, the skeletal muscle will exhibit a brief delay
before it exhibits any shortening (contraction).
This latency period is the time lag between the
onset of the stimulus and the onset of the resulting muscle contraction. This lag time is due to
the loose nature of the muscle tissue requiring
some initial muscle contraction before any shortening of the muscle is evident, as well as the time
required for the excitatory stimuli to induce muscle contraction. Once the muscle begins contracting, the contraction occurs fairly rapidly because
of the active cross bridge cycling induced by your
stimulus. This rapid period of contraction is
Latent
period
Contraction
period
Relaxation
period
Figure 10. Properties of a twitch

Revised Fall 2014
Page 31
Revised Fall 2014
Page 32
Journal: __________

Date:
Name:
Name:
Section:
1) What was the threshold voltage needed to stimulate contraction of the frogs gastrocnemius muscle when the
frequency was set at 1 stimulus per second?
____________
2) What was the threshold voltage necessary when the frequency was set at 50 stimuli per second?
____________
3) Did the threshold voltage change when you increased the stimulus frequency? How and why?
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
4) Describe the effect of increasing the intensity (voltage) of the stimulus while keeping the frequency constant?
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
5) Define maximal stimulus.
___________________________________________________________________________________________
___________________________________________________________________________________________
6) What is a motor unit?
___________________________________________________________________________________________
___________________________________________________________________________________________
7) Describe the mechanism by which intact human skeletal muscles exhibit graded contractions.
___________________________________________________________________________________________
___________________________________________________________________________________________
8) Are motor units involved in producing your data with the isolated gastrocnemius muscle? Explain.
___________________________________________________________________________________________
___________________________________________________________________________________________
Revised Fall 2014
Page 33
Laboratory
Worksheet
Laboratory#5#5
Worksheet (cont)
(continued)
Name:
Name: _______________
Name:
9) Explain how summation of sub-threshold stimuli works at the membrane/ion level.

___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
Data Table 1 Properties of a Twitch
10) Define a muscle twitch:
Twitch Example
_____________________________________________
_____________________________________________
_____________________________________________
Latency
Period
Contraction Period
Relaxation
Period
1
2
3
_____________________________________________
11) What is happening inside the myofiber during the latency period?
5
Mean
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
12) What is happening inside the sarcomere during the contraction period?
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
13) What is happening inside the sarcomere during the relaxation Period?
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
14) Why does summation of contraction occur? (Do not confuse this with summation of subthreshold stimuli!)
___________________________________________________________________________________________
___________________________________________________________________________________________
Revised Fall 2014
Page 34
Total
Twitch
Time
Laboratory
Worksheet
Laboratory#5#5
Worksheet (cont)
(continued)
Name:
Name: _______________
Name:
15) What was the fusion frequency for your isolated gastrocnemius muscle? What is the ionic basis for the fusion frequency?
Fusion Frequency: _____________
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
16) Why does muscle fatigue occur?
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
17) Since sarcomeres within skeletal muscles are rigidly aligned with each other what do you think excessive
stretch or compression (remember the basic structure of the sarcomere with overlapping thin and thick filaments)
will do to the force generation of a muscle contraction?
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
19) How does preload in an isolated muscle preparation relate to muscle tone in an intact organism?
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
Revised Fall 2014
Page 35
Journal Format for Physiology of Skeletal Muscle
(cont.)
Your Name
Lab Partners names
Laboratory Section
Date
Name: journal)
_______________
a complete
A. DETERMINATION OF THRESHOLD
voltage and resulting recorded muscle
contraction with F(Hz)=1
Threshold voltage at 1 Hz = ______________

B. SUMMATION OF SUB-THRESHOLD STIMULI
voltage and resulting recorded muscle
contraction with F(Hz)=50
Threshold voltage at 50 Hz = ______________

C. EFFECTS OF INCREASING VOLTAGE (ABOVE THRESHOLD)
Paste the screen showing before and
after maximal stimulus was reached.
Maximal stimulus voltage = ______________

D. EFFECTS OF INCREASING STIMULUS FREQUENCY
Paste the screen showing treppe
(summation of contraction) and tetanus.
Fusion frequency = _____________
Revised Fall 2014
Page 36
Journal Format for Physiology of Skeletal Muscle (continued)
(cont.)
(guide for
setting
_______________
up Name:
your journal)
E. FATIGUE
Paste the screen showing fatigue
What voltage and frequency did you use to induce fatigue? ______________
F. PROPERTIES OF A TWITCH
Paste the screen showing the measurement of latency period
Latency period = ______________
Paste the screen showing the measurement of contraction time.
Contraction time = ______________
Paste the screen showing the measurement of relaxation time.
Relaxation time = _____________

Revised Fall 2014
Page 37
Revised Fall 2014
Page 38
Biology 335 Human Physiology: Muscle Simulation
Laboratory #6
Skeletal Muscle
Physiology: Computer Simulation
twitch. A tracing of a muscle twitch is divided into
three phases: latency, contraction, and relaxation. The
latency phase (or latency period) is a short period beThe following exercises will explore some basic prop- tween the time of stimulation and the beginning of
erties of skeletal muscle contraction using a computer contraction. Although no force is generated during
simulation.
this interval, chemical changes occur intracellularly in
preparation for contraction (excitation contraction
coupling). During contraction, the myofilaments are
Background
sliding past each other and the muscle shortens. Relaxation takes place when contraction has ended and the
muscle returns to its normal resting state and length.
Skeletal muscles are composed of hundreds to thousands of individual cells, each doing their share of
Starting the Program
work in the production of force. As their name suggests, skeletal muscles move the skeleton. Skeletal
muscles are remarkable machines; while allowing us
1. Insert the PhysioEx 9.0 CD-ROM into the CDthe manual dexterity to create magnificent works of
ROM drive of the computer or access the Physioart, they are also capable of generating the brute force
Ex folder on the desktop.
needed to lift a 45 kg (~100 lb) sack of concrete.
2. If you started with the CD-ROM a browser window with the PhysioEx opening page should
When a skeletal muscle from an experimental animal
is electrically stimulated, it behaves in the same way
as a stimulated muscle in the intact body, that is, in
3. Then click on Access PhysioEx 9.0 to start the
vivo. Hence, such an experiment gives us valuable
program.
insight into muscle behavior.
4. Once the PhysioEx 9.0 windows opens click on
Exercise 2: Skeletal Muscle Physiology.
A contracting skeletal muscle will produce force and/ 5. Beginning with the Overview, complete the Acor shortening when nervous or electrical stimulation is
tivities. At the end of each activity you are given
applied. Unlike single cells or motor units, which
follow the all-or-none law of muscle physiology, a
so, and submit to your instructor. Save the files
whole muscle responds to stimuli with a graded rewith unique file name such as:
sponse. A motor unit consists of a motor neuron and
all the muscle cells it innervates. Hence, activation of
Hallsec03pex-02-01
the neuron innervating a single motor unit will cause
Hallsec03pex-02-02
all muscle cells in that unit to fire simultaneously in an
Hallsec03pex-02-03
all-or-none fashion. The graded contractile response of
Etc.
a whole muscle reflects the number of motor units
firing at a given time. Strong muscle contraction imMake sure the filename includes your name, secplies that many motor units are activated and each unit
tion number and the exercise (-02) and activity
has maximally contracted. Weak contraction means
that few motor units are active; however, the activated
units are maximally contracted. By increasing the
number of motor units firing, we can produce a steady
increase in muscle force, a process called recruitment
or motor unit summation.
Objectives
Regardless of the number of motor units activated, a

single contraction of skeletal muscle is called a muscle
Revised Fall 2014
Page 39
Revised Fall 2014
Page 40
Biology 335 Human Physiology: Reflexes
Laboratory #7
Reflexes
Objectives
1.
2.
3.
To observe electromyography activity associated

with muscle contraction.
To observe the Achilles tendon and patellar reflexes and measure the conduction velocity of the reflex arc.
To observe the effect of conscious motor activity
(Jendrassiks maneuver) on the reflex arc.
stretching the receptors. These fibers are innervated

by gamma motor neurons. The majority of a muscle
consists of extrafusal muscle fibers, which are innervated by alpha motor neurons and are responsible for
muscle shortening and production of muscle tension.
Background
Skeletal muscles have specialized receptors which convey information about muscle length, tension, and pressure to the central nervous system. The sensory receptors responsible for providing information about the
length, or the rate of change of the length, of a muscle
Figure 2. Myotactic reflex

When a muscle is stretched, excitation of its spindle
fibers causes a reflexive contraction of the muscle (see
figure 2). This reflex response is known as a stretch
(myotactic) reflex. The minimal delay between the
muscle stretching and the reflex contraction is due to
its monosynaptic pathway. The sensory afferent
nerves from the spindles synapse directly with alpha
motor neurons:(there are no interneurons in the pathway). This pathway is the simplest possible reflex arc.
Figure 1. Muscle Spindle
As an example of the stretch reflex, consider the reflex

are called muscle spindles (containing spindle fibers or response that occurs when a person jumps from a low
intrafusal muscle fibers, see figure 1). Arranged in par- stool to the floor. The extensor muscles of the legs are
allel with the contractile muscle fibers (extrafusal mus- stretched on landing, lengthening all their muscle spincle fibers), the spindles are stretched when the muscle is dles. The discharge of the muscle spindles is constretched by an external force. Therefore, these recepveyed to the central nervous system through the fasttors play a significant role in reflexes and maintaining
conducting alpha afferent axons. These sensory axons
muscle tone. Muscle spindles contain a small bundle of enter the spinal cord through the dorsal root and synintrafusal muscle fibers which do
apse with the motor neurons of
not contribute to muscle shortening This laboratory exercise is modified
the same extensor muscle. In
or force production, but regulate the from: iWorx Physiology Laboratory
turn, the motor neurons trigger
excitability of the sensory afferent
the contraction of the extensor
Manual, Exercise HN-2.
spindle nerves by mechanically
muscle to oppose the stretch
Revised Fall 2014
Page 41

produced by landing, completing the reflex arc. This
reflex is one of the main reasons you keep your balance
and do not fall when changing certain body positions.
You will be recording electromyograms (EMGs), the
summation of asynchronous electrical activity (muscle
action potentials) in the multiple fibers in the muscle,
and use them to determine the time between the stretch
of the tendon and the arrival of the motor impulse at the
muscle.
B. iWorx Setup
After turning on the iWorx unit and starting up the LabScribe software, click on Settings, Load Group and
select 7 Reflexes. Click on Settings again then choose
Reflexes from the drop down list. This will reveal a
window containing two channels. The top channel
(EMG) will be used to record the EMG activity while
the bottom channel (Tendon Tap) is reserved for recording the tap on the tendon.
Preparation for Achilles Tendon

A. Electrode Placement
1. Use an alcohol prep pad to clean and abrade
three regions on the calf of the left leg for electrode attachment. One area is near the ankle,
the second is on the skin near the center of the
gastrocnemius muscle (calf) and the third is
about 6 cm below the back of the knee. Let the
areas dry.
2. Place a disposable electrode onto each of the
areas.
3. Attach the red (+1) lead to the electrode near
the back of the knee.
4. Attach the black (-1) lead to the electrode in
the middle of the calf.
Figure 4. Achilles Tendon Reflex
5. Attach
the green
(C ) lead to
the electrode Exercises
near the ankle.
A. Achilles Tendon Reflex
Figure 3. Achilles Tendon

reflex electrode placement.
Conduction Velocity ( m/sec) =
A volunteer in each lab group should sit with their leg

swinging freely off the ground (you can raise the stool if
you need too). The Achilles tendon connects the gastrocnemius muscle to the tarsal bone of the foot. When
you tap on this tendon it will stretch the gastrocnemius
muscle and activate the spindle fibers of that muscle.
The reflexive response to such stretch produces contraction of the gastrocnemius and a downward movement of
the foot (plantar flexion).
Click on Record and have the volunteer rapidly flex
his foot once, pause and repeat. You should now be
recording the EMG activity (Figure 5) associated with
the contraction
Total Path Length ( mm )

( M ean Reflex Time ( msec) - 0.5 msec )
Figure 5. EMG recording

Revised Fall 2014
Page 42

of the gastrocnemius muscle. After clicking on
Stop, copy this recording (using CTRL-C) to
your journal.
Response
Mark the recording with the subjects name and

Achilles Tendon Reflex then hit Record.
With the volunteer completely relaxed tap the
Achilles tendon with the reflex hammer. Record 5
good examples of the Achilles reflex response.
Click Stop after the 5th trial. Paste one good
example of the reflex recording in your journal.
Measure the time between the first peak of the
tendon tap and the onset of the EMG spike. Then
measure the greatest peak to trough magnitude for
the EMG spike for each of the five trials and record these numbers in the worksheet. Calculate the
mean Achilles reflex response time and the mean
magnitude. Measure the distance between the
black electrode and the L5-S1 level of the spinal
cord (on the back at the level of the top of the hips).
Remember to double this distance to determine the
total path length. Calculate the conduction velocity
using the formula in the box at the bottom of the previous page.
Record the data in the worksheet (Data Table 1).
Tap
Figure 6. Recording Reflex Time and Magnitude
the green (C ) lead should be placed on the side of the

knee (this is the ground).
Click on Record and have the volunteer extend their
leg rapidly once. Repeat this a couple of times then
click on Stop. Copy the screen shot of the EMG
recording in your journal.
Mark the record appropriately, then click
Record and with the volunteer relaxed tap the
patellar ligament with the reflex hammer. Record 5 patellar reflex responses then click on
Stop. Measure the reflex time and magnitude
of each trial and place a representative screenshot of each measurement in your journal. Enter
the 5 reflex times, the 5 magnitudes and the
means in the worksheet. Measure the distance from
the black electrode to the L5-S1 vertebrae (top of hips)
and calculate the conduction velocity using the same
formula as you did for the Achilles reflex.
Record the data in the worksheet (Data Table 2).
C. Jendrassiks Maneuver
Figure 7. Electrode placement

for knee jerk reflex
B. Patellar Tendon Reflex
New electrodes should be placed with the
black (-1) lead placed about 12 cm from the
knee on the thigh overlying the quadriceps
muscle group. The red (+1) lead should be
placed about 10 cm above the black lead and
Revised Fall 2014
Repeat the patellar tendon reflex recordings on the

same volunteer while
the volunteer is pulling
their clenched hands
away from each other.
This is called Jendrassiks maneuver.
Repeat the calculations
of the mean reflex
time,
Figure 8. Jendrassiks maneuver
Page 43
magnitude and conduction velocities, enter the data

in the worksheet (Data Table 3) and copy representative recordings into the journal.
Revised Fall 2014
Page 44
Journal: __________

Date:
Name:
Section:
Achilles Tendon Reflex

1) Measure the reflex time (in msec) and magnitude of your 5 trials, calculate the means. then measure the path
length in mm and calculate the conduction velocity using the supplied formula.
Conduction Velocity ( m/sec) =
Total Path Length ( mm )

( M ean Reflex Time ( msec) - 0.5 msec )
Data Table 1 Achilles Tendon Reflex

Trial
Time (msec)
Magnitude (V)
1
2
3
4
5
Mean:
Path Length (mm):
Conduction Velocity
(m/sec)
Representative Vertebrate Conduction Velocities
Unmyelinated
1.2 m/sec
Myelinated
45 m/sec
2) Based on your data and the representative vertebrate conduction velocities shown in the above table, are the
nerves involved in your Achilles Tendon Reflex myelinated? Why is this adaptive for the organism?
_________________________________________________________________________________________
_________________________________________________________________________________________
_________________________________________________________________________________________
3) Describe the neural pathway involved in the Achilles tendon reflex (see the diagram on the previous page).
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
Revised Fall 2014
Page 45
Name:
(Continued)
4) Why is this reflex protective and what is it protecting?

___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
Patellar Tendon Reflex

5) Enter the reflex times, magnitudes, means, path lengths and conduction velocity in the table below :
Data Table 2 Patellar Tendon Reflex
Trial
Time (msec)
Magnitude (V)
1
2
3
4
5
Mean:
Path Length (mm):
Conduction Velocity
(m/sec)
6) Describe the neural pathway involved in the Patellar tendon reflex.
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
7) The reflex times for the Achilles Tendon Reflex and the Patellar Tendon Reflex should be different? Why?
___________________________________________________________________________________________
___________________________________________________________________________________________
8) Compare the conduction velocities of the two reflexes. What factors might explain your observations?
___________________________________________________________________________________________
___________________________________________________________________________________________
Revised Fall 2014
Page 46
Name:
(Continued)
9) Do you think the patellar reflex would be inhibited or enhanced by actively contracting the quadriceps muscle
group? Speculate on the mechanism of inhibition or enhancement?
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
Jendrassiks Maneuver
10) Enter the reflex times, magnitudes, means, path length and conduction velocities for the Patellar Tendon
Reflex when utilizing Jendrassiks maneuver:
Data Table 3 Patellar Tendon Reflex with
Trial
Time (msec)
Magnitude (V)
1
2
3
4
5
Mean:
Path Length (mm):
Conduction Velocity
(m/sec)
11) Enter your Mean reflex data in the summary table below:
Achilles Tendon Reflex
Patellar Reflex without

Patellar Reflex with Jendrassiks Maneuver
Mean Reflex Time (msec)

Conduction Velocity (m/sec)
Magnitude (V)
Revised Fall 2014
Page 47
Name:
(Continued)
12) Is the patellar reflex altered during Jendrassiks maneuver? If so, How and why do you think this might happen?
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
13) When describing the reflex arcs and the neural pathways involved we often limit ourselves to the nerves carrying information from the muscle or tendon to the spinal cord and back. However, it should be clear from our
ability to feel the tendon tap as well as the results from the Jendrassiks maneuver test that it isnt this simple.
What other neural connections must be present?
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
Revised Fall 2014
Page 48
Journal Format for Reflexes
(cont.)
Your Name
Lab Partners names
Laboratory Section
Date
Name: journal)
_______________
a complete
A. ACHILLES TENDON REFLEX

Paste the screen of a representative
myogram when bending the ankle
Paste a screen of a representative myogram showing the measurement of
reflex time when tapping the Achilles
tendon.
Paste a screen of a representative myogram showing measurement of the
magnitude when tapping the Achilles
tendon.
B. PATELLAR TENDON REFLEX
Paste the screen of a representative
myogram when bending the knee.
Paste a screen of a representative myogram showing the measurement of
reflex time when tapping the patellar
tendon.
Paste a screen of a representative myogram showing measurement of the
magnitude when tapping the patellar
tendon.
C. JENDRASSIKS MANEUVER
Paste a screen showing the measurement of reflex time while performing
Jendrassiks maneuver .
Paste a screen showing the measurement of reflex magnitude while performing Jendrassiks maneuver .
Revised Fall 2014
Page 49
Revised Fall 2014
Page 50
Biology 335 Human Physiology: Vertebrate Heart
Laboratory #8
Cardiology with a Vertebrate Heart
Objectives
1.
2.
3.
4.
To simulate the activity of the autonomic nervous

system by applying neurotransmitters directly to
an exposed heart.
To examine the effects of temperature on cardiac
contraction.
To examine the refractory period of myocardial
contraction.
To artificially induce heart block in an exposed
frog heart.
Background
receives input from both sympathetic (cardiac plexus) and parasympathetic (Vagus nerve) tracts. At
any given time, the heart rate and strength of contraction of heart muscle are influenced by the balance that
exists between these two sources of excitatory and
inhibitory innervation.
In this laboratory, after you record the frogs normal
heart action, you will alter the excitatory and inhibitory balance by applying various neurotransmitters and
receptor blockers directly onto the heart. Depending
on the substance that is applied, you will be simulating
activation of the sympathetic system or activation of
the parasympathetic system.
The heart of a frog looks

The neurotransmitters and recepvery much like that of a
tor blockers which will be used in
small mammals. A major
this laboratory can alter heart rate
internal difference exists,
by changing the rate at which the
however, in that the frogs
pacemaker of the heart (SA node
heart has only three chamin mammals or sinus venosus in
bers (Figure 1) while a
frog) generates action potentials.
mammals heart possesses
The normal rate of action potenfour. In mammals, the sitial generation is determined by
noatrial (SA) node, which is
the rate of spontaneous depolarilocated in the wall of the
zation which occurs between acright atrium, acts as the
tion potentials within the autonatural pacemaker. This is
rhythmic cells found within the
the structure that initiates each heart Figure 1. Amphibian Heart pacemakers. The alternating depolarizabeat. In the frogs heart, a portion of
tion and hyperpolarizations are called
the sinus venosus plays a similar role.
pacemaker potentials (Figure 2). The spontaneous
depolarization exhibited by autorhythmic cells is
Action potentials that originate within the pacemaker caused by a slow calcium current, the rate of which
(SA node or sinus venosus) travel via gap junctions
can be modulated by altering the potassium or sodium
through adjacent myocardial cells. The gap junctions permeability of the cells. Delaying the closure of poare part of the intercalated disks found at the junctions
between myocardial cells. Specialized myocardial
cells which provide preferential pathways for the
propagation of action potentials through these gap
junctions form the conduction pathways in the heart.
The cells of the conduction pathway are all capable of
generating pacemaker potentials and responding to
autonomic nervous system stimulation. However, in a
normal heart, the SA node (or sinus venosus in the
frog) is the pacemaker because the cells in this region
have the highest rate of action potential generation.
Cardiac function in vertebrates is regulated by the
autonomic portion of the nervous system. The heart
Revised Fall 2014
Figure 2. Pacemaker Potentials

Page 51

tassium channels
following an action
Student
potential will hyperpolarize the cell
When you receive the frog, you will note that
and lead to a slower
the animals spinal reflexes may still be inrate of depolarizatact. As part of the central nervous system,
tion between action
the spinal cord alone is capable of integrating
potentials and a
sensory (afferent) input and directing
slower heart rate.
appropriate muscular responses. The clasThis is how acetylsic example of such a reflex in humans is
choline and the parasnatching ones hand from a hot stove. Ansympathetic nervous
other is contraction of the quadriceps muscles
system cause a dewhen the patellar ligament is stretched by
Figure 3. Frog Heart Preparation tapping it with a rubber mallet. These involuncrease in heart rate.
Conversely, norepitary responses are innate and not learned; they
nephrine and the sympathetic nervous system induce
happen very
the opening of hyperpolarizing cyclic nucleotide
quickly without
(HCN) channels which increase the sodium current,
involvement of
causing the rate of depolarization to increase, thus
the brain. There
increasing heart rate
is awareness of
the situation
Action potentials (APs) originating within the SA
only after the
node (or sinus venosus) are propagated throughout the response has
heart via the conduction pathways mentioned previoccurred, as
ously. At the junction between the atria and the venmore time is
tricle there is a specialized collection of such cells
required for
called the atrioventricular (AV) node. Propagation of neural input to
APs through the AV node is delayed. This time delay reach the brain
is called the AV nodal delay and insures that atrial
and be integratcontraction proceeds prior to ventricular contraction.
ed by higher
From the AV node APs are propagated through the
brain centers.
Bundle of His, the bundle branches (left and right in
human) and finally through Purkinje fibers (cardiac
If the brain is
Figure 4. The Force Transducer
conduction cells) to each of the myocardial cells of the non-functional, as
heart.
it is in a these frogs, spinal reflexes may continue but
the animal cannot be aware of them. Innate reflexes
for a frog include retraction of the legs when they are
Preparation
touched. These muscular responses may be evident as
you secure the animal to the frog board.
Instructor
Place the frog dorsal side down on the board. Make
A few minutes before the beginning of the laboratory, an incision through the abdominal and thoracic walls
the instructor will anesthetize
the frogs by immersing them
in tricaine methanesulfonate. This anesthetic is
readily absorbed through the
skin of frogs. After the anesthetic takes effect, the frogs
brain will be destroyed.
From this time forward the
animals are brain dead and
cannot feel pain.
Figure 5. Acetylcholines effect on cardiac contraction.
Revised Fall 2014
Page 52

with scissors. You must cut through the skin and underlying musculature. You also need to cut through
the sternum to expose the thoracic cavity. Cut through
the ribs on either side of the sternum and remove the
sternum. The location of the heart should be readily
apparent, since it should be beating within its pericardial sac. Carefully cut through the parietal pericardium to expose the heart. BE CAREFUL NOT TO
CUT ANY MAJOR BLOOD VESSELS! If there is
already some blood in the body cavity, use a pipette or
paper towels to remove it so that you can see what you
are doing. Ringers solution may be used to rinse the
heart and surrounding structures if necessary. Pin the
fully extended forelegs firmly to the board (if necessary).
Identify the ventricle and the two atria (see Figure 1).
Take a bent pin or small fish hook and insert it
through the apex of the heart being careful NOT to
penetrate the lumen of the ventricle. Tie a 30 cm
length of thread to the end of this hook. Tie the other
end of the thread to the force transducer (see Figure
4). Adjust the tension on the thread so that it is taut.
The heart should be slightly elevated out of the thoracic cavity. Tension can be adjusted by moving the
transducer up or down on the ring stand.
Exercises
iWorx Setup
Calculating Heart Rate

We are using 5 beats to determine the heart rate.
If we divide the time for 5 beats by 5 this will
give us the time between each beat. If we then
divide 60 by that number we will be calculating
the number of beats in 60 seconds. These calculations can be simplified as shown below:
60
Time for 5 beats
5
5
60 X
Time for 5 beats
300
Time for 5 beats
=
=Beats/minute
mine the frogs heart rate the same way you determined
the pulse rate in the Introduction to iWorx laboratory
(see the box above) and measure the magnitude of a
normal contraction (measure the peak to trough height in
volts). Place a copy of each of these measurement
screens into your journal.
Note: If you have properly marked

your recording you can easily move
from exercise to exercise to compare the data by clicking on the
Marks icon and choosing the correct location from the list.
Start the LabScribe software

and choose 8 Cardiology
with a Vertebrate Heart and Vertebrate Heart from
the Settings menu. This will reveal a large window
labeled Frog Heart. Click Start in the LabScribe
software and make sure that you are getting a reasonably good record of the heart action on the computer.
You will probably have to click on the Autoscale button to get a decent recording. Stop recording when
everything is set up properly. Switch to the journal
and add your heading. Remember that there is a guide
to your journal format at the end of this chapter and
please remember to drag the left margin of your journal so that it fills one-half of the computer screen before you copy any screen images into it.
Effects of Cold and Warm on Cardiac

Function
A dropper bottle of saline solution cooled
in an ice bath will be used to apply cold
temperature to the heart.
Add an appropriate mark to the recording then begin

recording the heart contractions and apply 5-10 drops of
cold Saline solution to the heart. Measure the contraction rate and contractility (magnitude) and record each
of these screens in the journal. Did cold saline cause
any changes in heart rate or contractility?
Repeat the above procedure (after rinsing the heart with
room temperature saline) using Saline warmed in a water bath. Measure the contraction rate and contractility
(magnitude) and record each of these screens in the journal. Did warm saline cause any changes in heart rate or
contractility?
Effects of Neurotransmitters and Receptor Blockers
Normal Cardiac Contraction
Solutions containing epinephrine, acetylcholine and atRecord the action of the heart by clicking on Record.
ropine will be prepared by the instructor and shared by
Remember to add a mark to the data (maybe someall members of the class.
thing like normal contractions). Stop collecting data
after about 10 seconds. Click on Autoscale and deterRevised Fall 2014
Page 53

1.
EPINEPHRINE
Before applying epinephrine to the heart, record a few

heart beats to establish a baseline immediately prior to
drug treatment. Mark this pre-epinephrine. Then,
apply a few drops of the solution directly onto the
heart, aiming as best as you can for the sinus venosus
(the site of the pacemaker). This substance requires
some time to take effect, so wait about 2 minutes before recording the contractions. After recording the
contractions for about 10 seconds, compare this record
with the pre-epinephrine record to determine whether
heart rate or force of contraction (measured in volts
using the double cursors) has changed. If no change is
initially observed, apply additional epinephrine and
allow more time for it to take effect. When you
have a good response copy the pre-epinephrine and
post-epinephrine screens with the heart rate and
contractility measurements to your journal (4
screen shots).
By applying electrical stimuli to the heart muscle while

it is contracting you should be able to see when the heart
is capable of responding to stimuli and when it is refractory.
We will be using the iWorx/114 stimulator for this exercise. The electrode should be connected to the red portion of the iWorx front panel and held in place by a
clamp on your ring stand. Gently position the electrode
so that the two electrode wires make contact with some
region of the heart without impeding the hearts contraction. You should see a stimulator toolbar above the
Channel 1 screen on the computer. If the stimulator panel is not visible click on View and then Stimulator Panel. Towards the left end of this toolbar is a button labeled Apply. Make sure that the frequency setting is at
1. Whenever you click on the Apply button you will
stimulate the heart one time. This stimulus will appear
as a spike in the window labeled Stimulus.
2. ACETYLCHOLINE
Rinse the heart thoroughly with Saline solution to
eliminate any remaining epinephrine. Allow the heart
to normalize for about 10 minutes. Get a preacetylcholine record and then apply a few drops of
acetylcholine (ACh) to the right atrium. The effect is
usually rather rapid so you should start looking for a
response right away. Record the heart contraction for a
couple of minutes at least. Study the record for
changes in heart rate and/or strength of contracAmp=5 V
tion. If no change is observed after two
W(ms)=10
minutes, add additional acetylcholine and
F(Hz)=1
allow more time for it to take effect. When
#pulses=1
you have a good response, copy a tracing to
the journal and enter the heart rate and peak
heights which you measured.
3. ATROPINE
Rinse the heart again, but you do not need
to wait 10 minutes before applying a few
drops of atropine solution. Your ACh
record will serve as an indication of preatropine heart action. Record the hearts
response to atropine over the course of a
couple of minutes. Study the record for
changes in heart rate and/or strength of
contraction.
Refractory Period
Revised Fall 2014
Heart Block: Decreased conduction between the AV node and the

Bundle of His can disrupt coordination of atrial and ventricular contraction. First degree heart block is observed as an increase in the
time lag between atrial contraction and ventricular contraction.
Second degree heart block is seen as an occasional atrial contraction
which is not followed by a properly timed ventricular contraction.
Third degree heart block is a complete lack of coordination between
atrial and ventricular contraction.
Record a few normal contractions then stimulate the

heart using Apply at different times during the contractions. Your goal is to find a period of time during a
contraction when the stimulus has no affect on the
hearts contractions. Copy the screen showing this phenomenon into the journal.
Heart Block
Normal conduction of action potentials through the conduction pathways of the heart are important for the normal coordination of
cardiac contraction.
Especially important is
the slight time delay
built into the AV node
(AV nodal delay). In
this exercise we are
going to tie a ligature
Place ligature here
(thread) around the
heart between the atria
and ventricle (atrioventricular sulcus, see
Figure 6). By changing
how tight this ligature is
Figure 6. Position of Ligature
Page 54
we should be able to alter the conduction through the AV

node and mimic a pathological condition called heart
block.
Take a piece of thread about 30 cm long. Loop the thread
around the atria and tie a loose knot. Record a few normal
contractions then slowly tighten the knot (making sure that
the thread stays in the proper location).
Look at your recording. If there is no change you need to
tighten the ligature more until the atria and ventricles are
contracting independently of each other. This is called 3rd
degree heart block. Copy this recording into your journal.
Isolated Heart
Remove the heart from the frog by cutting through the
major blood vessels and any connective tissue remaining
around the base of the heart. Place the heart in a Petri dish
containing saline solution. Does it continue to beat now
that you have eliminated all nervous input to the heart? Is
it contracting at the same rate? Finally, separate the atria
from the ventricle by cutting through the AV septum with
a scalpel or razor blade. Are any of the individual pieces
of tissue still contracting?
Record your observations and answer the questions in the
Laboratory Worksheet.
Please remember to print or email a copy of the journal
for each lab member before you leave. Include the
journal format form and complete journal with your
worksheet.
Revised Fall 2014
Page 55
Revised Fall 2014
Page 56
Journal: __________

Date:
Name:
Section:
Cold and Warm

1) Record the heart rates and contraction strength (vertical displacement or V) during room temperature, cold
and warm stimulation in Data Table 1.
Data Table 1 Effects of Temperature
Treatment
Heart Rate (beats/

min)
Contraction Strength
(V)
Normal
Cold
Warm
2) On the graph below, plot the temperature (Normal, Cold and Warm) on the x axis and heart rate and contraction strength on the y axes. Use a bar graph and make sure that you label the graph appropriately. (Hint:
you can plot heart rate using the left axis and contraction strength using the right axis)
Revised Fall 2014
Page 57
Date:
Name:
Section:
3) Why does temperature alter heart rate and/or contractility?

___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
Neurotransmitters and Receptor Blockers

4) Record the heart rate and contraction strength before and after epinephrine, acetylcholine and atropine exposure in Data Table 2.
Data Table 2 Effects of Neurotransmitters
Condition or
Treatment
Heart Rate (beats/

min)
Contraction Strength
(V)
Pre-epinephrine
Epinephrine
Pre-Acetylcholine
Acetylcholine
Atropine
5) Explain the mechanism by which epinephrine increases heart rate.
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
6) Explain how epinephrine increases cardiac contractility.
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
Revised Fall 2014
Page 58
Date:
Name:
Section:
7) How does acetylcholine induce a decrease in heart rate?

___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
8) What is the mechanism by which acetylcholine causes a decrease in cardiac contractility?
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
9) What is atropine? What is its mechanism of action?
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
10) Describe the ionic cause of the prolonged cardiac contractile refractory period.
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
11) How is the refractory period of cardiac muscle different than that of skeletal muscle?
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
Revised Fall 2014
Page 59
Name:
Laboratory #7 Worksheet (cont.)
Name: _______________
(continued)
12) What role does the AV nodal delay play in normal cardiac function?
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
13) Did you notice any change in the ventricular heart rate when conduction between the atria and ventricles was
blocked by ligation? Explain this observation.
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
14) What does myogenic mean?
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
15) How did the different portions of cardiac tissue respond after being cut away from each other? Explain your
observations.
____________________________________________________________________________________________
____________________________________________________________________________________________
____________________________________________________________________________________________
____________________________________________________________________________________________
____________________________________________________________________________________________
Revised Fall 2014
Page 60
Name:
Laboratory #7 Worksheet (cont.)
Name: _______________
(continued)
16) Should epinephrine or acetylcholine alter the rate of contraction of the separated pieces? Explain.
____________________________________________________________________________________________
____________________________________________________________________________________________
____________________________________________________________________________________________
____________________________________________________________________________________________
____________________________________________________________________________________________
Revised Fall 2014
Page 61
Biology 335 Human Physiology: Frog Heart
Journal Format for Cardiology with a Vertebrate Heart
(cont.)
Your Name
Lab Partners names
Cardiology with a Vertebrate Heart
Laboratory Section
Date
Name: journal)
_______________
a complete
A. NORMAL CARDIAC CONTRACTION

Paste a screen of normal cardiac contractions with measurement of heart
rate.
Time for 5 beats = ______________ seconds
Heart rate (300/time for 5 beats) = _______________ beats/minute
Paste a screen showing the measurement of contraction strength.
Contraction Strength = _________ volts
B. EFFECTS OF COLD AND WARM ON CARDIAC CONTRACTION
Paste the screen showing the measurement of heart rate after cold saline.
Time for 5 beats = ________ seconds
Cold heart rate = __________ beats/min
Paste the screen showing the measurement of contraction strength after cold
saline.

Paste the screen showing the measurement of heart rate after warm saline.
Warm heart rate = __________ beats/min
Paste the screen showing the measurement of contraction strength after
warm saline.
Revised Fall 2014
Page 62
Journal Format for Cardiology with a Vertebrate Heart (continued)
(cont.)
(guide for
setting
_______________
up Name:
your journal)
C. EFFECTS OF NEUROTRANSMITTERS AND RECEPTOR BLOCKERS

Epinephrine
Paste a screen of the measurement of
heart rate pre-epinephrine.
Paste a screen showing the measurement contraction strength preepinephrine
heart rate post-epinephrine.
Paste a screen showing the measurement contraction strength postepinephrine
Acetylcholine
Paste the screen showing the measurement of heart rate pre-acetylcholine.
Heart rate = __________ beats/min
Paste the screen showing the measurement of contraction strength preacetylcholine.

Revised Fall 2014
Page 63
Journal Format for Cardiology with a Vertebrate Heart (continued)
(cont.)
(guide for
setting
_______________
up Name:
your journal)

heart rate post-acetylcholine.
Paste a screen showing the measurement contraction strength postacetylcholine
Atropine
Paste the screen showing the measurement of heart rate post-atropine.

Paste the screen showing the measurement of contraction strength postatropine.
D. REFRACTORY PERIOD
Paste a screen clearly indicating at
least one stimulus applied during the
refractory period.
E. HEART BLOCK
Paste a screen showing disruption of
the cardiac cycle due to ligation and
damage to the AV node.
Revised Fall 2014
Pa
Biology 335 Human Physiology: EKG
Laboratory #9
Electrical Properties of the Heart
pulmonary veins (and thus within the atria) and the
atrioventricular valves will be forced closed. The closure of these valves causes turbulence and vibrations
1. To understand the origins of normal and abnormal within the blood and the great vessels. These vibraheart sounds.
tions can be heard at the chest wall as the first heart
2. To take blood pressures and pulse rates.
sound (lubb or S1). As the contraction ends, the
3. To record electrocardiograms.
heart will enter a resting period called diastole. As
4. To examine the effects of exercise on these meas- the heart relaxes, the pressure within the ventricles
urements.
will drop below the arterial pressure in the pulmonary
arteries and aorta. When this occurs the semilunar
Cardiac Auscultation
valves will close producing the vibrations which we
hear as the second heart sound (dubb or S2). We can
hear these sounds using a stethoscope. [The following
Background
link: http://depts.washington.edu/physdx/heart/
demo.html will bring you to a website which provides
In mammals (including humans), the normal cardiac
examples of both normal and abnormal (murmurs)
cycle occurs with the rhythmic opening and closure of heart sounds.]
the four heart valves as a consequence of the hearts
rhythmic contraction. These valves include the right
Procedure
atrioventricular valve (tricuspid), the left atrioventricular valve (bicuspid), the pulmonary semilunar and the We will use the iWorx 214 equipaortic semilunar
ment and an electronic stethovalves. The atrioscope to record heart sounds as
ventricular valves
we listen to them at the base and
are located at the
apex of the heart.
entrances to the
ventricles while
Insert the mini DIN plug from
the semilunar
CH4 of the iWorx 214 unit into
valves are located
the jack on the side of the ES100
at the exits. The
stethoscope (Figure 3). Turn on
opening and clothe iWorx 214 and start the Labsure of these
scribe 2 software. Under Setvalves occurs
tings, Load Group click on 9
because of hydroElectrical Properties of the
Figure 2. Stethoscope placement
static pressure
Heart then under the Settings
differentials
menu click on Electrical Properties of the Heart.
which occur within Figure 1. Stethoscope and BP
You should now see one window labeled Heart
Cuff
the heart and the
Sounds.
great vessels (vena
According to the American Heart Association, Korotkoff's
cavae, aorta, pulmonary arteries, pulmonary veins).
Objectives
During cardiac contraction (systole) the blood pressure within the

Korotkoff,
Nikolai
Sergieleft and right venyevich (b 1874), Russian physitricles will exceed
cian. Korotkoff introduced the
the blood presauscultation method of detersure within the
mining blood pressure in 1905.
vena cavae and
Revised Fall 2014
sounds occur in five phases.

Phase 1: faint, clear, tapping sounds. This is the systolic
pressure.
Phase 2: murmurs or swishing sounds
Phase 3: crisper, more intense sounds
Phase 4: distinct, abrupt muffle of sound. In children, this is
the diastolic pressure; in adults, it reveals hyperkinetic state
(increased movement in blood vessels from disease or strenuous exercise) if it remains throughout deflation.
Phase 5: no longer any sound. This is diastolic pressure in
adults
Page 65
Blood Pressure
Background
Figure 3. iWorx 214 with ECG leads

and ES100 electronic stethoscope.
Figure 4. Recording of heart sounds with ES100.

1.
2.
3.
Use the ES100 electronic stethoscope

(instructions for the use of the stethoscope are
provided) to listen to a partners heart sounds at
the apex (bottom) and at the base (top) of the
heart (see Figure 2) while recording the sounds
with the iWorx. Please note that you must hold
the stethoscope very steady to avoid noise.
Copy screen shots of the recordings at the base
and apex into your journal (see Figure 4). Label
S1 and S2.
As you listen to the heart sounds attempt to discriminate between the first and second heart
sounds and attempt to determine the difference
between systole and diastole.
Answer the questions on the worksheet.
Revised Fall 2014
As the heart contracts it produces pressure waves

which travel through your blood vessels. During cardiac contraction (systole) this hydrostatic pressure
reaches a high point and during cardiac relaxation
(diastole) it reaches a low point. These pressures are
referred to as the systolic and diastolic blood pressures. They are easily measured using a blood pressure cuff and sphygmomanometer. The blood pressure cuff is used to occlude the blood vessels in a persons arm while the sphygmomanometer (sphygmo
= artery) measures the pressure exerted by the cuff.
Once the blood flow is completely occluded by the
blood pressure cuff, the pressure exerted by the cuff is
gradually decreased. When the pressure exerted by
the cuff is less than the systolic blood pressure, blood
will be forced past the cuff in a pulsatile fashion setting up vibrations which can be heard using a stethoscope placed just downstream from the occluding
cuff. This pressure represents the pressure of blood
flowing through the arteries of the arm while the heart
contracts. As the pressure is allowed to decrease even
more, you will eventually hear a whoosh and then
silence as blood flow is no longer hindered. This occurs as blood is allowed to flow freely through the
blood vessels. These sounds are called Korotkoff
sounds. In this part of the laboratory we will record
pulse waves in the finger and correlate them with the
korotkoff sounds as recorded using the ES100 stethoscope. In the second part of this exercise you will
determine the effects of exercise upon blood pressure
and cardiac output of a volunteer.
Recording Pulse and Korotkoff Sounds
Procedure
In the Labscribe software click on the Edit menu then
click on Preferences. This will bring up a screen
which determines what you will be recording and the
inputs that are being utilized. Make sure that both Ch
2 Pulse and Ch 4 Heart Sounds are checked and that
EKG is NOT checked, then click on OK. You should
now see two windows. The top window is labeled
Pulse and the lower window is labeled Heart
Sounds.
1. Attach a blood pressure cuff to the arm of a volunteer in your lab group, place the pulse oximeter
on the thumb of the same hand, and place the bell
of the ES100 electronic stethoscope in the antePage 66
Pulse pressure (mm Hg) = systolic BP diastolic BP

Stroke volume (mL) = pulse pressure x
1.7
Cardiac Output (mL/min) = stroke
volume x pulse rate
Record these data in your worksheet.
Stroke volume and Cardiac output are
very important parameters of cardiovascular function. Stroke volume refers to
the amount of blood pumped by each
ventricle with each contraction. Cardiac
output represents the total amount of
blood pumped by each ventricle in a
minute.
Figure 5. Pulse and Korotkoff sounds recording during blood
pressure measurements.
cubital region of that same arm.
Make sure the stethoscope is turned on and
plugged into the iWorx 214 unit.
3. Click on Record then increase the pressure in the
cuff until blood flow in the finger is halted (you
can see this in the Pulse window (see figure 5).
Slowly release the pressure from the blood pressure
cuff while listening for the Korotkoff sounds. Your
recording should look like Figure 5. Copy a screen
shot of your recording in the journal. Label the events
that you recorded.
2.
Heart Rate and Blood Pressure Before and After

Exercise.
Procedure
1.
2.
3.
4.
5.
Establish a baseline resting blood pressure and

heart rate (use the pulse rate taken from the subjects radial artery) for your volunteer. Record
these data in the worksheet.
The volunteer should exercise vigorously for 2
minutes (jumping jacks work fine).
Immediately following exercise (0 minutes), the
volunteers blood pressure and pulse should be
recorded again.
Repeat the measurements one, three, and five
minutes later. (Alternate arms between measurements)
For each measurement, calculate the pulse pressure, stroke volume and cardiac output.
Electrocardiogram
Background
An electrocardiograph is an
instrument that allows an
investigator or clinician to
obtain a record of electrical events that occur during the cardiac cycle.
Several electrodes, placed
at different locations on the Figure 6. Standard Limb Leads
surface of the body, are
used to detect electrical activity that originates within
the heart. The recording obtained, an electrocardiogram (ECG or EKG), represents a plot of the voltage
difference measured between any two of these electrodes (Y axis) against time (X axis). The specific pair
of electrodes being used to produce a recording is referred to as a Lead. Lead I records the voltage between the electrodes located on the left arm (LA) and
right arm (RA); Lead II records the voltage between the
left leg (LL) and right arm (RA); Lead III records the
voltage between the left leg (LL) and left arm (LA).
These three standard leads (I, II and III) use only two
electrodes at a time. Polarity between the two contact
points is specified in such a way that the investigator
knows which way the pen on the recorder will move
relative to a zero potential. For example, with Lead I,
when the LA is positive relative to the RA, the pen will
Page 67
Revised Fall 2014

This laboratory procedure was adapted from DeCoursey, R.M. and F. Dolyak, Laboratory Manual of
Human Anatomy and Physiology, 3rd ed., New York,
McGraw-Hill, 1974.

deflect above zero. Lead II is
defined as LL positive relative to
RA, and Lead III is defined as LL
positive relative to LA. Standardization among all electrocardiographs is the objective in defining polarity. In addition to these
standard leads, augmented leads
can be used to produce electrocardiograms. These leads are
designated AVR, AVL and
AVF. With these leads, three
electrodes (LA, RA and LL) are
used, and the last letter in each
designation defines polarity, For
AVR, the right arm (RA) is positive relative to the other two electrodes. For AVL the left arm
(LA) is positive; for AVF the left
leg is positive. These can easily
be remembered if you keep in
mind that R and L stand for
right and left sides of the body,
while "F" can be related to a
foot which is associated with
the left leg.
T2-T1
1
Figure 7. Measuring Heart Rate
Although an electrode is customarily attached to the

right leg, it is not part of any of the standard or augmented leads described above. Chest leads are commonly used in clinical settings and provide a great
deal more information about the electrical activity of
the heart. In todays laboratory we will just be using
Limb Lead I with the iWorx 214 apparatus.
Procedure
We will use our iWorx 214 equipment to obtain electrocardiograms before and after exercise in order to
examine the electrical activity of the heart under different conditions.
Make sure the EKG leads are connected to the iWorx
214 apparatus as shown in Figure 3. Click on Edit
then on Preferences and in then check the EKG and
Pulse channels. Make sure that you uncheck the
Heart Sounds channel. Click on the mode/
function of the EKG channel and set it to 0.03-150
Hz. Adjust the windows so that you have a large
EKG recording area and a smaller screen beneath
labeled Pulse as shown in Figure 7.
This need not be the same subject as used for the blood
pressure exercise, but it must be a person able to do the
2 minutes of exercise. The subject should remove his/

her watch and any other jewelry that might contact and
interfere with the electrode.
Attach the electrodes to both wrists and the left ankle of
your volunteer lab partner as follows:
Red = right wrist
Black = left wrist
Green = left ankle
During recording, the subject must be quietly seated
and sit away from the lab bench without moving. This
will minimize electrical interference. When the subject
is sitting quietly, click the Record button (after placing
a mark onto the record) and record for approximately
15 seconds. Copy this recording into the journal
(please remember to enter the heading). From these
data you can easily measure heart rate, P-R interval and
ventricular systolic and diastolic times.
Heart rate: Measure the time it takes for 5

cardiac cycles (see Figure 7). Divide 300
by this number to give you the heart rate
in beats/minute.
P-R interval: Using the double cursors,
measure the time from the onset of the P
wave to the onset of the next Q (or R)
Page 68
Revised Fall 2014

McGraw-Hill, 1974.
wave (see figure 8).

V-systole: Measure the time from the
peak of one R wave to the peak of the
next T wave (see figure 9).
V-diastole: Measure the time from the
peak of the T wave to the peak of the next
R wave (see figure 10).
Now ask your subject to do jumping jacks or other

exercise vigorously for two minutes. You should
leave the electrodes in place on the wrists and ankle,
but DETACH THE WIRES FROM THE ELECTRODES. This allows the subject freedom of movement, prevents damage to the apparatus, and allows
you to hook it all back up quickly when the exercise is
completed. As soon as possible, obtain another record . Repeat the measurements you performed on the
pre-exercise data and copy all of this into the journal.
Figure 8. P-R interval
Pulse Delay
Place the pulse oximeter on a finger of the volunteer
then with your volunteer sitting quietly record the EKG
and pulse simultaneously for about 15 seconds. Copy
this record into your journal and measure the time delay between peak of the QRS complex and the onset of
the pressure pulse in the finger (see figure 11).
Figure 9. V-systolic time
See the Journal Format Form Provided at

the End of the Chapter!
Figure 10. Diastolic time
Figure 11. Pulse delay

Page 69
Revised Fall 2014

McGraw-Hill, 1974.
Revised Fall 2014
Page 70
Journal: __________

Date:
Name:
Section:
Cardiac Auscultation
1) Describe the differences you heard between the heart sounds when you listen at the base compared to the
apex of the heart.
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
2) Could you detect any abnormalities in your volunteers heart sounds? If so, describe the sounds.
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
Blood Pressure
Remember:
Pulse pressure = systolic-diastolic BP
Stroke volume = pulse pressure X 1.7
Cardiac Output = heart rate X Stroke volume
3) Calculate the cardiac outputs and place the data in the following table:
Time
Heart Rate
(b/min)
Systolic BP
(mm Hg)
Diastolic BP Pulse Pressure Stroke Volume

(mm Hg)
(mm Hg)
(ml)
Cardiac
Output
(ml/min)
Resting
0 minutes
post-exercise
1 minute post
-exercise
3 minutes
post-exercise
5 minutes
post-exercise
4) Using the graph on the next page, plot the cardiac output on the y axis and time on the x axis.
Revised Fall 2014
Page 71
Revised Fall 2014
Page 72
Date:
Name:
Section:
5) What do your data tell you about the effects of exercise on heart rate, systolic BP, diastolic BP, and cardiac
output?
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
6) What happened to the cardiac output just after exercise and during recovery from exercise?
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
Electrocardiogram
7) Label the P, QRS and T waves on a normal cardiac cycle in your journal.
8) Enter the pre and post-exercise heart rate and
EKG information in the table. Calculate the difference between the pre- and post-exercise data
and the percent difference. Please make sure that
your measurement screens are included in the
journal as well.
9) How does the subjects pre-exercise P-R interval
compare to a normal interval of 120-200 msec?
Parameter
PreExercise
PostDifference % Difference
Exercise (Pre-Post (Difference/
Exercise Pre-Exercise)
*100
Heart rate
(beats/min)
P-R Interval
(msec)
__________________________________________ V-systole
(msec)
__________________________________________
V-diastole
__________________________________________ (msec)
___________________________________________________________________________________________
Revised Fall 2014
Page 73
Date:
Name:
Section:
10) What does a P-R interval greater than 200 msec mean for a patient?
___________________________________________________________________________________________
___________________________________________________________________________________________
11) Why would you expect diastole to be longer than systole for someone with a resting heart rate?
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
12) Assuming that your subjects heart rate increased, this means that each cardiac cycle must be completed in a
shorter period of time. This could be accomplished by shortening systole, diastole, or both. Which phase of
the cardiac cycle shortened the most?
________________________________
13) Why do you think that shortening of this part of the cycle does not seriously hinder ventricular filling?
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
14) What was the time delay between the QRS complex and the onset of the pressure pulse in your subjects
finger?
________ msec
15) Do you think this delay would change if you measured the pulse using a toe instead of a finger? Explain.
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
Revised Fall 2014
Page 74
Biology 335 Human Physiology: ECG
Journal Format for Electrical Properties of the Heart
(cont.)
Your Name
Lab Partners names
Electrical Properties of the Heart
Laboratory Section
Date
Name: journal)
_______________
a complete
CARDIAC AUSCULTATION
Paste a screen showing a recording of

heart sounds from the base of the heart.
Label S1 and S2.

heart sounds from the apex of the
heart. Label S1 and S2.
BLOOD PRESSURE

the pulse waves and Korotkoff sounds
when measuring blood pressure. Label
the events on the recording.
Revised Fall 2014
Page 75
(cont.)
Name: journal)
_______________
a complete
ELECTROCARDIOGRAM
Pre-Exercise EKG
Paste a screen showing the measurement and calculation of heart rate.

Clearly mark the P, Q, R, S and T
waves on one of the cycles.
T
P
Q S

Heart rate (300/time for 5 beats) = _______________ beats per minute
Paste the screen showing the measurement of pre-exercise P-R interval.
Pre-exercise P-R interval = __________ seconds

Paste the screen showing the measurement of pre-exercise ventricular systolic time.
Pre-exercise ventricular systolic time = _________ seconds
Paste the screen showing the measurement of pre-exercise ventricular diastolic time.
Pre-exercise ventricular diastolic time = __________ seconds
Revised Fall 2014
Page 76
(cont.)
(guide for
setting
_______________
up Name:
your journal)
Post-exercise EKG
Paste a screen showing the measurement and calculation of post-exercise
heart rate.

Heart rate (300/time for 5 beats) = _______________ beats per minute
Paste the screen showing the measurement of post-exercise P-R interval
Post-exercise P-R interval = __________ seconds
Paste the screen showing the measurement of post-exercise ventricular systolic time.
Post-exercise ventricular systolic time = _________ seconds

Paste the screen showing the measurement of post-exercise ventricular diastolic time.
Post-exercise ventricular diastolic time = __________ seconds

Pulse Delay
Paste the screen showing the measurement of pulse delay.
Pulse delay = _____________ seconds
Revised Fall 2014
Page 77
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Page 78
Biology 335 Human Physiology: Circulation
Laboratory #10
Circulatory Physiology
pulsatile. As blood enters capillaries, it will flow
more slowly and in a non-pulsatile manner. The diameter of a capillary is just slightly greater than that
1. Observe living capillary beds and understand how of a red blood cell, so blood cells pass through capilblood flow through a capillary bed is regulated.
laries in single file. The walls of capillaries are very
2. Explore the physiological control of blood flow to thin. Venules, which receive blood that leaves capilhuman skin.
lary beds, are the most difficult to identify with certainty, but these are larger in diameter than capillaries
and blood flow within them is non-pulsatile. Blood
Microcirculation
normally flows from arterioles to capillaries to venules (two exceptions include the hepatic portal and
hypothalamo-hypophyseal
Background
portal systems).
Objectives
The purpose of the first part

Capillary function depends
of this laboratory is to obupon blood flow. If more
serve microcirculation.
blood is flowing through a
Microcirculation refers to
capillary bed, more oxygen
blood flow in the smallest
will be delivered to that tisblood vessels within the
sue. Within capillary beds,
body -- arterioles, capillarthere is at least one through
ies, and venules. It is usualfare channel (Figure 1)
ly very difficult to observe
which provides a preferenblood flowing in these vestial avenue for blood flow
sels, because most of these
through the tissue. There
networks, centered around
are also a number of other
capillary beds, are located
capillaries penetrating into
deep within organs. Howthe tissue whose function is
ever, there are a few anadependent upon the quantity
tomical situations that are
of blood flowing into the
particularly conducive to
capillary bed. VasoconFigure 1. Capillary Bed
such study -- specifically wing memstriction or vasodilation of the arterioles
branes of bats and the webs between the toes of frogs. or the precapillary sphincters can dramatically alter
Both of these structures consist of a sandwich of
the quantity of blood flowing through a capillary bed.
two epithelia
These alterations in blood flow will alter the number
with a layer of vascular connective tissue between.
of capillaries which are actually carrying blood at a
Because each of these sandwiches is so thin, blood
given time. Changes in blood flow into a capillary
flow in the connective tissue can be observed simply
bed are thus most easily detected as changes in the
by shining light through the wing or web and observ- number of functioning capillaries.
ing with a microscope.
Vasoconstriction is physiologically regulated by neuToday you will work with frogs. Following the inral, endocrine and local metabolic mechanisms. The
structions below, obtain the best possible view of
neural regulation of vasoconstriction involves the
blood flowing through the small blood vessels in the
sympathetic nervous system stimulation of the vascuweb of the foot. You should be able to distinguish
lar smooth muscle of arterioles by norepinephrine
arterioles, capillaries, and venules. Arterioles have
released by autonomic motor neurons. The endocrine
muscular walls and are larger in diameter than capilregulation involves the release of epinephrine from
laries; blood flow through arterioles often appears
the adrenal medulla in response to activation of the
Revised Fall 2014
Page 79
Magnification dial
Focusing dial
Figure 3. Wrapped frog on stage
Power
Lighting
controls
Stage
Figure 2. Stereomicroscope
sympathetic nervous system originating in the medulla

oblongata of the brain. Local controls include temperature and the levels carbon dioxide, oxygen, pH, histamine and other chemicals within the tissues. In this
laboratory, local effects will be examined by bathing the
frog foot in warm or cold saline solution. Autonomic
nervous system and endocrine effects will be examined
by applying epinephrine (parasympathetic inputs do
not normally alter vasoconstriction) to the tissue while
the effects of local modulators will be examined by
applying histamine to the frog foot.
The arterioles in the skin express alpha-1 adrenergic receptors while arterioles in skeletal muscle
express beta-2 receptors. The alpha-1 receptors are
excitatory while beta-2 receptors are inhibitory.
When norepinephrine (or epinephrine) bind to these
receptors the simultaneously cause vasoconstriction
in the peripheral arterioles and vasodilation of skeletal muscle arterioles. This means that activation of
the sympathetic nervous system initially will cause
peripheral vasoconstriction and skeletal muscle
vasodilation increasing blood flow to skeletal muscle and decreasing blood flow to the skin (and viscera).
Revised Fall 2014
Exercise causes an immediate increase in sympathetic

nervous system activity which is a consequence of
sympathetic stimulatory inputs from the primary motor cortex of the brain to the medulla oblongata autonomic nervous system control centers. The medulla
oblongata also receives input from hypothalamic thermoregulatory centers. Increases in core body temperature are detected by thermoreceptive cells in the hypothalamus and can alter the autonomic control centers in the medulla and cause decreased sympathetic
output to the peripheral vasculature thus causing vasodilation and increased blood flow to the skin. This
increased flow is instrumental in increasing radiated
heat loss (in conjunction with sweating) and thus
cooling of the core body temperature.
Procedure
The instructor will immobilize the frogs by placing
them in a container containing tricaine methane sulfonate. This substance is an anesthetic that is ab-
Figure 4. Frog Foot

Page 80
sorbed across the skin. When the frog no longer responds to touch (this requires at least 15 minutes) the
instructor will remove it from the anesthetic and wrap
it in a paper towel that has been soaked in tap water,
leaving its hind feet exposed. This will prevent the
animals skin from drying out while you are observing
the circulation.
ed in the worksheet. Remember that generally, epinephrine is considered a vasoconstrictor while histamine is a vasodilator.
Exercise and the Skin

Background
The circulation within the webbing of the foot will be

observed using a Stereomicroscope (Zoom or Dissecting microscope) as shown in Figure 2. The frog
wrapped in a moistened paper towel should be placed
ventral surface down on the stage of the microscope
with one foot over the window in the base (Figure
3). Spread the toes in such a way that the toes are
widely separated, with the webs between the toes being as flat as possible (Figure 4).
The purpose of this experiment is to determine how

exercise affects blood flow to the skin in human subjects. Chose as your subject a member of your group
who can safely do at least 5 continuous minutes of
moderate exercise.
Using the highest magnification of the dissecting microscope (350X), identify arterioles, capillaries, and
venules. Observe blood flow through these vessels
and get a feel for how differently it flows through
each type of vessel. Remember that blood flows
from arterioles into capillaries and then into venules.
Temperature Effects
Drip some warm saline onto the web. Is blood flow
faster or slower? Do the arterioles in your field of
view dilate or constrict? Do you think the blood flow
(volume) is higher or lower? Describe what you see
in the space provided in the worksheet.
The skin is the largest organ of the human body and
contains the largest reservoir of blood in the body. It
is also extremely important in regulating body temperature (thermoregulation). As a blood reservoir, increased metabolism within other tissues will cause a
decrease in blood flow to the skin as it is diverted to
supply those tissues with oxygen, etc. As a therEpinephrine and Histamine
moregulatory organ, increased core body temperature
Allow the circulation to return to normal by allowing will cause an increase in blood flow to the skin to inthe foot to warm up to room temperature (but dont let crease radiated and evaporative heat loss. These two
functions (blood reservoir and thermoregulation) can
it dry out). Then, test the effects of epinephrine and
histamine on the microcirculation. First, drip some of be easily observed during exercise.
the epinephrine solution (available on the front desk)
onto the web and record in the space below any chang- In this exercise we will be measuring the blood flow
es you observe. Be sure to allow enough time for the through a finger/thumb using a pulse oximeter. These
devices detect pressure changes in the finger and using
solution to soak into the tissue. After each treatment record your observations in the space provid- the LabScribe software we can automatically display
Apply some ice water to the web. What is happening
now? Describe what you see in the space provided
in the worksheet.
Revised Fall 2014
Page 81
the integral of the pressure change which is a relative

measurement of the blood flow through the finger
(peripheral blood flow).
Procedure
Place the pulse oximeter on the index finger or thumb
of your volunteer. Use the same digit each time a
measurement is obtained. In the Settings menu Load
10 Circulation, then under settings choose 10 Circulation. One channel is titled Pulse, the second is labeled % O2 and the other is titled Blood flow. Record
the normal resting blood flow of your volunteer (be
sure to mark the recording appropriately) and save this
to your journal (dont forget to start with your heading). Repeat the measurement for 5 separate pulses,
record the data in the data table in the worksheet. Calculate the mean blood flow.
Send the subject off to exercise for 1 minute. When he
or she returns, record the pulse and blood flow as
above (5 measurements) and record the data in the data
table and copy a screen shot in your journal. Finally,
as soon as possible, have the subject do 3 more
minutes of moderate vigorous exercise and repeat the
measurements. If there are no differences you may
have to repeat the measurements after an additional 2
minutes of exercise.
Blood flow can then be determined by measuring the
trough to peak difference [see below] (V2-V1=ml/
min) for each of the time points (0, 1, 3 and possibly 5
minutes of exercise).
As always these data need to be copied to the journal
and saved. Before you leave you should perform all of
the measurements and print a copy of your journal for
Revised Fall 2014
Page 82
Journal: __________


Date:
Name:
Section:
Microcirculation
1) Describe how blood flows through the microcirculation of a typical tissue. Include brief descriptions of the
different blood vessel types which are involved.
_________________________________________________________________________________________
_________________________________________________________________________________________
_________________________________________________________________________________________
_________________________________________________________________________________________
_________________________________________________________________________________________
2) What observations can you make concerning blood flow through the different types of vessels in the frog foot?
_________________________________________________________________________________________
_________________________________________________________________________________________
_________________________________________________________________________________________
3) What did you observe when you applied warm water to a capillary bed?
_________________________________________________________________________________________
_________________________________________________________________________________________
4) What happened when you applied cold water?
___________________________________________________________________________________________
___________________________________________________________________________________________
5) How can you detect vasoconstriction or vasodilation in the frogs foot using low magnification stereo microscopes?
_________________________________________________________________________________________
_________________________________________________________________________________________
6) Which treatments caused vasoconstriction of blood vessels in the frogs skin?
____________________________________
_______________________________________
7) Which treatments caused vasodilation of blood vessels in the frogs skin?

____________________________________
Revised Fall 2014
_______________________________________
Page 83

Laboratory #9 Worksheet (cont)
Name:
Name: _______________
(continued)
8) According to Poiseuilles Law, vasodilation is associated with an increase in blood flow through a capillary bed.
How do your observations support this concept?
________________________________________________________________________________________
________________________________________________________________________________________
________________________________________________________________________________________
Blood Flow and Exercise

9) Record your blood flow data in the table provided.
Blood Flow (ml/min)
10) On the following page, graph the mean blood flow on

the y axis and exercise time on the x axis.
11) Describe the effects that exercise had on the peripheral
circulation of your subject at each time point.
1
_____________________________________________
_____________________________________________
_____________________________________________
Before
Exercise
1 min of
exercise
3 min of 5 min of
exercise exercise
2
3
4
5
12) What is the physiological significance of vasoconMean

striction and/or vasodilation of the peripheral circulation during exercise? When during exercise might they occur?
________________________________________________________________________________________
________________________________________________________________________________________
_________________________________________________________________________________________
13) How does the brain control peripheral vasoconstriction and vasodilation?
________________________________________________________________________________________
________________________________________________________________________________________
________________________________________________________________________________________
14) During exercise the same neurotransmitter can cause vasodilation in one tissue and vasoconstriction in a different tissue. How does this work?
________________________________________________________________________________________
________________________________________________________________________________________
_________________________________________________________________________________________
Revised Fall 2014
Page 84
Revised Fall 2014
Page 85

Name:
Name: _______________
(continued)
15) Most immediate physiological adjustments to exercise occur prior to any change in tissue metabolic demand
for oxygen or nutrients. This occurs because exercise increases sympathetic nervous system activity. With your
knowledge of exercise and the control of alpha motor neurons by the primary motor cortex of the brain, how does
increased exercise influence the sympathetic nervous system?
________________________________________________________________________________________
________________________________________________________________________________________
________________________________________________________________________________________
________________________________________________________________________________________
________________________________________________________________________________________
________________________________________________________________________________________
________________________________________________________________________________________
Revised Fall 2014
Page 86
Journal Format for Circulation
Laboratory #4
Worksheet
(cont.)
(Your
journal should
contain
Your Name
Lab Partners names
Circulation
Laboratory Section
Date
theName:
following_______________
components)
Paste a screen showing the measurement of pre-exercise blood flow.
Mean blood flow before exercise = _______________________

Paste a screen showing the measurement of blood flow after 1 minute of
exercise.
Mean blood flow after 1 minute of exercise = _______________________
Paste a screen showing the measurement of blood flow after 3 minutes of
exercise.
Mean blood flow after 3 minutes of exercise = _______________________
Paste a screen showing the measurement of blood flow after 5 minutes of
exercise.
Mean blood flow after 5 minutes of exercise = _______________________
Make sure all 4 journal pages are in order and turn them in with the worksheet
Revised Fall 2014
Page 87
Revised Fall 2014
Page 88
Biology 335 Human Physiology: Breathing
Laboratory #11
Mechanisms of Breathing
Objectives
text to re-familiarize yourself with the muscles involved in breathing.
1.
2.
3.
The changes in thoracic volume (which will alter lung

volume) that occur during the ventilation can be estimated by measuring the changes in thoracic and abdominal diameters and widths during inhalation and
exhalation. These changes in thoracic diameter should
reflect actual changes in thoracic cavity volume which
will cause changes in intra-alveolar pressure and
changes in lung volume.
4.
Examine the mechanics of ventilation.

Measure lung volumes of a human volunteer.
Examine the effects of exercise on lung volumes
and respiratory rhythmicity.
Explore the mechanisms underlying the regulation
of respiration.
Respiration
Changes in lung volume can be measured using a spirometer. A spirometer is a device which measures the
Background
volume of air being inhaled or exhaled while a subject
breathes. The most important of these volumes are:
Respiration is commonly considered under two head- Tidal volume (TV, see volume a in figure 1)the
ings internal respiration and external respiration.
volume of a normal resting breath (normally
around 500 mL).
Internal or cellular respiration is concerned with the Inspiratory Reserve Volume (IRV, see volume c
physical and chemical factors involved in the utilizain figure 1) the volume which can be inhaled
tion of oxygen and formation of carbon dioxide by
in addition to the normal TV.
tissue cells. Mitochondria contain the enzymes that
Expiratory Reserve Volume (ERV, see volume b
catalyze the chemical reactions of cellular respiration.
in figure 1) the volume which can be exhaled
in addition to the normal TV.
External respiration includes:
Residual Volume (RV, see volume e in figure 1)
the volume of air left in the lungs after a maxi1) Breathing (or ventilation of the lungs): the gasemal exhalation.
ous exchange between an organism and its envi Vital Capacity (VC, see volume d in figure 1)
ronment, which provides for maintenance of an
the maximum volume of air which can be exadequate oxygen supply in alveolar air and elimination of carbon dioxide. Breathing requires the
action of the diaphragm and other thoracic and
abdominal muscles. The coordination of these
muscles is regulated and controlled by the nervous system.
2) Exchange of oxygen and carbon dioxide between
alveolar air and the blood within lung capillaries.
3) Transport of oxygen and carbon dioxide by the
blood between the lungs and metabolizing tissues.
Breathing is characterized by the bulk flow of air into
and out of the lungs. This flow is driven by air pressure changes within the thoracic cavity which occur
because of changes in the volume of the thoracic cavity. Such volume changes are accomplished via the
abdominal and thoracic musculature, primarily the
diaphragm. You should quickly review an anatomy
Revised Fall 2014
Figure 1. Lung Volumes

Page 89
changed in a single breath.

Total Lung Capacity (TLC, d+e) = VC + RV
The rate at which air can be expelled from the lungs is

a useful clinical tool in diagnosis of obstructive pulmonary diseases. Normally, a person should be able
to expel between 75% - 85% of his/her vital capacity
in the first second of a forced exhalation. This measurement is called the FEV1. If airways are obstructed,
however, as in asthma, exhalation is hindered and the
FEV1 will be lower.
ing blood carbon dioxide level can alter the breathing

rhythm.
Thoracic and Abdominal Dimensions

Procedure
Using a centimeter measuring tape, determine the following dimensions and record them in the table on the
worksheet.
Circumference of the chest at the level of the 3rd
A persons normal respiratory rate is determined by
rib (just under the armpit) during resting and
the medullary rhythmicity center in the medulla oblonforced inhalation and exhalation (4 measuregata of the brain. This area of the brain in turn rements).
ceives input from chemoreceptors in the body which
Circumference of the abdomen at the level of the
are sensitive to carbon dioxide levels (actually sensiumbilicus during resting and forced inhalation
tive to H+ levels) in the blood. There are chemoreand exhalation (4 measurements).
ceptors in the aortic arch and carotid bodies (the peripheral chemoreceptors) as well as chemoreceptors in Using the calipers, determine the following dimenthe medulla (central chemoreceptors). The peripheral sions and record them in the table in the worksheet.
chemoreceptors can be influenced by any source of H+ Anterior-posterior thickness of the chest at the
in the blood whether from changes in CO2 level
level of the 3rd rib during resting and forced inha(because CO2 is converted to H+ and bicarbonate by
lation and exhalation (4 measurements). Position
carbonic anhydrase) or changes in acid production (i.e.
the calipers over the shoulder to make these measlactic acid). The central chemoreceptors are only senurements.
sitive to changes in respiratory CO2 levels because H+ Side-to-side thickness of the chest at the level of
cant cross the blood brain barrier (CO2 does cross and
the 3rd rib during resting and forced inhalation
is converted into H+ and bicarbonate by carbonic anand exhalation (4 measurements).
hydrase). At rest, alterations in ventilation pattern will
cause alterations in blood carbon dioxide levels. Thus, Measuring Lung Volumes
increased respiratory rate (hyperventilation) will decrease blood CO2 while decreased respiratory rate
Procedure
(hypoventilation) will increase blood CO2. Such alterations in blood CO2 will, in turn, induce changes in the See figure 2 for spirometer setup, turn on the IWorx
respiratory rate and depth in order to bring blood CO2 unit, then start up the LabScribe software and from the
levels back to normal.
settings menu choose load and 11 Mechanisms of
Breathing then Settings again and choose MechaAlthough it is true that changing blood carbon dioxide nisms of Breathing. The resulting screen should have
level will alter the respiratory rhythm, it should be
2 recording screens labeled Air Flow and Volume.
noted that exercise induced changes in the respiratory When you breathe through the spirometer the air flow
rhythm are NOT due to changes in blood carbon diox- is detected by the iWorx air flow transducer and the
ide level. During exercise the autonomic nervous system immediately enters a state of sympathetic tone
(sympathetic dominance) and the increased activity of
the noradrenergic neurons and blood levels of epinephrine serve to increase the respiratory rhythm prior
to any actual change in blood carbon dioxide level.
In todays laboratory we will be examining each of
these physiological concepts: 1) How thoracic and
abdominal dimensions change with breathing, 2) Lung
volumes before and after exercise and 3) How changFigure 2. iWorx Spirometer
Revised Fall 2014
Page 90

ume (in liters) of 5 respiratory cycles by
positioning the left cursor on the trough
Do not measure from this screen!
and the right cursor on the peak of each of
5 different cycles. Record the average of
these 5 tidal volumes in the worksheet and
time
your journal. Measure the peak to peak
volume distance (time) for 5 respiratory cycles.
Divide 300 by this number to get the resTake all measurements from this screen
piratory rate in breaths per minute (the
same way you measured heart rate). Record these data in your journal. From the
respiratory rate and the tidal volume you
can determine how much air your subject
breathes every minute. This is called the
respiratory minute volume and is calculated by multiplying the respiratory rate
Figure 3. Main spirometry screen with tidal volumes shown.
by the tidal volume. Record these values in the
journal and in your worksheet.
software converts
VERY IMPORTANT
this into air flow in
Expiratory Reserve Volume (ERV)
Do not breathe through the flow head liters. The bottom
during the first 5 seconds after clicktracing automatically
The subjects nose should be closed off, as previously
ing on the Record button. The iWorx converts the rate of
described. To determine the expiratory reserve voluses this time to zero the volume reair flow into a volume, which is the additional amount of air that can be
cording. If your baseline is not level
ume. You must alyou probably did not allow enough
low the equipment to expelled beyond normal exhalation, the subject should
time before breathing into the flow
warm up for at least breathe normally for a few breaths, then exhale maximally, without looking at the computer screen. The
head.
10 minutes before
expiratory reserve volume (ERV) is measured as the
beginning the exerincrease in exhaled air volume over and above a norcises and you should NOT breathe through the machine after commencing recording for about 5 seconds mal exhalation (see Figure 4). Repeat this measurement 3 times and calculate the average. Record these
before placing your mouth on the mouthpiece. In
addition, always breathe through the spirometer head screens and calculations in your journal and worksheet.
with the tubing pointing up to avoid the formation of
condensation in the tubing. While inhaling you should
Vital Capacity (VC)
see the tracing on the computer screen rise. If it decreases you should breath through the other end of the
The nose should be closed off as before. To determine
spirometer.
the vital capacity (the maximum volume of air the
subject can exchange), the subject should, after a few
Tidal Volume (TV)
normal breaths, inhale as hard as possible, then exhale
Have your subject put on a nose clip (or hold the nose into the spirometer as deeply as possible while recording, without looking at the computer screen (see Figclosed with their fingers). About 5 seconds after
clicking the Record button have
the subject breathe as normally as
possible for about 10 cycles (see
Figure 3). The subject should
TV
NOT be watching the recorder.
Copy the resulting screen into
ERV
your journal after you enter the
heading into the journal.
Using the lower Volume screen,
determine the average tidal volFigure 4. Measuring ERV
Revised Fall 2014
Page 91
VC
Figure 5. Measuring Vital Capacity

ure 5). The greatest volume
reached is a measure of the vital
capacity. Repeat this 3 times and
calculate the average. Record
these data in the journal and worksheet.
The determination of residual volume

in humans requires specialized equipment and the use of inert radioactive
tracers. This determination is beyond
the scope of an undergraduate physiology laboratory. Thus we assume a
residual volume of 1.2 liters.
minute volume. Enter these data in

the worksheet and your journal.
In the next two exercises you will
examine how blood carbon dioxide
level influences the respiratory
control centers in the brain. Since
we cant directly alter carbon dioxide levels we are going to indirectly change the levels and examine
Inspiratory Reserve Volume (IRV)
the consequences.
From Figure 1, you can see that:
Changing Carbon Dioxide Levels
VC = TV + ERV + IRV
With measurements for VC, TV and ERV, you can
solve for IRV and calculate the inspiratory reserve volume of your subject. Record this value in the journal.
Total Lung Capacity (TLC)
From Figure 1 you can also see that:
TLC = VC + RV
Decreasing Blood CO2 (Hyperventilation)

Hyperventilation will decrease carbon dioxide levels
in the blood. Since carbon dioxide level is a proximal
stimulator of breathing, if we hyperventilate for a period of time we will release more CO2 from the blood
and we should be able to hold our breath longer than if
we didnt hyperventilate. You can change subjects for
this exercise if you wish and you do not need to use
the spirometer.
Assuming that the average human has a residual volume First, the subject should hold his/her breath for as long
as possible with a lab partner timing him/her. This
of 1.2 liters, solve this equation for TLC. Record this
time should be recorded on the worksheet.
value on the worksheet.
Then, the same subject should now hyperventilate by
breathing deeply at the rate of 2 breaths/second for
about 30 seconds; then the subject should take a deep
breath and hold it as long as possible. Record this
Have the same subject exercise moderately for 2
minutes. Immediately have the subject close off his/her time on the worksheet.
nose, and then record his/her breathing in order to comIncreasing Blood CO2 (Hypoventilation
pare rate and depth with the resting pattern previously
recorded. Copy this tracing into the journal. Determine
For this exercise you will again use the spirometer and
the post exercise TV, respiratory rate and respiratory
Effects of Exercise
Revised Fall 2014
Page 92

recorder. The same volunteer you used for the spirometry previously should record at least 5 normal breathing
cycles (mark the recording pre-CO2). They should
then breathe into a paper bag for 3 minutes (or as long
as possible). The volunteer should then record their
breathing in the same fashion as before (mark the recording post-CO2). Measure the tidal volume and
respiratory rate before and after the volunteer breathed
into the paper bag. Copy and record these data into the
journal and worksheet.
Revised Fall 2014
Page 93
Revised Fall 2014
Page 94
Journal: __________


Date:
Name:
Section:
Thoracic and Abdominal Dimensions

1) Enter the chest and abdominal measurements in the following table.
Resting
Inhalation
Exhalation
Forced
Inhalation
Exhalation
Chest
Circumference (cm)
Abdominal
Circumference (cm)
Ant. - Post. Chest
Dimension (cm)
Side-to-Side Chest
Dimension (cm)
2) Do these measurements correspond with your understanding of how changes in thoracic and abdominal cavity
dimensions should change during breathing? Please explain.
__________________________________________________________________________________________
__________________________________________________________________________________________
__________________________________________________________________________________________
3) Describe how the diaphragm moves/works during a normal breathing cycle.
__________________________________________________________________________________________
__________________________________________________________________________________________
__________________________________________________________________________________________
__________________________________________________________________________________________
4) What happens to intra-alveolar pressure during inhalation and exhalation?
__________________________________________________________________________________________
__________________________________________________________________________________________
__________________________________________________________________________________________
Revised Fall 2014
Page 95
Name:
(continued)
5) How do abdominal muscles contribute to inhalation and exhalation? Explain.

__________________________________________________________________________________________
__________________________________________________________________________________________
__________________________________________________________________________________________
__________________________________________________________________________________________
6) During forced breathing, you might expect the abdominal circumference to change dramatically. Why?
__________________________________________________________________________________________
__________________________________________________________________________________________
__________________________________________________________________________________________
__________________________________________________________________________________________
Lung Volumes and Exercise

Pre-exercise Tidal Volume (TV), Respiratory Rate and Respiratory Minute Volume
Tidal volume
(liters)
Respiratory Rate:
Time for 5 cycles = ____________ sec
Respiratory rate (300/time for 5 cycles) = __________ breaths/min
Average tidal volume = __________ Liters
Average:
Respiratory minute volume (TV X Respiratory rate) = __________ Liters/min
7) Record these data in the summary table on the next page.

Pre-exercise Expiratory Reserve Volume (ERV)
ERV (liters)
Average:
8) Enter the ERV value in the summary table on the next page.
Revised Fall 2014
Page 96
Name:
(continued)
Vital Capacity (VC)
Summary Data
VC (liters)
Parameter
Pre-Exercise
Post-Exercise
Difference
Tidal volume
(L)
Average:
9) Enter the average VC into the summary table.
IRV = VC - (TV + ERV)
IRV = __________ liters
10) Enter the IRV into the summary table.
Total Lung Capacity (TLC)
TLC = VC + RV
(RV = residual volume = 1.2 liters)
Respiratory
rate (breaths/
min)
Respiratory
minute volume
(L/min)
Expiratory reserve volume
(L)
Inspiratory
reserve volume
(L)
Vital Capacity
(L)
Total Lung
Capacity (L)
TLC = _________ liters

11) Enter the TLC into the summary table.
Effects of Exercise on Tidal Volume, Respiratory Rate and Respiratory Minute Volume
Post-Exercise Tidal
Volumes (liters)
Respiratory Rate:
Time for 5 respiratory cycles = _________ sec
Average post-exercise TV = __________ Liters
Average:
Revised Fall 2014
Post-exercise Respiratory Minute Volume (TV X Respiratory rate) = __________ Liters/min
Page 97

Name:
Name: _______________
(continued)
12) From the summary table it should be evident that respiratory minute volume changes most dramatically with exercise. What is the physiological significance of this change?
________________________________________________________________________________________________
________________________________________________________________________________________________
________________________________________________________________________________________________
________________________________________________________________________________________________
Changing Carbon Dioxide Levels

Effects of Hyperventilation (decreased blood CO2)
13) Before hyperventilating subject held breath for _________
14) After hyperventilating subject held breath for _________
15) Can you think of a sport in which this might be an important (and legal) part of increasing performance?
__________________________________________________________________________
Pre Hypoventilation Tidal volume
Respiratory rate:
Pre Hypoventilation
Tidal Volume
(liters)
Time for 5 respiratory cycles = __________ sec

Average tidal volume = _________ liters
Respiratory minute volume = __________ liters/min
Post Hypoventilation (increased blood CO2) Tidal Volume
Respiratory rate:
Average:
Post Hypoventilation Tidal Volume
(liters)
Time for 5 respiratory cycles = __________ sec

Average tidal volume = _________ liters
Respiratory minute volume = __________ liters/min
Average:
Revised Fall 2014
Page 98

Name:
Name: _______________
(continued)
16) What differences in the tidal volume and/or respiratory rate can you see from the beginning to the end of the 3 minute hypoventilation period?
__________________________________________________________________________________________
__________________________________________________________________________________________
__________________________________________________________________________________________
__________________________________________________________________________________________
17) What effect did hypoventilation have on respiratory minute volume? Why?
__________________________________________________________________________________________
__________________________________________________________________________________________
__________________________________________________________________________________________
__________________________________________________________________________________________
18) How is blood PCO2 detected in the human body?
__________________________________________________________________________________________
__________________________________________________________________________________________
__________________________________________________________________________________________
__________________________________________________________________________________________
__________________________________________________________________________________________
19) Where are the most important receptors and how do they work?
__________________________________________________________________________________________
__________________________________________________________________________________________
__________________________________________________________________________________________
__________________________________________________________________________________________
__________________________________________________________________________________________
Revised Fall 2014
Page 99

Name:
Name: _______________
(continued)
Revised Fall 2014
Page 100
Journal Format for Mechanisms of Breathing
(cont.)
(Guide to producing
Name:journal)
_______________
a complete
Your Name
Lab Partners names
Laboratory Section
Date
A. PRE-EXERCISE TIDAL VOLUME AND RESPIRATORY RATE

Paste a representative screen showing
the measurement of tidal volume.
Pre-exercise average (from the summary table) tidal volume = _________ Liters
Paste a screen showing the measurement of pre-exercise respiratory rate.
Pre-exercise respiratory rate = __________ breaths/min
B. EXPIRATORY RESERVE VOLUME
the measurement of expiratory reserve
volume.
< (exhale)
Average expiratory reserve volume (from summary table) = __________ Liters

C. VITAL CAPACITY
(deep inhale) >

the measurement of vital capacity.
< (deep exhale)
Average vital capacity (from summary table) = __________ Liters

E. EFFECTS OF EXERCISE
the measurement of post-exercise tidal
volume.
Average post-exercise tidal volume (from summary table) = __________ Liters
Paste a screen showing the measurement of post-exercise respiratory rate.
Post-exercise respiratory rate = __________ breaths/min
Revised Fall 2014
Page 101
Journal Format for Mechanisms of Breathing

(continued)
F. EFFECTS OF HYPOVENTILATION
the measurement of prehypoventilation tidal volume.
Average pre-hypoventilaion tidal volume = __________ Liters
Paste a screen showing the measurement of pre-hypoventilation respiratory
rate.
Pre-hypoventilation respiratory rate = __________ breaths/min
the measurement of posthypoventilation tidal volume.
Average post-hypoventilaion tidal volume = __________ Liters
Paste a screen showing the measurement of post-hypoventilation respiratory rate.
Post-hypoventilation respiratory rate = __________ breaths/min
Make sure all 10 journal pages are in order and turn them in with the worksheet
Revised Fall 2014
Page 102
Biology 335 Human Physiology: Lung Disease
Laboratory #12
Restrictive and Obstructive Lung Disease
using the iWorx spirometry system.
Forced exhalation is utilized for this measurement
1. Gain an understanding of the importance of air
because it is more sensitive to airway changes than
flow during inhalation and exhalation
inhalation. During exhalation the lungs and airways
2. Simulate the effects of restrictive lung disease on are subjected to a positive pressure which tends to
air flow during respiration.
force the airways to partially close. If the airways are
3. Simulate the effects of obstructive lung disease on already narrower than normal this will result in a
air flow during respiration.
measurable decrease in airflow. On the other hand,
during inhalation, the lungs and airways are subjected
to a negative pressure which forces airways open thus
Air Flow During Respiration
masking abnormal airway issues. The FEV1/FVC is
usually expressed as a percentage (simply multiply
FEV1/FVC by 100) and normal values are 80% or
In the last laboratory (Chapter 11), spirometry was
higher.
used to measure the major lung volumes and capacities during human respiration. In addition, the regulation of the respiratory rhythm was explored by manipNormal FEV1/FVC
ulating blood CO2 levels through hyperventilation or
hypoventilation.
Procedure
Human pulmonary diseases are often diagnosed by
measuring lung volumes and the rate of air flow
Set up the spirometer as describe in laboratory #11.
through the pulmonary airways. Pulmonary diseases
From the settings menu choose load and 12 Lung
are usually classified as either restrictive or obstrucDisease then Settings again and choose Lung Disease.
tive. Restrictive diseases are those characterized by a The resulting screen should have 2 recording screens
decreased ability of the lungs to change volume result- labeled Air Flow and Volume.
ing in a decrease in the vital capacity and an increase
in residual volume. Any disease which decreases lung With the computer recorder on, the volunteer should
compliance (emphysema) or decreases the ability of
breathe normally for a couple of cycles then inhale
the lung to be inflated (myasthenia gravis) is considmaximally, hold their breath momentarily, then exhale
ered a restrictive disease. Obstructive diseases are
maximally AS LONG, FAST AND HARD AS POScharacterized by reduced air flow through the pulmo- SIBLE (keep that nose closed!). Repeat this for a total
nary airways (emphysema or asthma).
1 second
The determination of lung volumes can
be carried out using spirometry as in Laboratory #11. These measurements can
be used to detect changes in lung inflation in the diagnosis of restrictive lung
diseases. The determination of changes
FEV1
in airflow, however; requires a different
FVC
measurement. The rate of air flowing
through the pulmonary airways is measured by recording a patients FEV1/FVC.
FEV1 is the Forced Expiratory Volume in
the first second of the exhalation. FVC
is the Forced Vital Capacity or the total
volume exhaled during the same exhalaFigure 1. FEV1/FVC
tion. Figure1 illustrates how FEV1/FVC is measured
Objectives
Page 103
Revised Fall 2014

This laboratory exercise was adapted from a protocol developed by Dr. Debra Mulliken-Kilpatrick of Boston College.

of 3 trials, allowing some recovery time between trials. This is very similar to the measurement of VC
performed in Laboratory #11, however, if the subject
pauses momentarily prior to exhaling it will make the
measurement easier (see Figure 1).
Using the double cursors (use the arrow keys on the
keyboard for precision), measure the TV, IRV and
ERV as shown in Figure 2. Then from the beginning
of the exhalation, mark off 1 second (T2-T1) and
determine the volume exhaled during that 1 second
(V2-V1; Forced Expiratory Volume in the first second[FEV1]). Then move the right hand cursor over to
the lowest point of exhalation (Forced Vital Capacity;
FVC) and determine the volume (V2-V1). Divide the
FEV1 by the FVC then multiply by 100 to express the
volume as a percent of the maximal exhalation. This
is called the FEV1/FVC. Repeat this measurement 2
more times and enter the data in the table provided in
the worksheet. Copy your best tracing into the journal
and record the average FEV1/FVC in the worksheet
and the journal.
Restrictive Disease
Restrictive pulmonary diseases are characterized by a
decrease in lung compliance or ability to expand the
thoracic cavity. This being true, we can induce a restrictive condition by having a subject wear a medical
corset which decreases the maximum expansion of the
thoracic cavity.
A volunteer in your lab group should put the medical
corset on tight enough to restrict expansion without
being unduly uncomfortable. Now
have the volunteer
breath through the
spirometer as described in the previous exercise. Record three normal
tidal volumes followed by a maximum inhalation
(pause) then a maximal exhalation
(long, hard and
forceful). See Figure 2 for an example. If your output
doesnt appear similar to the example
you will need to ask
Average Adult Lung Volumes

Lung Volume
Volume (ml)
Tidal volume (TV)
500
3100
Expiratory Reserve Volume (ERV)
1200
Forced Vital Capacity (FVC)
4800
Residual Volume (RV)
1200
your instructor for assistance.

Repeat the measurements 2 more times with the same
volunteer resting briefly between each measurement (3
times total) Record the data in the table in the worksheet and provide a sample recording in the journal. If
you dont see a change in the FVC you may need to
make the corset a little tighter.
Note: The corset should be tight but not uncomfortable. However, it should be tight enough to restrict
the expansion of the chest to some degree.
Obstructive DiseaseDecreased Airway

Diameter
Obstructive pulmonary diseases are characterized by
decreases in air flow through the respiratory tree. We
can examine the impact of changes in the pulmonary
airways in the laboratory by decreasing the diameter
Figure 2. Pulmonary Function Testing

Revised Fall 2014
Page 104

of the tube leading to the spirometer head.
Use the narrowed airway attachment on the spirometer
head to test this.
Measure your volunteers pulmonary function volumes
as you did in the previous exercises. Repeat 2 more
times for a total of 3 sets of measurements. Enter your
data in the table provided in the worksheet and include
a representative recording in your journal.
Revised Fall 2014
Page 105
Journal: __________


Date:
Name:
Section:
Normal FEV1/FVC
1. Enter the normal FEV1, FVC and FEV1/FVC data in the table provided below. When complete, enter the average TV, IRV, ERV, FVC and FEV1/FVC in the summary table.
TV (ml)
IRV (ml)
ERV (ml)
FEV1 (Liters)
FVC (Liters)
FEV1/FVC
(%)
Averages:
2.Why is the measurement of FEV1/FVC used instead of the rate of inhalation in pulmonary function testing?
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
3. What affect will a poor seal around the mouthpiece have on the results of an FEV1/FVC measurement?
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
Restricted and Obstructive Pulmonary Diseases

4.Enter the data collected from the subject wearing the medical corset into the table provided here. When complete, enter the average TV, IRV, ERV, FVC and FEV1/FVC in the summary table.
TV (ml)
IRV (ml)
ERV (ml)
FEV1 (Liters)
FVC (Liters)
FEV1/FVC
(%)
Averages:
Revised Fall 2014
Page 106

5. Enter the data collected from the volunteer breathing through the obstructed spirometer. When complete, enter
the TV, IRV, ERV, FVC and FEV1/FVC in the summary table.
TV (ml)
IRV (ml)
ERV (ml)
FEV1 (Liters)
FVC (Liters)
FEV1/FVC
(%)
Averages:
Summary Table
Measurement
Restrictive
Obstructive
TV (ml)
IRV (ml)
ERV (ml)
FVC (L)
FEV1/FVC (%)
6. Why is the rate of exhalation a better measurement than the rate of inhalation for the diagnosis of obstructive
diseases?
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
7. Do your results agree with the expected physiological changes for restrictive and obstructive pulmonary diseases? Why or why not?
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
Revised Fall 2014
Page 107

8. What changes in the FEV1/FVC would be expected with restricted pulmonary disease?
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
9. What changes in FEV1/FVC should you see with obstructive disease?
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
10. What pulmonary volume should be most altered by restricted lung disease? Does your data support this expectation?
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
11. What effect should restrictive disease have on the residual volume? Why?
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
12. Why is asthma considered an obstructive disease?
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
13. Why is epinephrine an effective emergency treatment for an acute asthmatic attack?
___________________________________________________________________________________________
___________________________________________________________________________________________
___________________________________________________________________________________________
Revised Fall 2014
Page 108
Biology 335 Human Physiology: Lung Diseases
Journal Format for Lung Diseases
(cont.)
(Guide to producing
Name:journal)
_______________
a complete
Your Name
Lab Partners names
Laboratory Section
Date
A. NORMAL PULMONARY FUNCTION VOLUMES

the measurement of normal pulmonary
function. See Figure 2 for example
B. RESTRICTIVE DISEASE
D. FEV AND FORCED VITAL
Paste a representative1 screen showing
CAPACITY
the measurement of pulmonary function while wearing the medical corset.
C. OBSTRUCTIVE DISEASE DECREASED AIRWAY DIAMETER

the measurement of pulmonaryAverage
func- FEV1/
tion while breathing
narrowed
device.
FVC (from summary table) =
__________ %
Revised Fall 2014
Page 109
Revised Fall 2014
Page 110
Biology 335 Human Physiology: Metabolic Rate
Laboratory #13
Basal Metabolic Rate
Objective
1. To determine the mean metabolic rate of student
the conditions listed above might be a problem. Instead, a very handy way to estimate metabolic rate
is to determine oxygen consumption (since anaerobic metabolism contributes very little to total metabolism in mammals).
volunteers.
2. Compare human metabolic rate to the metabolic
rate of a small mammal.
Most body heat is produced by the physiological

oxidation of fats, proteins and carbohydrates.
These foodstuffs are digested in the gastrointestinal tract, and are absorbed into the vascular system
(blood and lymph) as fatty acids and glycerol,
Background
amino acids, and simple sugars. Each type of
foodstuff, when oxidized, will
Since the time of Lavoisier (1743-1794), it has
1 kcal (large calorie) is
liberate a given amount of
the amount of heat rebeen realized that the body behaves somewhat
heat for each liter of oxygen
quired to warm 1 kilogram
like a furnace, requiring fuel to burn as a source
consumed. The amount of
of water by 1 degree Celof energy with a consequent elimination of heat.
heat liberated is called the
sius.
The chemical reactions that occur in the body may
calorific value of a liter of oxbe classified as energy trapping (endergonic) or energy
ygen for that material. The established values are:
releasing (exergonic). The exergonic reactions make
available the largest increments of energy for four eskcal per L O2
sential processes: (1) energy for muscle movement; (2)
Fat
4.69
energy for digestion and associated processes; (3) enerProtein
4.48
gy for adjustment of body temperature; and (4) energy
Carbohydrates
5.00
for basal maintenance of all the body cells. This last is
Typical Mixed Diet
4.83
basal metabolism, and in order to measure it the first
three energy needs must be held to a minimum. Basal
Although there are specific calorific values for each
metabolic rate (BMR) as determined clinically is the
foodstuff, the average person is on a mixed diet
metabolic level of the individual under the following
which has been found to have a calorific value very
three conditions corresponding to minimal levels for the
close to 4.83 kcal (large calories) per liter. Thus,
first three of the above considerations:
for each liter of oxygen consumed, 4.83 large calories of heat are produced.
1. Mentally relaxed, rested in the recumbent position for 30 minutes before the test, and in a reOnce oxygen consumption has been determined,
cumbent position during the test.
this can be related to the total average number of
2. In a post-absorptive state; that is, the subject
kilocalories (kcal) consumed for a given amount of
has not taken any food for 12 hours prior to the
oxygen consumed (4.83 kcal/L oxygen). This will
test.
give you an equivalent to the rate of heat loss per
3. At a comfortable environmental temperature
given period of time. This can be compared to
between 20o to 25o C (68o to 77o F) and at a
standard population averages for a given body size
normal body temperature. This is known as
and expressed as a percentage of that value.
the thermal neutral temperature.
There are a number of methods for measuring metabolic
rate including heat production which is a waste product
of cellular metabolism (see formula below). If you
think about this a few minutes you will realize that determining total body heat production over time under
Revised Fall 2014
Page 111

There are several standard ways which this information can be expressed. The one we shall use is the
Mayo Foundation Normal Standard. The standard is
expressed as large calories (kcal) of heat produced per
square meter of body surface area per hour. In computing the metabolic rate of the given individual
(which is compared with that of the standard), it is
necessary to know his/her body surface area. The
body surface area is read off the DuBois Surface Area
Chart when height and weight of the subject are
known.
The basal metabolic rate (BMR) is expressed as the
percentage amount by which the metabolic rate of the
individual lies above or below the standard. Thus, if
any individual had a metabolic rate of 40 calories per
hour per square meter of body surface area and his
standard was 30 calories per hour per square meter of
body surface area, his metabolic rate would be 40/30
of the standard, or 33.3 percent above the standard.
Thus, his BMR would be expressed as +33.3.
Why use surface area? It turns out that an animals
(or a persons) metabolic rate is affected by body surface area. What will lose heat faster, a 10 cm3 steel
block or a 1 cm3 steel block? The smaller object will
lose heat much faster than the large block. This is
because the smaller object actually has a larger surface area for its given mass. Generally, smaller animals exhibit higher metabolic rates on a per gram
basis. The total heat production for a human is much
greater than that for a mouse, but based on a mouses
body weight they exhibit higher metabolic rates than
humans.
The relationship between basal metabolic rate and
body size for many different animals is illustrated in
the next two figures. The first graph shows that as
body size increases, heat production per day increases.
Mass-specific
BMR
mass (g)
We are going to determine and compare the average
BMR of a group of students and the average BMR of a
group of laboratory mice.
Human Basal Metabolic Rate

Procedure
Materials:
MedGem Indirect Calorimeter
Nose clip
Calculator
Subject:
It will be impossible to measure oxygen consumption
under truly basal conditions in our laboratory. There is
simply too much going on in the room. However, at least
6 students in each class should come to class without
eating or drinking caffeinated beverages. The reason for
this is that they will be in a post-absorptive state, satisfying at least one of the conditions under which BMR
should be measured.
Oxygen consumption of the subjects will be determined
using a MedGem Indirect Calorimeter. Each subject will
take turns determining his or her oxygen consumption
according to the directions supplied by your instructor.
Log[kcal/day]
Log [mass] (kg)

Revised Fall 2014
The next figure shows that as animals become larger, the

mass-specific BMR actually decreases. The massspecific BMR is an animals basal metabolic rate expressed on a per gram body weight basis. In other words,
it is the metabolic rate for each gram of mass of an animal.
The MedGem
The MedGem is a state-of-the-art, handheld, indirect calorimeter
that accurately measures oxygen consumption (VO2). Indirect
calorimetry is a process whereby the rate of energy expenditure
is estimated based upon the rate of oxygen consumption or carbon dioxide production.
Page 112

the standard BMR for a person of the same sex and
age as your subject.
11. Basal metabolic rate. BMR can be expressed as a
The numbers below refer to specific lines on the lacomparison to this published standard. The number
boratory worksheet. You can enter your numbers on
on line (9) may be above or below the standard. Exthat sheet as you progress through the calculations.
press the metabolic rate as a percentage of the standard. Divide line (9) by line (10), then multiply by
1. Observed volume of oxy100. A percentage greater
THE MAYO FOUNDATION NORMAL
gen consumed per minute
than 100 means your value
STANDARDS
(mL/min). This value will
was above the standard; a perMean Basal Metabolic Rate
be taken from the indirect
(Kcal per square meter per hour) centage less than 100 means
calorimeter as directed by
Age
Males
Females
your value was below the
17
44.80
41.45
the instructor.
standard.
17.5
44.03
40.74
2. Volume in liters per mi12. Mass-specific metabolic
18
43.25
40.10
nute. Convert the oxygen
18.5
42.70
39.40
rate. Another way to express
consumption value on line
19
42.32
38.85
BMR is in units of heat pro19.5
42.00
38.30
(1) to liters per minute by
duced per hour per gram body
20-21
41.13
37.82
dividing by 1000.
mass. To calculate this, di22-23
40.82
37.40
3. Heat production per mi24-27
40.24
36.74
vide the number on line (4) by
nute in kcal. Multiply the
28-29
39.81
36.18
the subjects mass in grams
30-34
39.34
35.70
number on line (2) by 4.83.
on line (7). Report this value
35-39
38.68
34.94
The reason for using this
in the worksheet.
40-44
38.00
33.96
figure is discussed in the
Introduction. Briefly, you
are taking your subjects oxygen consumption
Record your group data in the table on the blackboard,
(liters) and multiplying it by 4.83 kcal/liter of
overhead or instructors computer as directed by your inoxygen, which is an estimate of how much heat
structor. BMR data, surface area and body weights
is produced for each liter of oxygen a person
should be recorded for subsequent calculation of class
consumes. This converts units of oxygen conaverages.
sumption into units of energy (kcal).
4. Heat production per hour. Multiply the numCalculate the class average human surface area to mass
ber on line (3) by 60. This is a very meaningful
ratio by dividing the class average surface area (m2) by
number because it reflects the rate at which the
the class average mass (g). Record this value in the
subject burns calories (which are actually kiloworksheet.
calories) while sitting quietly.
5. Subjects height. Express this in cm (1 in =
2.54 cm).
Mouse Resting Metabolic Rate
6. Subjects mass. Convert pounds to kilograms
by dividing by 2.2.
7. Subjects mass. Convert kilograms to grams by
multiplying by 1000.
In this exercise we will determine the average BMR for
8. Subjects body surface area. Use the following
a group of mice.
formula to determine the body surface area:
Calculations:
Procedure
..
height (cm) X mass (kg)
S A (m )
9.
3600
Heat production per hour per square meter

of body surface. Divide line (4) by line (8).
This is one way to express the metabolic rate of
your subject. It must now be compared with
published standard values.
10. Standard. Refer to the table of Mayo Foundation Normal Standards in this handout and find
Revised Fall 2014
Materials:
Metabolism chambers
Soap solution
Balance
Barometer
Thermometer
Page 113

The Mouse Metabolic Chambers
Subjects:
Adult laboratory mice.
As we did for human subjects earlier, we will calculate metabolic rates for mice using oxygen consumption data. Each
group of students will obtain a mouse, determine its mass,
and place it in a metabolism chamber. The metabolism
chamber is a cylinder with a large one-hole rubber stopper
blocking one end. Soda lime has
been place in the bottom of the chamber to absorb carbon dioxide. To isolate the mouse from the soda lime and to limit
its movement during the metabolic rate determination it will
be placed in a small mesh cylinder which is sealed at either
end with a cap. When a mouse is placed in the mesh cylinder
and the metabolism chamber is closed, the carbon dioxide
which the mouse generates via cellular respiration will be absorbed by the soda lime. The resulting decrease in volume
represents oxygen consumption. The rubber stopper has a
graduated tube penetrating it such that if you place a soap
bubble at the end of the tube, when the mouse breathes and
the carbon dioxide is absorbed, the volume of air in the cylinder decreases and the soap bubble will be sucked in towards the mouse. The actual oxygen consumption which
you measure needs to be multiplied by a correction factor
which adjusts for barometric pressure and altitude such that
all measurements are standardized to these conditions. You
didnt have to do this for the human data because the
MedGem automatically introduces this correction.
The mouse is placed in a tube containing soda lime

which absorbs any carbon dioxide the mouse produces. By placing a soap bubble at the end of the pipette, the chamber becomes a closed space. As the
mouse consumes oxygen and exhales carbon dioxide, the carbon dioxide is absorbed and the volume of
gas in the chamber decreases in an amount equivalent to the amount of oxygen the mouse consumed.
The decreased volume pulls the soap bubble through
the pipette. This change in volume represents the
oxygen consumption of the mouse.
Calculations:
1.
2.
3.
4.
Oxygen consumption per minute (mL/min). This value is the average oxygen consumption of your mouse.
Chamber temperature (oC). The temperature in the
metabolic chambers as provided by the instructor.
Barometric pressure (mm Hg). Current barometric
pressure as supplied by the instructor.
Correction factor (from table or calculated). This factor standardizes your oxygen consumption for variations
in barometric pressure and temperature.
Correction Factor
Baro. P. (mm Hg)

760
1.
2.
3.
4.
5.
6.
7.
8.
9.
Weigh the mouse using the supplied balance.

Place the mouse in the metabolic chamber.
Allow the mouse to acclimate (get used to) its surroundings for at least 5 minutes.
Insuring that the chamber is closed, place a soap bubble
over the end of the pipette using a moistened finger tip.
Record the oxygen consumption over a given period of
time. Try to record the time it takes for a full 5 mL of
oxygen to be consumed. If this proves very difficult record both the volume (at least 2 mL) and the time and determine the oxygen consumption in mL/min based upon
these data.
Repeat this 5 additional times. Between trials, briefly
flush the air in the chamber by removing the rubber
stopper and allowing room air to enter.
Record your data on the laboratory worksheet.
Determine the average oxygen consumption of your
mouse based on the six measurements you obtained.
Use the oxygen consumption value for your mouse to
calculate the various BMR measurements (see calculations below).
Revised Fall 2014
273
Temp. (oC) + 273
5.
Corrected oxygen consumption for the mice.

Multiply line 1 by line 4.
6. Oxygen consumption in L/min. Convert to L/min by
dividing line 5 by 1000.
7. Heat production (kcal/min). Multiply line 6 by 4.83.
8. Heat production (kcal/hour). Multiply line 7 by 60.
9. Mouse mass (g).
10. Surface area (m2). Use the formula in the box below to
calculate your class average mouse surface area.
11. Heat production (kcal/h/m2). Divide line 8 by line 10.
12. Mass specific BMR (kcal/h/g). Divide line 8 by line 9.
Formula for estimating rodent surface area:

S.A. (cm2) = 0.437+(2.143 X mass(g))
Divide this by 10,000 to give m2
Page 114

Record your group data in the table on the blackboard, overhead or instructors computer as directed by your instructor. BMR data, surface area
and body weights should be recorded for subsequent calculation of class averages.
Surface/Mass Ratios
Calculate the surface area to mass ratio by dividing the class average surface area (m2) by the
body weight (g). Record this value in the worksheet.
The surface to mass ratio of a species or organism
represents the proportion of that organisms mass
which is exposed to the environment. An animal
with a larger surface area to mass ratio will lose
heat from their bodies more rapidly. This heat
must be replaced by metabolic activity. Smaller
animals will exhibit a larger surface area to mass
ratio and will thus typically exhibit higher per
gram metabolic rates.
Revised Fall 2014
Cautions:
1. Mice may bite. If you are not used to
handling mice, allow the instructor to
assist you in transferring the mice from
their cages to the chamber and back
again.
3. Return each mouse to the same cage
from which it was taken. This will
minimize disruption of their social order
and prevent unnecessary fighting among
the animals.
Page 115
Revised Fall 2014
Page 116
Journal: __________


Date:
Name:
Section:
Human Basal Metabolic Rate

1) Enter your group volunteers data in the spaces provided below:
Subjects name: ________________________
Subjects sex: _______
Subjects age: _______
1-
Oxygen consumption (mL/min)
______
2-
O2 volume in L/min
______
3-
Heat production (kcal/min)
______
4-
Heat production per hour (kcal/hr)
______
5-
Subjects height (cm)
______
6-
Subjects mass (kg)
______
7-
Subjects mass (g)
______
(m2)
8-
Surface area
9-
Heat production per hour per m2
______
______
10- Standard BMR
______
11- BMR (% of standard)
______
12- Mass specific BMR (kcal/h/g)
______
THE MAYO FOUNDATION NORMAL

STANDARDS
Age
17
17.5
18
18.5
19
19.5
20-21
22-23
24-27
28-29
30-34
35-39
40-44
Mean Basal Metabolic Rate

(kcal per square meter per hour)
Males
Females
44.80
41.45
44.03
40.74
43.25
40.10
42.70
39.40
42.32
38.85
42.00
38.30
41.13
37.82
40.82
37.40
40.24
36.74
39.81
36.18
39.34
35.70
38.68
34.94
38.00
33.96
2) Enter the human volunteer data for each group in the table below and calculate the means:
Class Data Human

Lab. Group
Heat Production
(kcal/h)
Mass (g)
Surface Area
(m2)
Heat Production (kcal/h/m2)
Mass Specific
BMR (kcal/h/g)
1
2
3
4
5
6
Class
Average
Revised Fall 2014
Page 117

Date:
Name:
Section:
3) Calculate the average human surface area to mass ratio using your class data:
Class Average Human Surface Area to Mass Ratio:
________________________________________
Do the math! Surface area divided by mass.
4) Under what circumstances might the BMR data you obtained be inaccurate or a poor representation of a basal metabolic rate?
________________________________________________________________________________________________
________________________________________________________________________________________________
________________________________________________________________________________________________
________________________________________________________________________________________________
5) Explain the connection between oxygen consumption and a persons metabolic rate.
________________________________________________________________________________________________
________________________________________________________________________________________________
________________________________________________________________________________________________
________________________________________________________________________________________________
6) What hormone is primarily responsible for the regulation of resting metabolic rate? How does it work?
______________________________________________________________________________________________
______________________________________________________________________________________________
______________________________________________________________________________________________
______________________________________________________________________________________________
7) Exercise involves the generation of action potentials by the alpha motor neurons that control the skeletal muscle motor units in your body. The alpha motor neurons are ultimately controlled by the primary motor cortex of the brain
which also has input into autonomic nervous system (ANS) control centers of the brain. Increased activity of the primary motor cortex (exercise) also stimulates the ANS. What hormone/neurotransmitter is released in response to this
activity and how does it relate to metabolic rate?
______________________________________________________________________________________________
______________________________________________________________________________________________
______________________________________________________________________________________________
Revised Fall 2014
Page 118

Date:
Name:
Section:
Mouse Basal Metabolic Rate

8) Enter your group mouse oxygen consumption data in the table below, calculate the average oxygen consumption and complete the series of calculations indicated (explained in the manual).
Cage Number:______________
Mass of mouse: ______________ grams
Mouse BMR Group Data Table

Time
(min):
Starting
Volume
(usually 0
mL)
Ending Volume (mL)

when the
bubble burst
Change in
Volume
(mL)
1-
O2 Consumption
(mL/min)
Average:
Average O2 consumption (mL/min)
______
(oC)
______
2-
Chamber Temperature
3-
Barometric pressure (mm Hg)
______
4-
Correction factor (from table or calculated)
______
5-
Corrected O2 consumption (mL/min)
______
6-
O2 volume in L/min
______
7-
Heat production (kcal/min)
______
8-
Heat production per hour (kcal/h)
______
9-
Mouse mass (g)
______
10- Surface area (m2)
______
11- Heat production (kcal/h/m2)
______
12- Mass specific BMR (kcal/h/g)
______
9) Complete the class data mouse BMR table below:
Class Data Mouse

Lab. Group
Heat Production (kcal/hr)
Mass of
Mouse (g)
Surface Area
(m2)
Heat Production
(kcal/hr/m2)
Mass Specific
BMR (kcal/hr/g)
1
2
3
4
5
6
Class
Average
Revised Fall 2014
Page 119
Name:
(continued)
10) Using your data, calculate the Surface to Mass ratios for the class average mouse:
Mouse Surface Area to Mass Ratio:
________________________________
Do the math! Surface area divided by mass.
Comparing Mouse and Human BMR Measurements

11) Rewrite the class average metabolic rate data for the human and mouse in the table below to make it easier to
compare the data:
BMR Measurement
Human
Mouse
Heat production (kcal/h)

Surface Area-Specific Heat
Production (kcal/h/m2)
Mass Specific Heat Production (kcal/h/g)
Surface area to Mass Ratio
12) Why is human heat production (kcal/h) greater than mouse heat production (kcal/h)?
______________________________________________________________________________________________
______________________________________________________________________________________________
______________________________________________________________________________________________
13) Why is human mass-specific heat production (kcal/hr/g) less than mouse mass-specific heat production
(kcal/h/g)?
______________________________________________________________________________________________
______________________________________________________________________________________________
______________________________________________________________________________________________
Revised Fall 2014
Page 120
Name:
(continued)
14) If surface area is the largest contributor explaining the difference between the human and mouse mass-specific
metabolic rates, explain your surface area-specific metabolic rate data.
______________________________________________________________________________________________
______________________________________________________________________________________________
______________________________________________________________________________________________
______________________________________________________________________________________________
Revised Fall 2014
Page 121
Revised Fall 2014
Page 122
Biology 335 Human Physiology: Renal
Laboratory #14 Renal Physiology

The Function of the Nephron
descending and ascending portions), and the distal
convoluted tubule. Two arterioles are associated
each glomerulus: an afferent arteriole feeds the gloTo understand the three nephron processes responsible merular capillary bed and an efferent arteriole drains
for urine formation:
it. These arterioles are responsible for blood flow
1. Glomerular filtration
through the glomerulus. Constricting the afferent arte2. Reabsorption
riole lowers the downstream pressure in the glomeru3. Secretion
lus, whereas constricting the efferent arteriole will
increase the pressure in the glomerulus. In addition,
the diameter of the efferent arteriole is smaller than
Background
the diameter of the afferent arteriole, restricting blood
flow out of the glomerulus. Consequently, the pressure
Waste products resulting from the metabolic proin the glomerulus forces fluid through the endothelium
cessing of nutrients are either recycled by the liver,
of the glomerulus into the lumen of the surrounding
excreted by the liver into the bile, exhaled, or filtered glomerular capsule. In essence, everything in the
out of the blood and exblood except the cells and
creted in the urine. The
proteins are filtered
filtration of the blood and
through the glomerular
production of urine is acwall. From the capsule,
complished by the kidthe filtrate moves into the
neys. Each kidney conrest of the renal tubule for
tains a million or so nephprocessing. The job of the
rons that are composed of
tubule is to concentrate
two major parts: a glomerthe urine and reabsorb all
ulus and a renal tubule.
the beneficial substances
The glomerulus is a tanfrom its lumen while algled capillary knot that
lowing the wastes to travel
filters fluid from the blood
down the tubule for elimiinto the lumen of the renal
nation from the body.
tubule. The function of
the renal tubule is to proThe nephron performs
cess that fluid, also called
three important functions
the filtrate. The beginning
to process the filtrate into
Figure 1. The Nephron and Tubule Cell Types urine: glomerular filtration,
of the renal tubule is an enlarged end called the glomertubular reabsorption, and
ular capsule (Bowmans capsule), which surrounds the tubular secretion. Glomerular filtration is a passive
glomerulus and serves to funnel the filtrate into the
process in which fluid passes from the lumen of the
rest of the renal tubule. Collectively, the glomerulus
glomerular capillary into the glomerular capsule of
and the glomerular capsule are called the renal corpus- the renal tubule. Tubular reabsorption moves most
cle.
of the filtrate back into the blood, leaving principally
salt water plus the wastes in the lumen of the tubule.
As the rest of the renal tubule extends from the gloSome of the desirable or needed solutes are actively
merular capsule, it becomes twisted and convoluted,
reabsorbed, and others move passively from the luthen dips sharply down to form a hairpin loop, and
men of the tubule into the interstitial spaces. Tubular
then coils again before entering a collecting duct.
secretion is essentially the reverse of tubular reabStarting at the glomerular capsule, the anatomical
sorption and is a process by which the kidneys can rid
parts of the renal tubule are as follows: the proximal
the blood of additional unwanted substances such as
convoluted tubule, the loop of Henle (including the
creatine and ammonia.
Objectives
Revised Fall 2014
Page 123
Biology 335 Human Physiology: Renal
The reabsorbed solutes and water that move into the

interstitial space between the nephrons need to be returned to the blood, or the kidneys will rapidly swell
like balloons. The peritubular capillaries (vasa recta) surrounding the renal tubule reclaim the reabsorbed substances and return them to general circulation. Peritubular capillaries arise from the efferent
arteriole exiting the glomerulus and empty into the
veins leaving the kidney.
Start the PhysioEx program as described in previous
laboratories. Choose the Renal System Physiology
module.
Overview
Getting Started
1.
2.
3.
4.
5.
Insert the PhysioEx 9.0 CD-ROM into the CDROM drive of the computer or access the PhysioEx folder on the desktop.
If you started with the CD-ROM a browser window with the PhysioEx opening page should
Then click on Access PhysioEx 9.0 to start the
program.
Once the PhysioEx 9.0 windows opens click on
Exercise 9: Renal System Physiology.
Beginning with the Overview, complete the Activities. At the end of each activity you are given
so, and submit to your instructor. Save the files
with unique file name such as:
Hallsec03pex-09-01
Hallsec03pex-09-02
Hallsec03pex-09-03
Etc.
Make sure the filename includes your name, section number and the exercise (-09) and activity
Revised Fall 2014
Page 124

Laboratory Manual Fall 2014

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Rhode Island College Biology 335

Revised Fall 2014

1. Safety in the Laboratory

2. Measurements and Computations

3. The Cell: Transport Mechanisms and Cell Permeability

5. Physiology of Skeletal Muscle

6. Skeletal Muscle Physiology: Computer Simulation

8. Cardiology with a Vertebrate Heart

9. Electrical Properties of the Heart

10. Circulatory Physiology

11. Mechanisms of Breathing

12. Restrictive and Obstructive Lung Disease

13. Basal Metabolic Rate

14. Renal Physiology The Function of the Nephron

Revised Fall 2014

Biology 335 Human Physiology: Safety

Revised Fall 2014

Biology 335 Human Physiology: Safety

as you would like to find it. Your colleagues and

Fire. When entering the laboratory for the

Chemical spills. Clear the

Eye Wash Station

Skin injuries. Immediately report accidents that puncture,

Long hair, long or baggy sleeves, large bracelets, and

Revised Fall 2014

First Aid Kit and Fire Blanket

Biology 335 Human Physiology: Measurements and Computations

Review the metric system.

Table 1. Metric and English Equivalents

I. The Metric System

thousand (times) [103]

Revised Fall 2014

Revised Fall 2014

To convert a Celsius temperature to Fahrenheit, use the

For instance, assume that for every liter of blood

Solve the problems found on the accompanying worksheet.

II. Computations and Presentation of Data

C. Calculation of an Arithmetic Mean

Biology 335 Human Physiology: Measurements

= number of data values in the group

Experimental thyroid mass (mg):

177.5 mg is the experimental mean.

171 mg is the control mean.

For instance, a group of six rats was given thiouracil

x = sum of all individual data of one group

By comparing the two arithmetic means, we would

Biology 335 Human Physiology: Measurements

Use the appropriate graph type

IV. Significant Figures

(2) Zeroes between nonzero digits are significant:

(4) Zeroes to the right of a decimal point in a number

(5) When a number ends in zeroes that are not to the

190 may be 2 or 3 significant figures,

Biology 335 Human Physiology: Measurements

last remaining digit is left as it is. For example,

12.4 is rounded to 12.

(4) If the digit to be dropped is 5 and is followed only

This rule means that if the digit to be dropped is 5

C. Rules for rounding off numbers

This section on significant figures was taken in part from:

Worksheet score: ____________

I. The Metric System

7. Your rats are to be fed a special diet in which each 70 g

in 10 seconds (use your left index finger applied to the radial