Topics in Biology Lab Manual-2015

Topics in Biology
Laboratory Manual
Topics in Biology Laboratory Manual
TABLE OF CONTENTS
LAB EXERCISES
INTRODUCTION TO BIOLOGY 100 LABORATORY ............................................................................ 3
BIOLOGICAL CLASSIFICATION ........................................................................................................... 16
THE THREE DOMAINS OF LIFE & CELL STRUCTURE AND FUNCTION ...................................... 23
DIFFUSION AND OSMOSIS .................................................................................................................... 34
DNA: REPLICATION, TRANSCRIPTION, & TRANSLATION ............................................................ 42
MITOSIS AND MEIOSIS………………………………………………………………………………..49
MENDELIAN GENETICS ........................................................................................................................ 58
ORGAN SYSTEMS ................................................................................................................................... 68
PLANT ANATOMY .................................................................................................................................. 83
METABOLISM .......................................................................................................................................... 96
STARCH DIGESTION EXPERIMENT………………………………………………………………..103
LAB EXPERIMENTS
OSMOSIS EXPERIMENT ....................................................................................................................... 106
GROWTH OF BACTERIA EXPERIMENT ............................................................................................ 108
ALLELOPATHY EXPERIMENT ........................................................................................................... 110
VEGETATION ANALYSIS EXPERIMENT .......................................................................................... 113
SUPPLEMENTAL MATERIAL
SCIENTIFIC WRITTEN REPORT GUIDELINES ................................................................................. 115
GRAPHING DATA .................................................................................................................................. 116
SCIENTIFIC WRITTEN REPORT GRADING POLICY ....................................................................... 117
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INTRODUCTION TO BIOLOGY 100 LABORATORY

THE NATURE OF BIOLOGICAL INQUIRY
Biologists, like all scientists, pursue a methodical search for information that reveals the secrets of the
natural world. In order to do so they use the SCIENTIFIC METHOD. The main objective of the Biology 100
laboratory is to introduce you to the scientific method. The scientific method is a sequence of steps, each depending
on the previous one.
The first step of the scientific method is making OBSERVATIONS. Observations are the facts about the
unknown. Observations can be quantitative or qualitative. Quantitative observations are measurable ones, such as
the mass, size, volume, distance, and so on. Qualitative observations are immeasurable facts such as color, texture,
and shape. For example, you observe that often people with lung cancer are also smokers.
The second step of the scientific method is asking QUESTIONS in regards to the observations that were
made. Each one of the raised questions could be the basis for a particular hypothesis. In our hypothetical example,
you may ask “Does smoking increase the risk of lung cancer?
The third step is forming a HYPOTHESIS. A hypothesis is a testable and predictable explanation of the
observed unknown process or phenomenon. The hypothesis is usually expressed with the predicted outcome. This is
called the “If-then” process. Your hypothesis may be “If someone smokes cigarettes, then she/he is more likely to
develop lung cancer compared to a non-smoker.
The forth step of the scientific method is testing the hypothesis by experiments, models, and observations.
Hypotheses must be testable in the natural world. Researchers design scientific experiments to test hypotheses in a
controlled manner. In each experiment only one variable is tested. A VARIABLE is a feature of a phenomenon (or
of an object or a process) that can change over time. Variables can be classified as independent or dependent.
INDEPENDENT VARIABLES are ones that the experimenter controls or manipulates. In our example, the
independent variable is the cigarette smoker. DEPENDENT VARIABLES are affected by the degree or presence
of the independent variable. So, development of lung cancer would be affected by smoking or not smoking. An
experiment involves tests in which conditions are carefully controlled. A CONTROL group serves as a frame of
reference. It is used to identify side effects during a test that involves an experimental group. Experiments can
contain positive controls and/or negative controls. If the control group experiences all of the same conditions as the
experimental group except for the studied variable, it is a NEGATIVE CONTROL. In the above example, cigarette
smokers form the experimental group (exposed to the cigarette smoke), whereas the non-smokers make up the
negative control group. If the control group experiences all of the same conditions as the experimental group, but
also receives treatment that is sure to give us a positive result, it is a POSITIVE CONTROL. Our example does not
have a positive control, but an example would be to have a third group of individuals be exposed to a chemical that
is known to cause lung cancer. This is obviously unethical, so we’ll stick with the negative control only.
The fifth step of the scientific method is repeating the test or the whole process. Whether the hypothesis
was supported or refuted, repeating is required. If the results of the experiment supported the hypothesis, repeating
the experiment several times more will produce more support and will increase the credibility and the usefulness of
the hypothesis. On the other hand, if the hypothesis were refuted, the researcher must check to see if something has
gone wrong. Maybe the hypothesis is not a correct one. The researcher then must start from the beginning, making
more observations, asking other questions, and coming up with a new hypothesis.
The last step of the scientific method is to analyze and report the test results as well as the conclusions.
Let’s say that in the above example, the test results showed that 20% of the smokers and 1% of the non-smokers
developed lung cancer. Since the percentages are obviously different, we can conclude that people who smoke
cigarettes are more likely to develop lung cancer compared to non-smokers.
Throughout this quarter you will be using the steps of the scientific methods to solve unknowns.
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INTRODUCTION TO BIOLOGY LABORATORY TECHNIQUES

The laboratory (and all of the equipment it contains) is to the biologist what a brush and easel are to a
painter – tools of the trade. Biology is not a spectator sport; rather, to really learn the material you must become
actively involved in doing biology. The function of the Biology 100 laboratory, therefore, is to allow you to do a
little biology. The exercises we will conduct this quarter are not complex, nor are they of Nobel Prize significance,
but they still require a bit of sophistication in terms of laboratory techniques, and today’s introductory lab exercises
will focus on these techniques.
What biologists do in the laboratory is make observations and collect appropriate data. Before you are able
to make observations or collect data, you will have to become skilled at using the basic equipment required to
measure, weigh, and observe various biological phenomena- equipment such as microscopes, balances, flasks, and
the like.
THE MICROSCOPE
The microscope will be the basic tool for many of your laboratory exercises this quarter. Microscopes are
very expensive precision instruments which require special handling, so pay particular attention to the following
guidelines concerning microscope care and use.
MICROSCOPE FUNCTION
Before learning how to use the microscope correctly, you need to know the names and functions of a few of
the basic parts. Refer to the handout for an illustration of the parts of the compound microscope described below.
The most familiar parts of the microscope are the LENSES; in the compound microscope there are two sets of
lenses. The OCULAR LENSES are the lenses to which you put your eyeballs, and the OBJECTIVE LENSES are
the lenses closer to the specimen. Note that the magnification of the ocular lenses is not adjustable (it is set at 10x),
while there are at least three (sometimes four) objective lenses, each with different magnification (4X, 10X, 40X,
and 100X). The choice of a particular objective lens is made by simply rotating the REVOLVING NOSEPIECE
until the desired lens “clicks” into position. The highest objective (100X) is the oil immersion lens, and will not be
used by you the entire quarter. To determine the total magnification of the specimen you are viewing, simply
multiply the powers of the two lenses are using (ocular x objective = total).
In order to be able to effectively view your specimen, you need to be able to adjust both the light level and
the focus of the lenses. The light can (and should) be adjusted several ways. First, there is a control knob either on
the microscope or as part of a separate light box. Second, there is an IRIS DIAPHRAGM immediately below the
stage. The focus is changed with the large knobs located along either side of the scope. The COARSE
ADJUSTMENT KNOB is the larger knob, and the smaller knob is the FINE ADJUSTMENT KNOB. As the
names imply the coarse adjustment knob moves the stage a large distance with a single turn, while the fine
adjustment knob moves the stage very little with each turn. To move slides around on the stage the two mechanical
knobs located on or under the stage will give you much more precise control over the direction and amount of
movement than using your fingers. This takes practice but is learned rather quickly. Your instructor will go over the
use of the microscope in detail.
MICROSCOPE CARE
Always follow these simple rules to make sure you do not damage the microscopes:
1. Whenever you pick up or carry a microscope, grip firmly with both hands and be especially careful to
avoid bumping or jarring it.
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2. Always set the scope down gently, both on the bench top and in the storage cabinet.
3. Do not place the scope too close to the edge of the bench, and make sure the area directly around the
scope is cleared of all books, beakers and other materials.
4. Make sure the electrical cord is out of the way where it won’t get inadvertently snagged and pulled off
the table.
5. Clean lenses with Lens Paper only! Never use paper towels or lab tissues to clean lenses, as they will
scratch the lenses. Lens paper will always be available and should be used frequently!
6. Never use lens paper for general clean up; use paper towels or lab tissues provided for cleaning other
(non-lens) parts of the microscope, wiping off slides, cleaning bench tops, etc.
7. Always use a cover slip (explained later) when viewing specimens mounted in water.
8. Before returning your microscope to your storage cabinet:
a. Remove the slide from the stage and put back in the appropriate folder
b. Neatly coil the electrical cord
c. Wipe off any liquids which may be on the scope (especially the stage and lenses)
d. Rotate the lowest power objective lens into place
e. Lower the stage to its lowest position
MICROSCOPE USE
In addition to the points raised above under MICROSCOPE CARE, you should always follow the
guidelines listed below when using the microscope. Follow these rules every time you place a new slide on the
microscope!
1. Raise the nosepiece or lower the stage using the coarse adjustment knob. This maximizes the distances
between the ocular lens and the stage, thus providing greater access to the stage.
2. Rotate the nosepiece so that the lowest power objective (4X) is in the operating position.
3. Open the iris diaphragm approximately half way.
4. Turn the light source to the lowest setting.
5. Place the slide to be viewed on the stage. With the naked eye, position the specimen directly above the
center of the condenser (the light will shine on the specimen when it is in the correct position).
6. Raise the condenser by means of the condenser knob until the top of the condenser is approximately
the thickness of a piece of paper beneath the slide. This step is very important for accurate focusing!
7. Rotate the coarse adjustment knob to bring the stage lens close to the objective lens. Keep turning the
knob until it stops (on low power the lens will not touch the slide).
8. View the specimen through the eyepiece. Slowly turn the coarse adjustment knob until the specimen
comes into focus, then use the fine adjustment knob to bring it into sharp focus. You should not have
to use the coarse adjustment knob at all after this step.
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9. The iris diaphragm may now be opened or closed to adjust the light level, thus providing more contrast
and making the viewing of your specimen easier. You should always experiment with different light
levels as you change specimens or magnifications.
10. Once the specimen is in sharp focus, it is now possible to rotate the nosepiece to other objectives
without changing the position of the coarse adjustment knob. You should only have to focus with the
fine adjustment knob. Turning the coarse adjustment with higher power objective lenses in place can
lead to broken slides and damaged scopes, so ALWAYS BEGIN FOCUSING WITH THE LOWEST
POWER OBJECTIVE, then move up to higher powers where only the fine adjustment is used.
11. Repeat this process (1-10) for each successive slide you view. Do not assume that the focus of the
optimal light levels will be similar from slide to slide.
12. ALWAYS report any sort of problem you may have been having to the instructor immediately.
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EXERCISE 1: Identify the following Microscope Parts (1 point)

Label the parts of the microscope
EXAMINING SPECIMENS WITH THE MICROSCOPE

Now is your chance to actually use the microscope. For those of you who have not used a microscope
before (and those of you who haven’t used it correctly), the microscope will introduce you to a new world of
organisms and levels of organization – the microscopic world.
EXERCISE 2: Examination of Colored Threads (1 point)

To introduce you to the microscope and its functions, we will first look at something familiar to you – silk
threads. Remember to follow the guidelines listed above for handling and focusing of the microscope.
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Materials:
Prepared slide of colored threads
Procedure:
1. Obtain a slide of colored thread.
2. Focus on the thread on scanning power, then move up to higher powers. If your microscope has a
100X objective (oil immersion; white ring) lens, do not use it.
3. Draw the thread at 40X, 100X and 400X total magnification.
Questions:
By moving the fine focus in and out, it is possible to tell which thread lies on top, in the middle, and on the bottom?
1. Which color thread is on top? ____________ Middle? ____________ Bottom? ____________
2. How do the threads compare to what you can see with your naked eyes?
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3. List all of the objective lenses on your microscope, and determine the total magnifications.
OBJECTIVE TOTAL MAGNIFICATION
___________ ___________
___________ ___________
___________ ___________
EXERCISE 3: Examination of the Letter “e” (1 point)

To get an idea of how the lenses distort the orientations of objects, we will look at another familiar object,
the letter “e” from newsprint.
Materials:
Prepared slide of the letter “e”
Procedure:
1. Obtain a slide of the letter “e”. Place it on the stage and draw the letter “e” as it appears to your naked
eye. _________
2. Focus on low power and draw letter “e” as it appears through the microscope. ___________
3. Move the slide to the right – in what direction does the image appear to move?
4. Move the slide toward you – in what direction does the image appear to move?
5. Practice focusing by increasing the magnification. At higher powers you lose track of the letter and
begin to see the wood fibers which make up the newspaper, along with some large patches of ink.
EXERCISE 4: Preparation and Examination of Wet Mounts (2 points)

During the course of Biology 100 labs, you will be asked to prepare your own slides rather than using
previously prepared slides. The type of preparation you make is called a WET MOUNT because you mount your
specimen in a liquid (usually water). The wet mount is covered with a small piece of glass called a COVER SLIP to
minimize evaporation, facilitate focusing, and to prevent the liquid from ruining the objective lenses. In the current
exercise you will practice making wet mounts of various things which interest you. Realize, though, that only small,
relatively thin specimens show up well in a microscope. Try making slides of several objects; this will give you
practice at making wet mounts, and will allow you to practice your microscope techniques. Do not hesitate to ask for
help if you encounter any problems!
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Materials:
1 Microscope slide
1 Cover slip
Dropper bottle of water
Miscellaneous specimens (hair, insects, dust, paper, etc.)
Procedure:
1. Obtain a microscope slide and a cover slip. Place a drop of water on the slide (usually in the center).
2. Find an object which you want to view. Place the specimen in the center of the drop of water
(sometimes it also helps to place a second, smaller drop on top of the specimen) Note: You may want
to start with a piece of hair plucked from your head. First make a mount of the end containing the root.
Then make a mount of the other end. Search for a hair with “split ends” and see how these compare to
“normal ends”.
3. While holding the cover slip between your thumb and forefinger at a 45 angle to the slide, place one
edge of the cover slip in the drop of water directly over the specimen (this pushes out most of the air
bubbles).
4. Draw what you see at 40X, 100X, and 400X total magnifications.
5. SLIDE CLEANUP PROCEDURE
a. REMOVE ANY SOLID ITEMS FROM THE WET MOUNT (E.G., HAIR, LEAVES) AND
DISPOSE THEM INTO THE TRASH.
b. RINSE THE COVER SLIP WITH WATER OVER THE STRAINER AND PLACE IT IN THE
GLASS WASTE CONTAINER.
c. RINSE THE MICROSCOPE SLIDE WITH WATER OVER THE STRAINER AND EITHER RE-
USE IT FOR ADDITIONAL WET MOUNTS OR PLACE IT IN THE SLIDE DRYING RACK.
6. Find two other objects to view. You may want to look at an insect such as a fly or an ant, for example.
You could also try some skin from your hand if you have a callus or blister, or some dust from the
bench tops. Use your imagination, but do not harm anyone in the process! Draw what you see at 40X,
100X, and 400X total magnifications.
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7. The goal here is to practice making wet mounts and using the microscope (changing light levels,
changing magnification, focusing, etc.).
EXERCISE 5: Introduction to Other Laboratory Materials (1 point)
In this quarter, you will make careful measurements using the International System of Measurements (SI),
commonly called the metric system. SI is used as the official system of measurement by most countries and is used
by all scientists worldwide.
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Metric Units of Measurement
Quantity Metric Unit Symbol

Mass Gram g
Length Meter m
Volume Liter l
Temperature Celsius C
Some Divisions of Metric Units
Prefix Symbol Value Prefix Symbol Value

Mega M one million Micro  one millionth
Kilo k one thousand Milli m one thousandth
Hecto h one hundred Centi c one hundredth
Deka da ten Deci d one tenth
The following exercise should acquaint you with the more common laboratory equipment and units of
measure.
Materials
Metric ruler
Graduated cylinder - 10 ml
Balance
Test tube
Forceps
Procedure:
1. Obtain a test tube and graduated cylinder. Using the graduated cylinder, determine how much water
the test tube can hold.
Volume of test tube = _______ ml
2. Now measure the dimensions of the test tube with the metric ruler.
Length = ________ mm Diameter = _______ mm
3. Now determine the weight of the test tube using the balance (your instructor will explain how this is
used).
Weight = ________ g
4. Forceps are like tweezers, and are used to handle small, fragile, hot, toxic or sterile objects. Obtain a
pair of forceps and perform similar measurements on them.
Weight = ________ g Length= _______ mm
5. EXERCISE 5 CLEANUP
a. POUR THE WATER OUT OF TEST TUBE AND PLACE THE TEST TUBE UPSIDE-DOWN IN THE TEST
TUBE DRYING RACK. THERE SHOULD BE A LAYER OF PAPER TOWELS UNDER THE RACK TO
ABSORB THE WATER DRAINING FROM THE TUBE.
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EXERCISE 6: Observations…questions…hypotheses… (4 points)

In this lab activity, you will begin practicing the steps of the scientific method. You will be making
observations, asking questions, forming hypotheses, and conducting simple experiments to test these hypotheses.
You are presented with a set of sealed boxes, each with an object inside. Your task is to conduct a scientific study to
identify the objects. There are balances, stethoscopes, empty sealed boxes, etc., to help with your observations.
Work in groups of two or three and record the number of the box you have chosen on the box observation
sheet (pages). Record as many (5-7) quantitative characteristics (length, width, weight, etc.) and qualitative
characteristics (texture, shape, material, etc.) of the object inside the box as possible . These are your observations,
so be thorough!
Summarize all the characteristics you are able to infer about the object and make a hypothesis about the
identity of the object in the box. YOU ARE NOT GRADED BY GETTING THE "RIGHT" ANSWER, BUT BY
THE CLEVERNESS AND VARIETY OF YOUR OBSERVATIONS. There are many boxes to test but do only one
(not including the “E” boxes).
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BOX OBSERVATION SHEET #1 (2 points) Name

Box number
Remember that you are interested in the object, not the box!
Technique Observations Conclusion
What did you The actual What does this

do to make your results from tell you about
observations? what you did. the object?
Total of your conclusions: What do you think the object is?____________________
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Questions (1 point each):
1. How confident are you in your conclusions? Use your findings (give at least 2 of your observations in
the box observation sheet) to explain your answer.
2. Assuming that no one knows what is in the box and you can’t open the box, what could you do to
increase your confidence in your conclusions?
Turn in pages 3-15 for this lab.
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BIOLOGICAL CLASSIFICATION
ANALYTICAL KEYS
Human beings are always naming and organizing the things around them. This process of classification
helps individuals communicate with each other and helps them to understand their world better by creating patterns
and relationships.
TAXONOMY is the field of biological classification. The traditional taxonomy is based on the
BINOMIAL CLASSIFICATION SYSTEM that was developed by the Swedish naturalist Carl von Linne in the
early 1700’s. His system has been reasonably successful in naming millions of organisms that have been classified.
Today molecular biology has extended the field of taxonomy. DNA, RNA, and protein profiling are used to
reveal more information about life on this planet.
With millions of different organisms identified so far, it is impossible for anyone to know them all.
ANALYTICAL KEYS are tools that enable someone to identify an unknown organism. An analytical key (is based
on traditional taxonomy) consists of a series of questions that ask about observed characteristics of an unknown
organism. These characteristics must be visible, constant (non-changeable), and quantifiable (if the characteristic is
quantitative). As you answer each question, the key directs you to another question and then another until you reach
the final identification of the organism.
Below is an example of a simple analytical key.
Example key: Population sample – red apple, orange, lemon, grape, banana
1. a. object is spherical ......................................................... 2

b. object is not spherical .......................................... banana
2. a. object is less than 4 cm in diameter ........................grape

b. object is over 4 cm in diameter .................................... 3
3. a. object is orange in color .......................................orange

b. object is not orange in color ......................................... 4
4. a. object is yellow in color ....................................... lemon

b. object is not yellow in color ................................... apple
Notes:
The above key has four dichotomous questions and identifies five fruits.
The organization of the key is concerned with the proper question format that makes it easy to follow. This
format involves a series of questions with only two possible choices at each step. This format produces a
DICHOTOMOUS KEY.
Any characteristic can be used.
The exercises in this lab are based on the traditional taxonomy.
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EXERCISE 1: Make a Dichotomous Key (Part I) and

Anwer the Questions on Page 18 (3 points)
Name ____________________________
Key # 1 (2 points)
In the space below, construct an analytical key for the people in your lab group.
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Questions (0.5 points each):
1. Would you be able to identify someone else (not from your group) with the key that you have made in
exercise # 1? What aspects of your key could be a problem?
2. Can you make any suggestions to modify the statement “any characteristic can be used”?
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EXERCISE 2: Make a Dichotomous Key (Part II) (2 points)
Name ____________________________
Key # 2 (2 points)
In the space below, construct an analytical key for the bird species you have been assigned.
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EXERCISE 3: Make a Dichotomous Key (Part III) and

Anwer the Questions on Page 21 (3 points)
Name ____________________________
Key # 3 (2 points)
Materials
Set of plastic bugs
Procedure:
1. Organize the plastic bugs into ten groups or piles.
2. Construct an analytical key for the groups of objects you have been assigned. Each group is to be
treated as one genus in your key. Be sure to indicate the group names in your key. (When constructing
your key, you can not use the names of the objects in your statements. Use observable physical
descriptions).
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When you were organizing and constructing your analytical keys you were actually using the scientific method.
Remember that the scientific method involves observation, question, hypothesis, testing, repeating, and reporting.
1. What were your observations (list 2 or more)?
2. What was your question (list 2 or more)?
3. What was your hypothesis?
4. Did you test your hypothesis? How?
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EXERCISE 4: Using a Dichotomous Key (2 points)
Name______________________________
Materials
“Unknown” bug from instructor
Procedure
1. After your group has completed Exercise 3, your instructor will give you another object. Run this new
object through your groups’ key from Exercise 3.
1. Does this object key out properly? Would you still end up with 10 groups?
2. What does this say about your hypothesis?
3. Assume that you were given an object that did not key out properly and you were asked to identify that
object, what should you do to your existing key?
4. Using your answer to question 3 above, what does this tell you about a scientific hypothesis and the
practice of science in general?
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THE THREE DOMAINS OF LIFE &

CELL STRUCTURE AND FUNCTION
DOMAINS OF LIFE
Although Biology 100 focuses on humans, you should have an appreciation for the diversity of organisms
on earth. There are between 2 million and 50 million species existing today, but they are going extinct faster than we
can capture and identify them!
The classification of organisms (carried out in a branch of biology called TAXONOMY) is based on
similarities between organisms. Early taxonomists recognized only two groups of living organisms, PLANTS and
ANIMALS. However, with the availability of more and more information (including molecular information from
DNA and RNA analysis) about the structure and function of organisms, taxonomists have recently realized that
more than two major groups exist. We now use a classification scheme which recognizes 3 major groups, or
DOMAINS, of organisms. These are DOMAIN BACTERIA, DOMAIN ARCHAEA, and DOMAIN
EUKARYA. The organisms in the three domains are different based on the presence or absence of organelles, types
of organelles within their cell(s), the types (or lack) of walls around cell(s), the number of cells in a mature
organism, the mode of feeding of the organism, modes of reproduction, their size and so on. The lecture portion of
the course will deal with some of the differences between the three domains with respect to the characteristics just
listed. In this lab exercise, you should familiarize yourself with the appearance and diversity of members of these
groups, as some of them may be totally foreign to you.
CELL STRUCTURE AND FUNCTION

Cells are the basic units of life. Each eukaryotic cell has substructures within them called ORGANELLES
(literally “little organs”) which carry out the processes of life. Prokaryotic cells (those within the Domains Bacteria
and Archaea) lack these internal organelles, but are still able to carry out some of the simpler processes necessary to
sustain life. For Biology 100 labs, we will consider prokaryotic cells, but will focus on eukaryotic cells.
EUKARYOTES
Domain Eukarya is divided into 4 kingdoms: PROTISTA, FUNGI, PLANTAE, and ANIMALIA.
Eukaryotes consist of larger cells (larger than prokaryotes) that are more complex than prokaryotic cells. They
(eukaryotic cells) contain an array of organelles, including the nucleus which contains the cell’s DNA. Organelles
within eukaryotic cells allow for division of duties; this “assembly line” approach to carrying out a diverse array of
cellular functions allows for greater efficiency within each cell. The organelles within each cell can be thought of as
a series of separate compartments, each with their own specialized function; cells are able to move materials into
and out of these compartments to suit their needs.
EXERCISE 1: Kingdom Protista (2 points)

The protistans are another diverse group of organisms, with over 35,000 known species that you may not be
very familiar with. The kingdom includes three major groups: the ANIMAL-LIKE PROTOZOANS, the PLANT-
LIKE PROSTISTS, and the FUNGUS-LIKE PROTISTS. They all share the characteristic of being free-living,
single-celled eukaryotes (i.e., they all have internal organelles). Kingdom Protista includes several important
parasites. As an example of the size and simplicity of protistians, you will look at Euglena, a plant-like protistan,
and Paramecium, an animal-like protistan.
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Materials:
Prepared slide of Euglena
Wet mount of living Euglena (on display)
Wet mount of living Paramecium (on display)
Prepared slide of Paramecium
Procedure:
1. View the Euglena set up on the demo scope under different magnifications, noting how they move
through the water with great ease. If they are moving too fast to see clearly, you may want to ask the
instructor to add methylcellulose (a viscous liquid that slows their movement) to the viewing medium.
a. Obtain a prepared slide of Euglena.
b. Observe and draw the Euglena.
2. Repeat the above procedure (live organism on demo scope and prepared slide) for Paramecium.
3. a. What structures in Euglena are found in plant cells?

b. What structures in Euglena are found in animal cells?
4. a. What structures in Paramecium are found in plant cells?

b. What structures in Paramecium are found in animal cells?
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EXERCISE 2: Kingdom Fungi (1 point)

There are over 100,000 species of fungi. The kingdom Fungi is characterized by eukaryotic, multicellular,
filamentous (thread-like) or unicellular (yeast-like) individuals which gain their energy in a unique way: they secrete
enzymes outside of the body and breakdown organic matter, and then absorb the “digested food” into their body for
nourishment. They usually function as decomposers of dead material, but some act as parasites on living material.
Fungi include the common mushrooms as well as molds, mildew and yeasts.
Materials:
Specimens of preserved fungi
Prepared slide of Penicillium
Procedure:
1. Examine the preserved specimens at the side table. Be able to identify them to kingdom.
a. Obtain a prepared slide of Penicillium.
b. Observe and draw Penicillium.
EXERCISE 3: Kingdom Plantae (2 points)

Plants are characterized as being eukaryotic, multicellular, and able to produce their own food through the
process of photosynthesis. You are already familiar with numerous types of plants, so this exercise will focus on
plant cells and the organelles found within them. Pay particular attention to the organelles shared by both plants and
animals and to those absent from animal cells.
Materials:
Preserved specimens of plants
Living Elodea leaves
Onion
Razor blades
Microscope slides
Coverslips
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Methylene blue dropper bottle
Procedure:
1. Examine the preserved specimens of plants available at the side bench. Be able to identify them to the
kingdom.
2. Remove a young Elodea leaf from the sprig. Make a wet mount with the top of the leaf facing
upwards. Examine the leaf with your microscope.
a. Note the individual cells surrounded by the visible cell wall. The plasma membrane is
located just inside the cell wall.
b. Observe the numerous chloroplasts within each cell. They appear as moderately-sized green
spheres. You may notice the chloroplasts begin to move. This movement is called cytoplasmic
streaming. Chloroplasts move in order to optimally absorb light (in visible light range) which
is important in photosynthesis. What is the function of the chloroplast? Where in the cell are
the chloroplasts located: toward the perimeter or centrally?
c. The chloroplasts may be pushed to the periphery of the cell by the central vacuole. This large
sac serves to store water, ions, enzymes and waste products. Is a central vacuole obvious in
any of your cells?
d. Use the fine focus adjustment knob of your microscope and search for a nucleus. You may
add a drop of methylene blue to the slide to enhance its visibility.
e. Draw an Elodea cell using the high power objective lens and label the organelles you
observed: cell wall, chloroplasts, the location of the central vacuole and the nucleus (if you
were able to observe it).
26
a. REMOVE ANY SOLID ITEMS FROM THE WET MOUNT (E.G., HAIR, LEAVES) AND
DISPOSE THEM INTO THE TRASH.
b. RINSE THE COVER SLIP WITH WATER OVER THE STRAINER AND PLACE IT IN THE
c. RINSE THE MICROSCOPE SLIDE WITH WATER OVER THE STRAINER AND EITHER RE-
4. Make a wet mount using a piece of onion epidermis. Examine the tissue.
a. Place a small drop of methylene blue at the edge of the coverslip. Hold the edge of a paper
towel at the opposite edge of the coverslip. This will draw the fluid toward the paper towel,
staining the onion cells.
b. The nucleus should appear as a darkly stained object in the center of the cell. Draw a stained
onion cells using the high power objective lens and label the organelles you observed:
nucleus, cell wall, cell membrane, the location of the central vacuole.
c. Did you see any chloroplasts in the onion cells? Explain.
5. Repeat the slide cleanup procedure.
EXERCISE 4: Kingdom Animalia (2 points)

Animals are characterized as being eukaryotic, multicelluar, and requiring food from an outside source
(heterotrophic). Again, you are already familiar with types of animals, so we will focus on animal cells here.
Materials:
Toothpicks
Microscope slides
Coverslips
Methylene blue
Prepared slides of human cells
Preserved animal specimens
27
Procedure:
1. Examine the preserved specimens of animals available at the side bench. Be able to identify to
kingdom.
2. Make a wet mount of epithelial cells from the inside of your cheek. Your instructor will demonstrate
this for you.
a. As with the onion cells, place a drop of methylene blue at the edge of the coverslip and draw
it through the sample with a paper towel.
b. Locate the stained nucleus.
c. Draw the cheek cell and label the organelles you observed: nucleus, cell membrane, the
location of the cytoplasm.
d. How does the size and shape of the human epithelial cell compare with that of the onion
epithelial cell?
e. Why do the plant cells have a more consistent shape than the human cells?
f. To gain an appreciation of the diversity among animal cells, view several of the available
prepared slides.
g. Draw two type of animal cells on the following page, noting any organelles that may be
visible.
28
3. SLIDE CLEANUP PROCEDURE FOR CHEEK CELLS
a. USING GLOVES, SEPARATE THE MICROSCOPE SLIDE AND COVER SLIP. RINSE BOTH
THE SLIDE AND COVER SLIP OVER THE STRAINER WITH THE 10% BLEACH
SOLUTION PROVIDED IN SQUEEZE BOTTLES AT THE SINK.
b. RINSE THE COVER SLIP OVER THE STRAINER WITH WATER AND PLACE IT IN THE
c. RINSE THE MICROSCOPE SLIDE OVER THE STRAINER WITH WATER AND EITHER RE-
PROKARYOTES
Prokaryotes are divided into two domains: Archaea and Bacteria. Prokaryotes are quite diverse, but they
have some distinct characteristics in common: they all lack internal organelles. They are all UNICELLULAR
(exist as single-celled individuals), but many species grow in very large aggregations, or colonies. Some are able to
carry out photosynthesis while others must consume tissues of other organism as a food source.
Domain Archaea is divided into 3 kingdoms. You will get information about these 3 kingdoms in lecture
(Ch. 21). The organisms in domain Archaea are mostly anaerobes, living in extreme habitats such as high heat or
areas with high salt concentration. Since early earth had such extreme conditions, Archaea is thought to contain
organisms similar to the earliest living cells on this planet.
Domain Bacteria is divided into 12 kingdoms. Chapter 21 of your text book gives information about some
of the 12 bacterial kingdoms. Bacteria are very small - about 1000 times smaller than a eukaryotic cell (typical cell
in your body). They are so small that a magnification of 1000X is barely sufficient to observe them. Despite their
size, they are still very important to both plants and animals. Bacteria are both good and bad, but we usually only
notice when they are causing problems such as diseases, spoiled food, soured milk, and the like. However, they play
very important roles in the decay process of dead organisms (plant and animal) and in cycling nutrients through
ecosystems. We even house billions of them in our digestive tract to help us digest food! Since bacteria are very
small, they require a special technique to view them under the light microscope. Hence, we will set up several
display scopes for you to view. The bacteria on display are of three general types: BACILLI are rod-shaped, COCCI
are spherical, and SPRILLI are helical (S&T Figures 21.3, 21.4). See Chapter 21 for some examples of the types and
roles of various prokaryotes around you.
29
EXERCISE 5: Domain Bacteria (1 point)

Materials:
Prepared slide of bacteria (on display)
Yogurt
Procedure:
1. Observe the slides of bacteria on the demo microscopes.
a. Identify and draw the three types of bacteria.
Bacilli Cocci Spirilli
2. Bacteria play a vital role in many aspects of our lives, including food production. View the yogurt wet
mount on the demo microscope. What types of bacteria can you identify? Identify and draw.
30
EXERCISE 6: Bacteria in our environment (2 points)
Materials:
Petri dish containing agar (substance that support the growth of bacteria and fungi).
Sterile cotton swabs
Sharpies
Procedure (work individually):
Week 1
1. Decide which three parts of your local environment you wish to sample for bacteria.
2. Obtain a sterile agar plate. Keep it sterile, Do not open yet!
3. Label the bottom of the plate with the following:

 students initials or names
 type of exposure
 date of exposure
 lab.section time (e.g., Tue 12-3)
4. Using a sharpie, divide the bottom of the plate into four quadrants. Number each quadrant.
5. Leave quadrant # 1 unexposed. Quadrant # 1 will be used as your control.
6. Expose quadrants 1-3 each to a different sample (such as your cell phone, backpack, table tops,
bathroom facilities, whatever you want to examine for the presence of bacteria). Use a separate
applicator for each sample, and throw the applicators in the trash when you are done. Do not open the
plate longer than is necessary for your exposure.
6. Cover and turn your dish upside down. Place on the area designated for your class.
7. Let sit at room temperature until the next lab period.
Week 2
1. Obtain your plate from the previous week and examine them for the density of the colonies and any
diversity you can see. Diversity will be determined by differences in colony color, shape, size and
texture. Record the diversity (how many different types of bacteria grew in each of the quadrants).
Also record the density of the colonies (how many colonies of each type of bacteria grew in each
quadrant).
a. PLACE ALL USED PLATES IN THE RED BAG IN THE BIO-HAZARD DISPOSAL CAN. DO NOT
THROW IN THE TRASH CAN!
31
Growth of bacteria Results
Type of exposure Appearance of Plate

______________________________________________________________________________________
______________________________________________________________________________________
______________________________________________________________________________________
______________________________________________________________________________________
______________________________________________________________________________________
Write a paragraph comparing the results of the different exposures. Which exposure was the most contaminated with
bacteria? Why? Use the quantitative as well as the qualitative results to support your opinion. Use the space bellow.
32
Prokaryotes Eukaryotes
Bacteria Archaea Protists Plants Fungi Animals
Cell Surface
In some
Cell Wall + + species + +
Plasma membrane + + + + + +
In many In many
Flagella species species + + + +
Cilia + + + +
Links between cells

Pili + +
Junctions + + + +
Genetic Machinery
DNA + + + + + +
Single circular chromosome + +
Multiple linear chromosomes + + + +
Organelles
Nuclear envelope + + + +
Cytoplasm + + + + + +
Ribosomes + + + + + +
Cytoskeleton + + + + + +
Endoplasmic reticulum + + + +
Golgi apparatus + + + +
Lysosomes + + +
Mitochondria + + + +
Chloroplasts + +
Central vacuole + +
*Note that a "+" indicates that it may occur in that kingdom
33
DIFFUSION AND OSMOSIS

Diffusion and osmosis are important physical properties in all biological processes. Both diffusion and
osmosis are required in part to control the movement of ions (charged atoms or molecules), water, and organic
molecules in living systems. Thus, the proper functioning of nerves, muscles, kidneys, lungs and every organ in your
body depends on diffusion and osmosis.
The driving force of diffusion and osmosis is the random movement of particles provided by kinetic
energy. KINETIC ENERGY is the energy of motion, and occurs in all states of matter (solids, liquids, and gases).
The kinetic energy increases as the temperature of the matter increases, and decreases as the temperature decreases;
hence the movement of atoms and molecules changes as the temperature changes. At a very low temperature, called
“absolute zero” (-273°C or -460°F), the kinetic energy equals zero and all motion ceases. Above absolute zero,
particles “bounce” around at random, moving in a straight path until they collide with another particle (which they
“bounce” off and proceed to travel in another direction until they bounce off another, and another, etc.).
Since biological systems always function far above absolute zero, it is safe to assume that there is always
quite a bit of kinetic energy (hence random molecular motion) within living systems. What this means, in a practical
sense, is that molecules are in a constant state of motion, even in material which (to the naked eye) looks like it is
still. A familiar example of this motion, and the effects of temperature on the amount of motion, is the rate at which
sugar dissolves in cold versus hot tea. When the tea is cold, sugar will sit on the bottom of a glass and slowly
dissolve; however, as the temperature of the tea (hence its kinetic energy) begins to increase, the rate at which the
sugar dissolves increases dramatically. This is due in part to the different rates at which the sugar molecules are
“moving away from the pile of sugar”. They still move away in cold tea, but they move much faster in hot tea due to
increased level of random motion. This molecular movement is also affected by the size of the molecules (in
general, smaller molecules will move faster than larger aggregates of molecules) and the nature of the medium they
are moving through (faster in gases than in liquids, and so on).
One of the first persons to observe the random motion of particles was Robert Brown (1827), so we now
call this motion BROWNIAN MOTION. To observe Brownian motion (and the effects of size and temperature),
conduct the following simple exercise.
EXERCISE 1: Observation of Brownian Motion (2 points)

You can observe the random motion of particles in milk (which is full of relatively large fat droplets) and in
India ink (which contains numerous, very small carbon particles).
Materials:
Microscope slide
Cover slips
Milk (in dropper bottle)
India ink solution (in dropper bottle)
Procedure:
1. Place one drop of milk on one end of a microscope slide.
2. Place one drop of the ink solution on the other end of the same microscope slide (do not let the two
solutions touch!)
3. Place cover slips over the two solutions.
4. Observe the solutions through the microscope and draw them below.
34
Milk Ink
a. RINSE THE COVER SLIP WITH WATER OVER THE STRAINER AND PLACE IT IN THE
b. RINSE THE MICROSCOPE SLIDE WITH WATER OVER THE STRAINER AND EITHER RE-
Questions:
1. Which of the suspended materials moves fastest, fat droplets or the carbon particles?
2. Why do they move at different rates?
3. Leave the slide over the light for a minute and let the temperature increase. How does this affect the
rate of movement in the ink and milk?
35
DIFFUSION
DIFFUSION is the net of movement of particles from an area of high concentration to an area of low
concentration. This movement occurs via Brownian motion: the particles in the area of high concentration tend to
move out to areas with a lower concentration. Although some also move back toward the area of higher
concentration, the majority (due to the random nature of the direction of movement) tend to move to areas of lower
concentration. Eventually, the particles would be equally distributed within the medium in they are diffused; this is
termed the “equilibrium” state. Since diffusion is driven by Brownian motion, and Brownian motion increases with
temperature, the rate of diffusion is also strongly affected by temperature. Remember the example of the sugar
diffusing in hot and cold tea? The diffusion of sugar molecules in warm tea occurs much more quickly than
diffusion of sugar molecules in cold tea simply because the rates of Brownian motion are much greater in the warm
tea. This means that sugar molecules are “escaping” from the pile of sugar and moving throughout the tea at random
at a much higher rate in warm than in cold tea.
As an analogy, imagine 15 pool balls clustered in the center of a pool table. If you were to hit the cue ball
into the group, the balls would move out of the “area of high concentration” to “areas of lower concentration,”
bouncing off of the walls and the other balls during the process. The balls move in a straight line until they collide
with something - either the wall or another ball. Although some might, by chance, return to the center of the table,
most would wind up elsewhere, “diffused across the table” in a sense. If you can imagine the balls bouncing around
and never stopping, eventually becoming equally distributed (yet still moving) across the table, you would have an
idea of the result of diffusion. In this analogy, the speed of the balls would be an indication of the temperature of the
system; the higher the temperature, the faster the balls travel, the more often they collide with each other and the
walls, the faster they spread out and reach equilibrium, etc.
As realistic example of diffusion, think about the effect of opening a bottle which contains a foul smelling
liquid or gas (vinegar, rotten food, etc) in a closed room. The gas is in high concentration in the jar, and in low
concentration in the air within the room. However, soon the “smell” (gas particles) begins to diffuse away from the
jar and out into the room. At first the smell is strongest near the bottle and diminishes as you move away from it, but
if you leave the bottle open long enough the entire room will smell the same (equilibrium has been reached). This is
diffusion.
Diffusion is not restricted to any one medium; it can (and does) occur in solids, liquids and gases. The rate
of movement is greatest in gases and slowest in solids, but it still occurs in all three. Also, any particle can diffuse,
however, the rate is affected by factors such as size (with large particles diffusing more slowly than small ones) and
chemical properties (charged versus uncharged, hydrophobic versus hydrophilic, etc.) of both the particles and the
medium.
The following exercise is designed to give you feel for rate of diffusion and the differences in rates of
movements between different molecules.
EXERCISE 2: Diffusion in a Semi-Solid (2 points)
In this exercise you will examine the movement of two dye salts diffusing within a semi-solid medium called agar,
which is very similar to the gelatin that you eat. You will compare the different rates of movement of two molecules
of different sizes: potassium permanganate and methylene blue. Potassium permanganate (KMnO4) is an ionic
compound consisting of potassium (K+) and manganate (MnO4-) ions, and has a molar mass of 158.03 g/mol.
Methylene blue (C16H18N3SCl) is a complex chemical compound with a molar mass of 319.85 g/mol. Thus, one
molecule of potassium permanganate has about half the molar mass of one molecule of methylene blue.
Materials:
Forceps
Paper disks
Petri dish containing agar
1% solution of potassium permanganate (TOXIC – wear gloves and safety goggles.)
36
1% solution of methylene blue (TOXIC – wear gloves and safety goggles.)
Procedure:
1. Obtain a Petri dish containing agar.
2. Using the appropriately labeled forceps, pick up a paper disk and dip it into the solution of
potassium permanganate, and then place it on the surface of the agar about 1cm from the edge of
the plate. Be careful not to drip any of the solution onto the agar.
3. Repeat step 2 for the methylene blue. Remember to use the appropriately labeled forceps and place
the paper disk at the opposite edge of the plate.
4. Set the plate aside and observe occasionally.
5. After 90 minutes, measure (in millimeters) the distance each of the compounds diffused from the edge of
its paper disk. Draw to scale.
a. THROW LIDS IN THE TRASH CAN.

b. PUT PLATES IN THE WASTE JAR LABELED “METHYLENE BLUE AND POTASSIUM
PERMANGANATE WASTE.”
Questions:
1. Which molecule diffused the fastest? _____________________ The slowest? ___________________

Why do you think this might be so?
2. What effect might we see if we put one plate at room temperature, another on ice, and a third on a hot
plate?
37
EXERCISE 3: Effects of Temperature and Medium on Rate of Diffusion

(2 points)
The temperature and nature of the medium has a large effect on rates of diffusion. In the current exercise,
you will examine the effects of both temperature and hydrophobic or hydrophilic interactions on the rates of
diffusion of sugar into two different solvents. Observe the following demonstration and answer the questions.
Materials:
3 sugar cubes
3 250-ml beakers
Room temperature water
Cold water
Hot water
Beaker with vegetable oil (demo)
Procedure:
1. Place 100 ml of each solution into a different beaker. Mark the beakers with label tape and a marker so
you can identify them. Number the beakers (1-3) according to the table below.
2. Place the three beakers side-by-side on the bench in front of you.
3. Drop the sugar cubes into all beakers simultaneously and begin timing how long it takes for the sugar
to completely dissolve. Fill in the table below.
a. POUR SUGAR WATER DOWN THE DRAIN AND RINSE EACH BEAKER. MAKE SURE THERE IS NO
VISIBLE SUGAR REMAINING IN THE BEAKER.
b. PLACE THE BEAKER UPSIDE DOWN TO DRAIN.
Beaker Elapsed Time Solvent Temperature

1 water cold
2 water room
3 water hot
Demo oil room
Questions:
1. In which solution did the sugar dissolve most rapidly? __________ Most slowly? ____________
Why?
2. Is sugar hydrophobic or hydrophilic? How do you know?
3. What effect would stirring the solution have on the results?
4. Give examples of molecules that diffuse into or out of your cells.
38
OSMOSIS
OSMOSIS is a special case of diffusion: it is the movement of water from an area of high water
concentration to an area of lower water concentration (in other words, osmosis is the diffusion of water). An
important point about osmosis is that it occurs across (or through) a SEMI-PERMEABLE or SELECTIVELY-
PERMEABLE MEMBRANE. A semi-permeable membrane, as the name implies, is a membrane which allows
certain molecules to pass through while excluding others (in much the same way that a sieve has pores which allows
water to pass through but prevents pasta from escaping). The plasma membrane surrounding cells is a semi-
permeable membrane, as is the dialysis tubing we are using in today’s lab: both are permeable to water, yet
impermeable to sugar (and many other molecules).
If the concentration of water is greater inside the cell than outside the cell (note that this is the same as
saying that there is a lower concentration of solutes inside the cell than outside the cell), water will move out of the
cell. In this situation the surrounding fluid is said to be HYPERTONIC to the fluid within the cell. If the
concentration of water is lower inside the cell than outside the cell (i.e., more solutes inside than outside), water will
move into the cell. In this situation the surrounding fluid is HYPOTONIC to the fluid within cell. If the
concentration of water is the same inside and outside of the cell, there will be no net movement of water into or out
of the cell, and the surrounding fluid is ISOTONIC to the fluid within the cell.
EXERCISE 4: Osmosis Across Living Membranes: Animal Cells (2 points)
In this experiment you will be exposing mammalian red blood cells (RBCs) to different concentrations of
table salt (sodium chloride, NaCl) in distilled water. They will react in much the same manner as the artificial cells
constructed above, however, the process of osmosis will occur much more quickly, the RBC cell membrane is so
fragile that if it expands too much it will burst, and they are too small to view without the aid of a microscope. There
is one test which can be conducted with the naked eye that can detect the bursting of red blood cells. Whole RBCs
make a solution so turbid that you can not easily see through the solution whereas a solution of ruptured RBCs
impart a red color but no turbidity, still allowing you to see through the solution. Thus, by holding a test tube or slide
containing a solution of RBCs against a printed page you can quickly tell if the cells have ruptured (i.e., are in a
hypotonic solution).
Materials:
3 small glass test tubes
3 microscope slides
3 cover slips
Ruler
Blood (in dropper bottle)
Distilled water
0.85% NaCl
5.0% NaCl
Procedure:
1. Thoroughly wash and rinse all vials before and after the exercise.
2. Label the pipettes, slides and test tubes for each of the solutions (water, 0.85%, 5.0%).
3. Place about 1 ml (~1 cm) of each solution into the appropriate test tube.
4. Place 2 drops of blood into each of the three tubes. Gently swirl to mix.
5. Hold each tube against this page and see if you can read through the mixture. Record your results in
the table on the following page.
39
6. Place a drop of each of the blood mixtures onto the appropriate microscope slide (use a different
pipette for each mixture), cover with a cover slip, and view in the microscope. Draw the results in the
table on the following page. (When viewing and drawing RBCs, increase the magnification to 400X
and minimize the light intensity for the best image).
a. USING GLOVES, SEPARATE THE MICROSCOPE SLIDE AND COVER SLIP.

b. RINSE THE SLIDES, COVER SLIPS, AND TEST TUBES OVER THE STRAINER WITH THE 10%
BLEACH SOLUTION PROVIDED IN SQUEEZE BOTTLES AT THE SINK.
c. RINSE THE COVER SLIPS OVER THE STRAINER WITH WATER AND PLACE THEM IN THE GLASS
WASTE CONTAINER. NO BLOOD SHOULD BE VISIBLE ON THE COVER SLIPS.
d. RINSE THE MICROSCOPE SLIDES OVER THE STRAINER WITH WATER AND EITHER RE-USE
THEM FOR ADDITIONAL WET MOUNTS OR PLACE THEM IN THE SLIDE DRYING RACK. NO
BLOOD SHOULD BE VISIBLE ON THE SLIDES.
e. RINSE THE TEST TUBES WITH WATER AND PLACE THEM UPSIDE DOWN IN THE TEST TUBE
DRYING RACK. NO BLOOD SHOULD BE VISIBLE IN THE TEST TUBES.
Solution Clear or Opaque Appearance of Cells
Water
0.85%
5.0%
Questions:
1. Which solution(s) is hypotonic to blood cells? ________________ Hypertonic? ______________

Isotonic? _____________
2. Based on these results, what do you think the salt level of your blood is?
EXERCISE 5: Osmosis Across Living Membranes: Plant Cells (2 points)

Like the animal cells observed in the previous exercise, osmosis also occurs in plant cells. However, the
behavior of plant cells in response to osmosis is remarkably different due to the cell wall. When placed in a
hypertonic solution, water moves out of the plant cell, causing the cytoplasm to shrink as the cell membrane pulls
away from the cell wall. This event is called plasmolysis. When a plant cell is placed in a hypotonic solution, water
moves into the plant cell. Unlike the animal cell, the plant cell does not burst. Rather it becomes rigid, or turgid. You
may notice your house plants beginning to wilt when you forget to water them. Once you water them, they become
turgid again and stand tall. In this exercise you will examine the effects of salt on Elodea.
Materials:
40
Microscope slide
Cover slip
Elodea leaf
Distilled water
10% NaCl solution
Pond water
Procedure:
1. Prepare a wet mount of an Elodea leaf using pond water. Draw a few cells below.
2. Prepare a wet mount of an Elodea leaf using distilled water. Draw a few cells below.
3. Prepare a wet mount of an Elodea leaf using 10% NaCl solution. Draw a few cells below.
a. REMOVE ANY SOLID ITEMS FROM THE WET MOUNT (E.G., HAIR, LEAVES) AND DISPOSE
THEM INTO THE TRASH.
b. RINSE THE COVER SLIP WITH WATER OVER THE STRAINER AND PLACE IT IN THE GLASS
WASTE CONTAINER.
c. RINSE THE MICROSCOPE SLIDE WITH WATER OVER THE STRAINER AND EITHER RE-USE IT
FOR ADDITIONAL WET MOUNTS OR PLACE IT IN THE SLIDE DRYING RACK.
Pond Water Distilled Water 10% NaCl
Questions:
1. Which solution is hypertonic to the plant cells? ______________ Hypotonic? ______________

Isotonic? _______________
2. How does osmosis in plant cells compare with osmosis in animal cells?
41
DNA: REPLICATION, TRANSCRIPTION,

& TRANSLATION
DEOXYRIBONUCLEIC ACID, or DNA, is often referred to as the blueprint for life. It contains
all the information necessary for a cell or organism to develop, function, and reproduce. This information is stored in
sequences of DNA called genes, which are in turn organized within chromosomes.
DNA is a type of NUCLEIC ACID, which is one of the four main types of biological molecules (the other
three being carbohydrates, proteins, and lipids). Nucleic acids consist of a chain of nucleotides, with each nucleotide
composed of a phosphate group, a 5-carbon sugar, and a nitrogenous base. DNA contains four different nitrogenous
bases: adenine (A), thymine (T), guanine (G), and cytosine (C). The other type of nucleic acid, RIBONUCLEIC
ACID or RNA, also contains four nitrogenous bases but uses uracil (U) instead of thymine. A DNA molecule
consists of two twisted chains of nucleotides that are connected by hydrogen bonds between the nitrogenous bases
on either strand. It is convenient to think of a DNA molecule like a ladder. If you split the ladder lengthwise down
the middle, you are left with two strands of DNA, each composed of a sugar-phosphate backbone (the side of the
ladder) with a series of nitrogenous bases (the split rungs of the ladder). A base on one strand of DNA always pairs
with its complementary base on the other strand: adenine always pairs with thymine (and vice versa) and guanine
always pairs with cytosine (and vice versa). These complementary bases (and thus the two strands of DNA) are held
together by hydrogen bonds, and the backbones twist to form a structure called a double helix. This structure eluded
scientists for decades until it was reported by James Watson and Francis Crick in 1953.
DNA is arranged differently in prokaryotes and eukaryotes. In prokaryotes DNA exists as a single circular
chromosome within the cytoplasm of the cell. Eukaryotic DNA is more complex, with multiple linear chromosomes
arranged within the nucleus of the cell. Each species has a characteristic number of chromosomes. For example, fruit
flies have 8 chromosomes, onions have 16 chromosomes, and humans have 46 chromosomes in each of their
somatic cells. The structure and function of DNA in prokaryotes and eukaryotes, however, is the same. For all
organisms DNA is the genetic information that is passed from parent to offspring, from one generation to the next.
Over the past 50 years DNA has become an important research subject for scientists and has become
familiar to the general public as well. Evolutionary biologists use DNA to trace the history and determine the
relationship among different species of organisms. Forensic scientists use DNA to identify suspects using evidence
from crime scenes. Geneticists use DNA to determine the likelihood of an individual or their children to be
susceptible to certain diseases. The field of gene therapy examines DNA to identify the causes of diseases and
develop treatments and cures for the people suffering from these diseases. In today’s lab, you will look at the
structure of DNA, how it is replicated, and how it controls the productions of proteins in the cell.
EXERCISE 1: Human Cheek Cell & Strawberry DNA Isolation (1 point)
Materials:
Ziplock bags
Cheesecloth
Funnel
Ice cold 95% ethanol
Wooden applicator
Test tube
Test tube rack
DNA extraction buffer (100 ml shampoo, 15 g NaCl, 900ml Di-water)
Strawberry (strawberry experiment only)
Drinking water (cheek cell experiment only)
Small drinking cups (cheek cell experiment only)
42
Strawberry DNA Isolation
Procedure (work in groups of 2):
1. Set up the filtration system by placing the funnel in the mouth of the test tube and placing two layers of
cheese cloth over the funnel.
2. Remove stem and leaves from the strawberry 9if necessary). Place the berry in a Ziploc bag and seal it
shut. Mash up the berry for 2 minutes.
3. Add 10 ml of DNA extraction buffer (which is mainly shampoo!) to the berry mush. Continue to mash
the berry and buffer mix for another minute.
4. Pour the berry/buffer mixture into the funnel and let fluid drip into the tube until the tube is about 1/8
full.
5. Slowly pour ethanol down the inside of the tube until the tube is about ½ full.
6. You should see the strands of strawberry DNA floating in the alcohol. Dip the stick into the tube and
twirl and lift stick to spool DNA strands. Observe the texture.
7. STRAWBERRY DNA ISOLATION CLEANUP
a. RINSE OUT THE TEST TUBE AND RE-USE IT FOR THE CHEEK CELL DNA ISOLATION
EXPERIMENT.
Human Cheek Cell DNA Isolation
Procedure (work individually):
1. Add 2 ml of DNA extraction buffer to the test tube. Set the test tube aside in a test tube rack.
2. Pour 10 ml of fresh tap water into a clean drinking cup. Put the 10 ml of water in your mouth and swirl
it around for 2 minutes. Eject the solution back into the cup.
3. Pour about half of the “cheek cell” water into the test tube.
4. Stopper the test tube (with your thumb) and mix the content by gently inverting the test tube several
times.
5. Add approximately 5 ml of ice cold 95% ethanol to your test tube.
6. You should see the strands of your DNA floating in the alcohol. Dip the stick into the tube and twirl
and lift stick to spool DNA strands. Observe the texture.
7. CHEEK CELL DNA ISOLATION CLEANUP
a. THROW DRINKING CUPS IN THE TRASH.

b. RINSE THE TEST TUBE THOROUGHLY WITH WATER AND PLACE IT UPSIDE DOWN IN THE
TEST TUBE DRYING RACK.
43
Questions:
1. Did you notice any visible similarities between the isolated human DNA (your DNA) and the
strawberry DNA? Explain.
2. To isolate DNA (strawberry or human), why did you use a detergent?
EXERCISE 2: DNA is the Molecular Fingerprint (1 point)

Since no two individuals have the exact same nucleotide sequence in their DNA molecules (unless they are
identical twins), biologists can compare DNA of different people by detecting the difference in sequences of their
DNA and distinguish between them. Over 99.9 % of human DNA is the same. DNA profiling focuses on the regions
of the human DNA that vary from one human to the next. These variable segments are different in size (number of
nucleotides) in different individuals. Biologists use gel electrophoresis to detect such differences. The first step in
DNA profiling is to cleave the DNA with enzymes, creating variable DNA fragments. A dye and a fluorescent
substance are added to the DNA fragments and then the DNA is then loaded into a well in a thick, viscous gel. There
are several wells in the gel that contain DNA samples from different individuals. An electric current is then applied
across the gel. Since DNA is negatively charged (due to the phosphate groups of the nucleotides), the fragments
move towards the positive charge, migrating down the gel. The larger fragments move slower than the smaller ones.
After a set time, the electrical current is turned off and biologists can identify DNA fragments that differ in length.
DNA profiling allows scientists to identify criminals, victims, and/or unknown individuals (such as victims of the
2005 Hurricane Katrina). DNA profiling may also used to determine paternity and variety of diseases.
Materials:
DNA profile micrographs
Procedure:
Observe the gel electrophoresis set up and the 2 DNA profile micrographs and answer the following questions.
Questions:
1. Consider DNA profile in Figure 1. Which suspect committed the crime?
2. Consider DNA profile in Figure 2. Which male is the father of the child in this paternity case?
EXERCISE 3: The Structure of DNA (2 points)
DNA is a double-stranded molecule, composed of two nucleotide chains oriented in opposite directions.
Each nucleotide in a chain has a deoxyribose sugar and a phosphate that form the outside of the molecule (called the
backbone), and a base facing the inside or the middle. The bases of one strand pair up with the bases of the other
44
strand to hold the two strands together. In DNA molecules, the base adenine always pairs with the base thymine, and
the base cytosine always pairs with the base guanine. These complementary base pairs are held together by
hydrogen bonds (two hydrogen bonds hold together the A-T pairs and three hydrogen bonds hold together the C-G
pairs). This double-stranded molecule is twisted to form a double helix.
Materials:
K’nex kit and instruction manual
Procedure:
1. Using the K’nex pieces, build a strand of DNA with the sequence 3’ TACCGAGGCATT 5’
2. What will the complementary sequence be? Write it below and build the complementary strand of
DNA using the K’nex pieces.
3. Attach the two strands using the appropriate hydrogen bonds.
4. Holding both ends of the strand, gently twist both ends in opposite directions to observe the double-
helical structure of DNA.
5. After you show the structure of DNA (individually) to your instructor, have the instructor
initial here. ____
EXERCISE 4: DNA Replication (2 points)

DNA must be replicated prior to cell division. DNA REPLICATION can be divided into three steps:
separation, complementary base pairing, and polymerization. During separation the two strands of the double helix
unwind and separate from each other. Then the unpaired bases form new hydrogen bonds with free nucleotides in
the nucleus. These bonds are formed in a complementary fashion: adenine only pairs with thymine and a cytosine
only pairs with guanine. Finally, newly paired bases polymerize (join together) form two new double helices, each
with an original parent strand and a newly formed daughter strand. This type of replication is termed
semiconservative replication.
Materials:
DNA molecule from Exercise 3
Procedure:
1. Separate the two DNA strands from Exercise 3 by breaking the hydrogen bonds one at a time.
2. Add new bases to each parent strand. Be sure to grow each new strand in the 5’ to 3’ direction.
3. Continue breaking the parent strand of DNA and adding new bases until you have two identical DNA
molecules, each one with one old strand and one newly assembled strand of nucleotides. (Note that we
have replicated a strand of DNA of only 12 nucleotide bases. In actual replication, the entire
chromosome is much longer.)
4. After you show replication (individually) to your instructor, have the instructor initial
here. ____
45
5. In the space below, draw the two identical DNA molecules that you have produced. Use different
colors to indicate which strands are from the parent molecule and which are daughter strands.
EXERCISE 5: Transcription (2 points)

The information contained within DNA molecules specifies protein structures. For DNA to direct the
synthesis of new proteins, a transcript of the instructions must be made. This transcript comes in the form of
MESSENGER RNA (mRNA). The single-stranded RNA molecule consists of bases containing a sugar (ribose), a
phosphate, and a nucleotide base. Like DNA, RNA bases include adenine, cytosine, and guanine. Instead of the base
thymine, however, RNA uses the base uracil. In TRANSCRIPTION, the DNA strands separate in the region of the
gene being transcribed. Complementary bases pair up and the nucleotides join to form the mRNA transcript.
Materials:
DNA molecule from Exercise 3 or 4
Procedure:
1. Using one of the DNA molecules you have created, transcribe the 3’ to 5’ strand into an mRNA
molecule.
2. After you show transcription (individually) to your instructor, have the instructor initial
here. ____
Questions:
1. What is the sequence of the DNA molecule you transcribed?
2. What is the sequence of the newly formed mRNA molecule?
46
3. List three differences between DNA and RNA.
DNA RNA
1.
2.
3.
4. List 4 differences between transcription and replication (refer to page 218 of the textbook and the
lecture notes for help).
Transcription Replication
1.
2.
3.
4.
EXERCISE 6: Translation (2 points)

Once the protein-making instructions contained within DNA are transcribed into an mRNA molecule,
TRANSLATION takes place. The mRNA leaves the nucleus and goes into the cytoplasm. Translation takes place
at a ribosome composed of protein and ribosomal RNA. As the mRNA feeds through the ribosome, TRANSFER
RNA (tRNA) brings the appropriate amino acids to the growing polypeptide chain.
47
Materials:
mRNA molecule from Exercise 5
Procedure:
1. Using the mRNA molecule you created in Exercise 5 and the chart below, determine the amino acid
sequence of your new protein.
Questions:
1. List the four codons of your mRNA molecule.
2. For how many amino acids do they code? Which are the coded amino acids?
3. What is the significance of the codon AUG?
4. What sequence do we need at the end of our mRNA for the polypeptide to terminate its growth?
48
MITOSIS AND MEIOSIS
The most remarkable characteristic of cells is that they have the capacity to reproduce themselves exactly.
They can do so because of the unique properties of DNA. The DNA within each nucleus is comprised of several
very long strands, each of which is called a chromosome. The number of chromosomes differs between species, but
is constant within a species; the fruit fly has 4 chromosomes, humans have 46 chromosomes, and some protozoans
and plants have over 200 chromosomes. Not only is the number of chromosomes constant within a species, it is also
constant in the somatic cells (body cells) within an organism. Gametes or sex cells (eggs and sperm) within an
organism contain half the number of chromosomes present in the somatic cells.
Eukaryotic cell division involves nuclear division as well as cytoplasmic division (cytokinesis). There are
two types of eukaryotic nuclear division mechanisms: mitosis and meiosis. Most somatic cells go through
MITOSIS and cytokinesis. The outcome of mitosis of a single diploid cell is two diploid daughter cells which are
clones (genetically identical). Mitosis and cytokinesis are used for growth (of a single fertilized egg into a
multicelluar adult organism), repair (replacement of lost cells), and asexual reproduction. Some somatic cells called
germ cells (located in the reproductive organs of males and females) go through MEIOSIS and cytokinesis. The
outcome of meiosis of a single diploid cell is four haploid gametes which are genetically different from each other.
The only function of meiosis and cytokinesis is gamete production. Gametes( sperm and egg) are used in sexual
reproduction. The goal of this lab is to acquaint you with the structure of chromosomes and the two types of cell
division.
THE CELL CYCLE

A majority of the time our chromosomes exist within the nucleus as strands so long and thin that they are
invisible to the light microscope. During this period of time, termed INTERPHASE, both the production of proteins
and the replication of the DNA itself are occurring. It is important that the DNA within the nucleus is replicated
exactly to provide a full complement of DNA for both of the daughter cells that will result from nuclear division
(mitosis or meiosis) and cytokinesis. After interphase is complete, the DNA strands become so highly coiled that
they become visible in the light microscope; this is what we see as “chromosomes.” At this point in the cell cycle,
termed PROPHASE, each chromosome is composed of two identical copies (sister chromatids) attached to one
another by a structure called a centromere. During the next few phases (METAPHASE, ANAPHASE, and
TELOPHASE) the chromosomes go through an orderly sequence of movements to ultimately endow each of the
two resulting daughter cells with the appropriate chromosome content. Since the chromosomal content of somatic
cells is different from that of gametes, the movements of the chromosomes differs slightly (but importantly) between
mitosis and meiosis. It is the exact pattern of chromosomal movement during both mitosis and meiosis that we are
going to examine in this lab.
MITOSIS
Mitosis involves a single nuclear division; the process is one of continuous change, not a series of discrete
steps. However, for simplicity we usually identify and name several phases, which are key intermediate
chromosomal arrangements in this continuous process. The first is prophase, during which the chromosomes start to
condense and become visible within the cell, often appearing as a coiled mass (although the chromosomes exist as
pairs of sister chromatids at this point, they are much too small for you to tell). During metaphase, the chromosomes
line up near the center of the cell. At this point, very thin hollow tubes called spindle fibers attach to the centromeres
of the chromosomes, and will later pull them to their respective sides of the cell. Metaphase is followed by
anaphase, during which the sister chromatids are pulled apart to opposite sides (“poles”) of the cell. Telophase
completes the nuclear division process; as the chromosomes have reached the opposite ends of the cell, they
decondense and a nuclear envelope forms around them.
49
Cytokinesis by furrowing (in animal cells) or by cell plate formation ( in plant cells) starts at the end of
anaphase and is completed before telophase is finished. Each one of the two genetically identical daughter cells now
return to interphase, during which protein synthesis and DNA replication will occur. Ultimately they will once again
divide by mitosis and cytokinesis to form two daughter cells each.
EXERCISE 1: Mitosis Simulation with Beads (1.5 points)

In order to more fully understand the arrangements and movements of chromosomes during mitosis, we
will simulate chromosomes with snap beads; this is perhaps the best way to really see how much you know about
mitosis.
Materials:
Four 4” lengths of snap beads (2 each of 2 colors)
Mitosis poster templates
Procedure:
1. Obtain four 4” lengths of snap beads. Make sure that two are made of beads of one color, and two are
made of beads of a different color. These will represent chromosome pair #1
made of beads of a different color. These will represent chromosome pair #2.
3. Using the Mitosis template, place the four pairs of “sister chromatids” inside one circle in a prophase
arrangement (a random pile).
4. First as a group, then as individuals, practice moving and arranging the chromosomes in the different
configurations of mitosis. Be sure to use and practice the correct terminology (sister chromatids,
homologous chromosomes, etc). Call your instructor for help if you have difficulty.
5. After each set of divisions, check to make sure the daughter cells are identical to the parent cell, and
then repeat the exercise.
6. After demonstrating (individually) mitosis to your instructor, have him/her initial here. ____
EXERCISE 2: Onion Root Tip Mitosis (2 points)
Plants such as the onion prove to be excellent specimens for viewing the process of mitosis for several
reasons. First, the chromosomes are rather large and few in number, which makes their identification much easier.
Second, the cells are held in place by a rigid cell wall, and large numbers of neighboring cells are visible at one time.
Third, there are discrete regions within the root where there is quite a high rate of mitosis, so actively dividing cells
in all stages are easier to find.
Materials:
Prepared slide of onion root tip
Procedure:
1. Examine the cells comprising the onion root tip, approximately ⅓ of the distance from the dark-
staining tip. You will need to first find this region of cell division under low power, then increase the
magnification to 400X . Note the cells in various stages of mitosis (they look similar to the photos in
your text book). Draw the chromosomes in each of the mitotic stages below.
50
Interphase Prophase Metaphase Anaphase Telophase
2. Once you have familiarized yourself with some of the stages of mitosis, count 50 cells at random (all
within one or two fields of view) and determine how many cells are in each of the stages of mitosis.
Put your data on the board, combined with the data from the rest of the class, and determine what
percentage of the cells in the region of cell division is in each mitotic stage. The preparation of the
slide represents a sort of “snapshot in time” of the cellular activities; the percentage in each stage
relates roughly to the time a single cell spends in each stage of division.
Stage of Cell Division Your Data Class Data (Number of Class Data
(Number of Cells) Cells) (% of Cells)
Interphase
Prophase
Metaphase
Anaphase
Telophase
Total
Questions:
Use the above table and answer the following questions.
1. About what percentage of the time are cells in:

a. Interphase ________
b. Prophase ________
c. Metaphase ________
d. Anaphase ________
e. Telophase ________
2. If the entire cell cycle takes about 24 hours, how long do cells stay in interphase?
3. Why do you think cells spend most of their time in interphase?
51
MEIOSIS
While mitosis and cytokinesis result in the production of somatic cells, GAMETES, or sex cells, are
produced by meiosis and cytokinesis. Meiosis occurs only in the germ cells located in the ovaries and testes, and
leads to the production of eggs or sperm. The unique property of meiosis is that it reduces the chromosome content
of the daughter cells to exactly one half of that of the parent cell (from DIPLOID to HAPLOID). The alignments
and movements of chromosomes in meiosis separate HOMOLOGOUS CHROMOSOMES, so that each resultant
cell receives one copy (out of two) of each of the homologous chromosomes.
The mechanics of the process of meiosis are very similar to that of mitosis, with a few important changes.
First, meiosis involves two sets of divisions. In the first set of divisions (Meiosis I), homologous chromosomes lie
next to each other during Metaphase I, and are pulled to opposite poles during Anaphase I. The second set of
divisions (Meiosis II) is very similar to mitosis, with the chromosomes arranged along the equator during Metaphase
II, and sister chromatids pulled to opposite poles during Anaphase II. Thus the first set of divisions serves to
separate homologous chromosomes, and the second set separates sister chromatids, resulting in four genetically
different daughter cells each containing half of the genetic information contained in the parent cell. In addition to
reduction of the number of chromosomes in half, two other interesting events take place during Meiosis I. These
events are crossing over of the homologous chromosomes or GENETIC RECOMBINATION in Prophase I and
the random alignment of the homologous chromosomes in Metaphase I. Both of these events contribute to the
genetic variation in the offspring. A third factor that contributes to the genetic variation in the offspring occurs
during fertilization. At fertilization, of all the genetically diverse gametes produced, chance will determine which
two will meet.
EXERCISE 3: Karyotype of Human Chromosomes (0.5 points)

Chromosomes are visualized in a lab preparation called a karyotype. Each chromosome has distinct size,
length, centromere location, and banding (staining) patterns.
Materials:
Karyotype of human chromosomes
Procedure:
1. Examine the two karyotype preparations of the human chromosomes at the side table. Answer the
following questions.
Questions:
Karyotype # 1
1. In the above karyotype, were the human chromosomes prepared before or after the S phase of the
interphase? How can you tell?
2. How many pairs of human chromosomes are present?
3. The two chromosomes in each pair are referred to as?
52
Karyotype # 2
4. Does this karyotype indicate a male or a female human?
5. Does this karyotype reveal an abnormality? Explain.
EXERCISE 4: Meiosis ( 2 points)

Materials:
Meiosis poster templates
Procedure:
made of beads of a different color.
made of beads of a different color.
3. Using the Meiosis template, place four pairs of “sister chromatids” inside one cell in a prophase
arrangement (a random pile).
4. Mark the maternal and parental chromosomes with a single different colored bead at both ends of the
sister chromatids (for example, put a single white bead on the end of all of the paternal chromatids, and
an orange bead on the end of all the maternal chromatids).
5. During Prophase I, place the homologous chromosomes side-by- side, and exchange equal lengths of
chromosomes between them (now the sister chromatids should appear different from each other).
Draw a “normal” and a “recombined” chromosome below.
6. First as a group, then as individuals, practice moving and aligning the homologous chromosomes in
different arrangements in metaphase I. Note that random alignment of the homologous chromosomes
leads to gametes with different genetic information.
7. Carry out Meiosis I and Meiosis II. Be sure to use and practice the correct terminology (sister
chromatids, homologous chromosomes, etc.). Call your instructor for help if you have difficulty.
8. After each set of divisions, check to make sure you have the correct types and numbers of
chromosomes.
9. After you demonstrate Meiosis (individually) to your instructor, have them initial here. ____
53
10. When you complete the set of divisions, take the newly formed “gametes” and combine two to form a
diploid offspring. Now go through several sets of division by mitosis, and then one of meiosis, with the
chromosomes of this new individual.
Questions:
1. Why is it important that homologous chromosomes pair correctly during meiosis? Why can
homologous chromosomes line up in any order during mitosis?
2. In the above exercise, how many different types of gametes can be produced from the original parent
cell?
3. Humans have 46 chromosomes (23 pairs of homologous chromosomes).
a. How many chromosomes are there in human gametes?
b. How many in a human zygote?
4. What can you say about the genes on the recombined chromosomes with respect to maternal and
paternal origin (are the resulting chromosomes composed completely of maternal genes or paternal
genes)?
5. How does meiosis and cytokinesis differ from mitosis and cytokinesis (give four differences) ? What is
significant about the differences?
EXERCISE 5: Independent Assortment of Alleles (2 points)

In the previous exercise you examined the process of gamete formation. You found out that meiosis
reduces the number of the chromosomes (in the parental germ cell) by half. During anaphase I, homologous
chromosomes are separated from each other and only one of them ends up in each of the resulted gametes. If you
consider the fact that homologous chromosomes contain homologous alleles for particular traits, then we can say
that during anaphase I, homologous alleles separate from each other and only one of them will end up in each of the
up coming gametes. The separation of the homologous alleles located on one pair of homologous chromosomes and
the distribution of these alleles into the up coming gametes is done independently of the separation of the
homologous alleles located on another pair of homologous chromosomes (this is Mendel’s second law that we will
discuss in Ch. 11 and in your next lab). This independent assortment of the homologous alleles could give rise to
54
many possible combinations of maternal and paternal alleles in the resulting gametes. In this exercise you will
examine the process of independent assortment of the homologous alleles in the resulting gametes.
Materials:
Popsicle sticks (6 per student)
Sharpie markers (1 per student)
Procedure:
1. Give your personal information about the following traits in the table below. If you have a dominant
phenotype, assume that you have the heterozygous genotype for that trait.
a. Widow’s Peak (pointed hairline). If present your genotype is WW or Ww. If not present your
genotype is ww.
b. Pigmented iris. Green, hazel, brown, or black eyes due to genotypes PP or Pp. Blue or gray
(pp).
c. Tongue-Rolling. If you can roll your tongue into a U-shape your genotype is RR or Rr. If you
can not roll your tongue into a U-shape, your genotype is rr.
d. Attached earlobes . If present your genotype is aa. If not present your genotype is Aa or AA.
e. Dimples. If present your genotype is DD or Dd. If not present your genotype is dd.
Trait Your Phenotype Your Genotype
Widow’s Peak
Pigmented Iris
Tongue-Rolling
Attached Earlobes
Dimples
2. Obtain 6 popsicle sticks. Using a Sharpie, write your initials on the top of both sides of every stick.
Each stick represents one pair of homologous chromosomes. One stick will represent the sex
chromosomes. Each side of the stick represents one chromosome of a homologous pair.
3. Using the table above write your two alleles for each trait on one stick. Write one allele in the middle
of the stick on one side and the other allele in the middle of the stick on the other side. Use one stick
per trait.
4. The last stick will represent the sex chromosomes. Write an X on both sides of the stick (in the middle)
if you are a female. Write an X on one side of the stick (in the middle) and a Y on the other side (in the
middle) if you are a male.
5. Now you can show the process of independent assortment of the homologous alleles by holding your
bundle of sticks above the table and dropping them. Five alleles and one sex chromosome will end up
on the table. The side of the stick (one of the two alleles) that appears on the table is completely by
chance. List the alleles. _________________________
Do it again. List the alleles. _________________________
Do it again. List the alleles. _________________________
Each list represents the genetic content of one of the many possible gametes that you could make.
55
Questions:
1. How many different combinations do you think are possible with 23 pairs of homologous
chromosomes? How does this affect your ability to predict with any certainty the inheritance of traits
in humans?
2. What does each stick represent?
3. What does each side of a stick represent?
EXERCISE 6: Fertilization (2 points)

In the previous exercises you became familiar with events such as crossing over of the homologous
chromosomes and random alignment of the homologous chromosomes which leads to the independent assortment of
the homologous alleles into the resulting gametes. These two events are two of the major events that contribute to
genetic variation in the offspring. The third event that introduces genetic variation in the offspring is fertilization. At
fertilization, a female and a male gamete come together, each contributing their haploid number of chromosomes to
produce a diploid cell (zygote, the first cell of the offspring). At fertilization, of all the genetically diverse gametes
produced, chance determines which two will meet. In the following exercise you will examine how chance affects
fertilization.
Materials:
Your labeled popsicle sticks (6 per student)
Procedure:
1. Pair up with someone of the opposite sex. Each of you will need your 6 chromosomes (sticks).
2. Drop your sticks together on the table. Dropping the sticks represents a fertilization event.
3. The genetic makeup of the zygote is represented by both sets of alleles showing on the sticks.
4. Record the results in the table below.
56
Child #1
Parents’ Names__________________________________
Child’s Name___________________________________
Mother’s Father’s Child’s

Trait Child’s phenotype
Alleles Alleles Genotype
Widow’s Peak
Pigmented Iris
Tongue-Rolling
Attached Earlobes
Dimples
5. Repeat steps 1-4 to produce child #2 and record the results in the table below.
Child # 2
Parents’ Names________________________________
Child’s Name___________________________________
Mother’s Father’s Child’s

Trait Child’s phenotype
Alleles Alleles Genotype
Widow’s Peak
Pigmented Iris
Tongue-Rolling
Attached Earlobes
Dimples
Questions:
1. Which parent determines the sex of each child?
2. Which traits do the two children have in common?
3. We looked at only five traits. It is estimated that humans possess 20,000-25,000 genes. Why is it
unlikely that any two non-identical siblings will look the same?
57
MENDELIAN GENETICS
GENETICS is the study of hereditary transmission and variation. In exercises conducted in the last lab,
you learned about the movement of chromosomes within a cell during both mitosis and meiosis. However, you
didn’t learn how cells get the particular chromosomes they have, or how those particular chromosomes produce
different “traits” in different organisms. This is what you will work on in the current lab.
Gregor Mendel is considered the “father of genetics”, yet he had no idea of chromosomes, genes or DNA.
He only knew that parents passed on traits to their offspring, and did so in predictable patterns. Thus, you are going
to begin your study of genetics with much more information than Mendel could have hoped for in the late 1800’s.
We now know, for example, that hereditary information is encoded in GENES, sequences of DNA which code for a
single protein, and that CHROMOSOMES are very long strands of DNA containing hundreds or thousands of
genes. We also know that mitosis produces exact copies of diploid cells, while meiosis reduces diploid cells to
haploid cells and results in the production of gametes. Each diploid cell has two copies of every chromosome, one
from the mother and one from the father; these chromosomes are called HOMOLOGOUS CHROMOSOMES,
and the different forms of a gene are called ALLELES.
Even without this wealth of information, Mendel was able to formulate two basic laws concerning
chromosomal arrangements in gametes. The first is the LAW OF SEGREGATION, which states that different
forms of a gene (alleles) move as independent units from one generation to the next (whether or not they produce
visible traits), and that they segregate at random into gametes during meiosis (Anaphase I), thus producing
predictable ratios of traits in the offspring. The second is the LAW OF INDEPENDENT ASSORTMENT, which
states that alleles for different genes segregate and group (assort) independently of each other. Using these two laws,
one can determine 1) the ratio of different gametes any given genotype can produce, and 2) the ratio of offspring
genotypes and phenotypes produced by the cross of any two individuals. These will be illustrated in the following
exercises.
MENDELIAN GENETICS IN CORN

Corn is an excellent species to use for analysis of patterns of inheritance because a single ear of corn is
actually a collection of hundreds of offspring. Thus, if the genotypes of the parents are known, it is possible to easily
observe the ratios of phenotypes in a large number of offspring relatively quickly. The corn we will use for the
following exercises is similar to that which you eat. Several visible (phenotypic) differences exist and we will use
these differences to examine patterns of inheritance. The two phenotypic traits we will observe involve the color and
shape of the kernels. Corn has a protein layer in the seed coat (the aleurone layer), which can be either yellow or
purple. There is a single gene which codes for aleurone protein, with the purple (R) allele dominant to the yellow (r)
allele. Corn also has the capacity to store carbohydrates as starch or simple sugars, and this capacity is also
determined by a single gene with two alleles. Kernels which store carbohydrates as starch (S) have smooth surfaces,
while those which store carbohydrates as simple sugars (s) have a wrinkled surface. The S allele is dominant to the s
allele. In the following exercises you will be provided with ears of corn that represent the F2 generation. The P
generation cross was a doubly homozygous cross, RRSS x rrss. You will need to determine the genotype of the F1
generation.
EXERCISE 1: Monohybrid Cross In Corn (2.5 points)

In this exercise, you will observe the phenotypic frequencies of corn kernels with respect to coat color.
Remember R is the allele for purple seeds and is dominant to the r allele for yellow seeds.
Materials:
Ear of F2 corn
58
Procedure:
1. Use the given P generation genotypes to fill in the P generation cross:
____________ X _____________
2. Use a Punnett square to determine the possible F1 genotypes.
a. What is the predicted genotype ratio among F1?
b. What is the predicted phenotype ratio among F1?
c. Are all of the offspring expected to be the same?
3. Use the data obtained to determine the F1 generation cross:
____________ X ______________
59
b. What is the predicted phenotype ratio among F2? Your predicted phenotypic ratio will serve
as your hypothesis.
5. Test your hypothesis by counting 100 corn kernels at random (count several rows).
Color Number of Seeds
Purple
Yellow
6. Combine your data with the class. This will serve to increase your sample size and reduce the
likelihood that you will count more of one phenotype by chance (this is known as sampling error). If
this does not seem to make sense, think about flipping a coin. You have a 50/50 chance of flipping
heads. If you flip the coin 4 times, you may actually get 4 heads instead of the 2 predicted. If you flip
that same coin 1,000,000, you’re more likely to get 50% heads. You should be a little freaked out if
you flip heads a million times in a row!
Color Number of Seeds Percent of Seeds
Purple
Yellow
Questions:
1. What is the actual phenotype ratio? This is class result.
purple: yellow
2. How does this value compare with the predicted value (from 4b above)?
3. From the given information, can you determine the genotype of any individual kernel? Why or why
not?
60
4. What phenotypes would you expect in the F2 generation if they followed the incomplete
dominance model?
5. What phenotypes would you expect in the F2 generation if they followed the co-dominance
model?
EXERCISE 2: Monohybrid Cross In Corn (2.5 points)

In this exercise, you will observe the phenotypic frequencies of corn kernels with respect to coat texture.
Remember S is the allele for smooth seeds and is dominant to the s allele for wrinkled seeds.
Materials:
Ear of F2 corn
Procedure:
1. Use the given P generation genotypes to fill in the P generation cross:
____________ X _____________
3. Use the data obtained to determine the F1 generation cross:
____________ X ______________
61
as your hypothesis.
5. Test your hypothesis by counting 100 corn kernels at random (count several rows).
Texture Number of Seeds
Smooth
Wrinkled
6. Combine your data with the class. This will serve to increase your sample size and reduce the
likelihood that you will count more of one phenotype by chance (this is known as sampling error). If
this does not seem to make sense, think about flipping a coin. You have a 50/50 chance of flipping
heads. If you flip the coin 4 times, you may actually get 4 heads instead of the 2 predicted. If you flip
that same coin 1,000,000, you’re more likely to get 50% heads. You should be a little freaked out if
you flip heads a million times in a row!
Texture Number of Seeds Percent of Seeds
Smooth
Wrinkled
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Questions:
smooth: wrinkled
not?
4. Which of Mendel’s Laws are you exploiting in the monohybrid cross analysis? Explain.
EXERCISE 3: Dihybrid Cross In Corn (3 points)

In a dihybrid cross we are following the patterns of inheritance of two different traits (genes). In the
following exercise, you will determine how the previous traits, coat color and coat texture, combine in the offspring.
To the uninformed, it might seem like the dominants should always travel together, so that when you have purple
seeds you will also have smooth seeds. This isn’t necessarily the case, however. As long as the two genes reside on
different chromosomes (i.e., are not linked), they should assort (combine) independently of one another in the
offspring (Mendel’s second law). We will test this assumption.
Materials:
Ear of F2 corn
Procedure:
1. Use the given P generation genotypes to fill in the P generation cross (see page 50).
____________ X _____________
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2. Use the data obtained to determine the F1 generation cross.
____________ X ______________
3. Use a Punnett square to determine the possible F2 genotype(s)
as your hypothesis.
4. Test your hypothesis by counting 100 corn kernels at random (count several rows)
Phenotype Number of Seeds
Purple, Smooth
Purple, Wrinkled
Yellow, Smooth
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Yellow, Wrinkled
5. Combine your data with the class.
Phenotype Number of Seeds Percent of Seeds
Purple, Smooth
Purple, Wrinkled
Yellow, Smooth
Yellow, Wrinkled
Questions:
_____Purple, Smooth: _____Purple, Wrinkled: _____Yellow, Smooth: _____Yellow, Wrinkled
not?
4. Which of Mendel’s Laws are you exploiting in the dihybrid cross analysis? Explain.
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EXERCISE 4: Visible Phenotypes in Humans (2 points)

In this exercise you will examine some traits possessed by you and your classmates. The following traits
are all controlled by a single gene. Notice as you proceed through the list that just because a trait is common in a
population does not necessarily mean that it is controlled by the dominant allele.
Materials:
PTC paper
Procedure:
1. Read the descriptions of each trait.
2. Fill in your phenotype on the chart.
3. If possible, list your genotype in the chart. For example, if you have the recessive trait for gene G, your
genotype is homozygous recessive gg. If you have the dominant trait for gene G, your genotype is not
fully known, so fill in G_ on the chart. If you know that one of your parents is recessive for that trait,
you must have received a recessive allele (g) from him/her, so your genotype is Gg.
4. Give your phenotype to your instructor so he/she can provide you with the phenotypic results for the
class.
Widow’s peak The W allele for a widow’s peak (pointed hairline) is dominant to the w allele for the
straight hairline.
Bent little finger The B allele for a pointed little finger is dominant to the b allele for a straight little finger.
You have the bent little finger if you lay your hand flat on the bench and the last joint of
your little finger bends toward your fourth finger.
Pigmented iris The P allele produces pigment in the front layer of the iris, resulting in green, hazel,
brown, or black eyes. This is dominant to the p allele, which does not produce pigment in
the front layer, and results is blue or gray eyes (you are seeing the color of the back layer
of iris).
Attached earlobes The A allele for free earlobes is dominant to the a allele for attached earlobes.
Hitchhiker’s thumb The H allele for non-hitchhiker’s thumb is dominant to the h allele for the hitchhiker’s
thumb (you can bend your thumb back at an angle of 60° or more at the last joint.
Interlacing fingers Fold your hands together with your fingers interlaced. If your left thumb crosses over the
right, you have the dominant allele C. The c allele is recessive, with the right thumb
crossing over the left.
PTC tasting Chew on a piece of PTC paper (phenylthiocarbamide). If you detect a bitter taste, you
have the dominant T allele. The t allele is recessive and results in no taste detection.
Mid-digital hair The M allele for hair on the middle segment of your fingers is dominant to the m allele
for no hair.
Polydactyly The Y allele for extra digits on a hand or foot is dominant to the y for five fingers on each
hand and five toes on each foot
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# of Dominants in # of Recessives in
Characteristic Your Phenotype Your genotype
class class
Widow’s peak
Bent little finger
Pigmented iris
Attached earlobes
Hitchhiker’s thumb
Interlacing fingers
PTC tasting
Mid-digital hair
Polydactyly
Questions:
1. Are dominant characteristics always more frequent in a population than recessive characteristics? Why
or why not?
2. Which, if any, recessive characteristics occur more frequently in your lab population?
3. Is it possible to determine the genotype of individuals having the dominant phenotype? How?
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ORGAN SYSTEMS
All organisms must exchange materials between their cells and the environment; this exchange includes
both the uptake of materials necessary to sustain life (such as oxygen and nutrients) and the elimination of metabolic
wastes which are toxic (such as carbon dioxide and urea). In large multicelluar organisms such as humans, this
process is extremely complex, as literally trillions of cells must have their uptake and elimination needs met on a
continual basis, 24 hours a day, every day of their life.
The systems we will study in the following lab are responsible for a large percentage of the uptake and
elimination just discussed. We will examine: 1) the CIRCULATORY SYSTEM, which is specialized for moving
materials around within the body, 2) the RESPIRATORY SYSTEM, which is specialized for exchanging gasses
(mainly O2 and CO2) with the environment, 3) the DIGESTIVE SYSTEM, which is specialized for removing the
nutrients from food and absorbing them into the body, and 4) the URINARY SYSTEM, which is specialized for
filtering your blood and removing wastes (salts, urea, etc) and returning “good” materials into circulation.
THE CIRCULATORY SYSTEM

Circulatory systems have three basic components: a circulating fluid (the BLOOD), tubes which contain
and direct the flow of the fluid (ARTERIES, VEINS, and CAPILLARIES), and a pump to keep the fluid
continuously circulating (the HEART). In the current lab, we will: 1) identify the components of blood, and become
familiar with their functions, 2) observe arteries, veins and capillaries, 3) identify the major structures of the
mammalian heart, and know the function of each structure, 4) measure the heart rate, blood pressure and electrical
activity of a human heart, and 5) identify the major structures of the circulatory system in a dissected mammal
BLOOD COMPOSITION
Blood is composed of a large number of cells suspended in a watery solution. The majority of the cells are
RED BLOOD CELLS (RBCs), which are specialized for carrying oxygen. There are over 1 million RBCs per drop
of blood! A minority of cells, the WHITE BLOOD CELLS (WBCs), is involved in the immune system; there are
approximately 500-1000 RBCs for every one WBC. Both the fluid volume of blood and the number of WBCs and
RBCs are maintained relatively constant in humans; a deviation often indicates some sort of physiological problem,
so a screening of the blood is a common first-step in the diagnosis of diseases. One common clinical test is a
determination of the hematocrit, or the fraction of the blood composed of cells (about 45%) and that composed of
watery plasma (about 55%). This test is accomplished by spinning (in a centrifuge) a sample of whole blood until
the denser cells settle out, leaving a yellowish liquid (the plasma) above. Another common clinical test is the routine
scanning of the blood cells themselves to detect actual numbers of different types of cells in the cellular portion of
blood; an excess or deficiency of RBCs or WBCs is an indication of certain physiological problems.
EXERCISE 1: Blood Composition (1 point)

Materials:
Hematocrit tube (on display)
Prepared slide of human blood
Prepared slide of sickle-cell anemia
Micrographs of various blood pathologies
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Procedure:
1. Observe the sample of centrifuged sheep’s blood on display (do not shake or tip the tube).
packed cell volume (mm)

Hematocrit = X 100
total blood volume (mm)
2. Obtain a prepared slide of human blood. View the blood cells at high power (400X). Note the
numerous small, pinkish cells (RBCs) and the occasional, larger purple-stained cells (WBCs). You will
have to move the slide around quite a bit to see very many WBCs. Draw the RBCs and several
different WBCs.
Red blood cells White blood cells
2. Obtain a prepared slide of human sickle-cell anemia. View the blood cells at high power and draw
them below. Note how the diseased cells differ from normal human blood cells.
Sickle-cell anemia
3. Examine a few of the micrographs showing other human blood pathologies. Note how the blood in
these micrographs differs from normal human blood.
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Questions:
1. Do all of the WBCs look alike? Look at the different types of WBCs shown in your textbook, and see
if you can identify any of the different types on your slide. Label them in your drawing above.
2. How do the RBCs in individuals with sickle-cell anemia differ from normal human RBCs? Why is this
problematic for individuals that have sickle-cell anemia?
BLOOD VESSELS
In the early 1600s, scientists first produced experimental evidence that blood flows away from the heart in
arteries, and towards the heart in veins. It was soon discovered that these two types of vessels were connected to
each other by even smaller vessels, the capillaries. Thus the blood flows through the body in series of vessels
together called the circulatory system: away from the heart in arteries, through tissues and organs in dense beds of
capillaries, and eventually back to the heart in veins. In an undamaged circulatory system, RBCs never leave the
vessels, but plasma may “leak” in or out through the thin capillary walls. In the following exercises, you will
examine prepared slides of two types of blood vessels, and observe blood flowing through capillaries in the tail of a
goldfish.
EXERCISE 2: Observation of Blood Vessels (0.5 point)

The three types of blood vessels can be distinguished by their size and thickness: arteries are thick-walled
and rather elastic, veins have much thinner and less elastic walls, and capillaries are extremely small (in fact, even
the largest capillary is too small to be seen by the naked eye). In the following exercise you will examine a prepared
microscope slide of an artery and vein.
Materials:
Prepared slide of blood vessels
Procedure:
1. Examine and draw the arteries and veins (from the slide) and capillaries (from the gold fish).
Artery Vein Capillaries
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Questions:
1. Can you distinguish between the arteries and veins based on the thickness of the walls? How?
EXERCISE 3: Observation of Blood Flow in Capillaries (1.5 point)

Capillaries are too small to be seen by the unaided eye, and are usually buried too deep within living tissue
to be easily observed “in action.” However, the skin present within the tail fin of a goldfish is transparent enough so
that blood flow in the capillaries can be viewed quite easily.
Materials:
Petri dish (top or bottom half)
6” square of gauze
Ice bath
Microscope slide
Goldfish
Procedure:
Note: To avoid harming the goldfish, this exercise will be set up by your instructor.
1. Remove the goldfish from the aquarium.
2. Wrap the fish (except head and tail) with dripping wet, chilled gauze.
3. Place the wrapped fish in the Petri dish. Place the microscope slide over the thin (terminal) part of the
tail.
4. Place the Petri dish on the microscope, and examine under low, then high power.
5. Observe blood cells moving through the capillaries
Questions:
1. Is the movement of blood cells constant or do they “pulse” through the vessels?
2. Relative to the size of the blood cells, about how large are the capillaries?
3. Relative to arteries and veins, how big are capillaries?
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4. Based on the differences in wall thickness between arteries and veins and capillaries, can you explain
why only capillaries are “leaky”?
THE MAMMALIAN HEART

The mammalian heart is a pump constructed of muscular walls surrounding 4 separate (but connected)
chambers. When the muscles contract the volume of these chambers is reduced, hence the internal pressure increases
and blood is propelled out under rather high pressure (remember, when blood leaves the heart it is traveling within
arteries). The heart can be thought of as essentially two sets of pumps lying side-by-side; each set has an uppermost
chamber called the ATRIUM (plural atria), which has rather thin muscular walls and serves only to collect blood
returning to the heart (within the veins) and pump it into the chamber below. Below each atrium is a chamber called
the VENTRICLE, which has much thicker muscular walls and serves to pump blood out of the heart. The ventricles
are the part of the heart that usually comes to mind when most people think of a heart.
A system of valves prevents backflow of blood from the ventricles into the atrium: the BICUSPID
VALVE (or mitral or left atrioventricular valve) on the left side and the TRICUSPID VALVE (or right
atrioventricular valve) on the right side. The “loose” ends of the valves are held in place (i.e., prevented from
collapsing) by several very strong strands of connective tissue called CHORDAE TENDINEAE, that are connected
to the ventricle walls via small mounds of cardiac muscle called PAPILLARY MUSCLES.
The set of chambers (atrium and ventricle) on the left side of the heart pumps blood out to all parts of the
body except the lungs; this circulatory pathway is called SYSTEMIC CIRCULATION. Blood in systemic
circulation flows to the brain, eyeballs, nose, mouth, arms, legs, digestive organs, skin, and to the heart muscle itself.
Since the left ventricle must pump blood up to the brain, out to the fingers, to all the digestive organs, and down to
the toes, quite a bit of force is generated by the contraction of the left ventricle. The set of chambers of the right side
of the heart pumps blood out to the lungs; this pathway is called PULMONARY CIRCULATION. Because the
heart is situated in the chest only inches from the lungs, little force is necessary to pump blood to these organs.
EXERCISE 4: Anatomy of the Mammalian Heart (1 point)

Materials:
Sheep hearts cut in several planes or plastic model
Gloves
Blunt probe
Procedure:
1. Obtain a heart from the specimens provided. If it has been cut into more than one piece, attempt to
view all of the pieces at once.
2. Locate and identify the following structures:

 Right and left ventricles
 Right and left atria
 Bicuspid and tricuspid valves
 Chordae tendonae
 Papillary muscles
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3. Using your finger or a blunt probe as a guide, follow the path of blood flow into and out of the four
chambers, identifying the names of the vessels (systemic artery and vein, pulmonary artery and vein)
connected to each chamber.
Questions:
1. Name the functions of the following parts of the heart.
Atria:
Ventricles:
Valves:
Cordae tendineae:
Papillary muscles:
2. Trace the path of blood flow into and out of the heart’s four chambers.
3. Explain the different thicknesses of the walls of the chambers based on their functions.
THE HUMAN HEART

Your heart is very similar to the ones you have been looking at (maybe a bit larger than the sheep heart), so
we will not be viewing human hearts. We will, however, be examining the functions of living human hearts. As you
are conducting the following exercises, keep in mind what you have just learned about blood and the heart.
The rhythmic beat of the heart begins within a few specialized cells called the pacemaker. Although it may
sound and feel like a single contraction, a wave of contraction spreads across the heart: the atria contract first to fill
the ventricles, then the ventricles contract to force blood out to the arteries. Of course, as the ventricles contract they
create a rather high internal pressure in the arteries, and when the ventricles are relaxed the arterial pressure
decreases a bit. These two pressure states of the arteries are called systolic (ventricular contraction phase) and
diastolic (ventricular relaxation phase), and are the two numbers given when your blood pressure is read
(systolic/diastolic).
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EXERCISE 5: Human Heart Sounds (0.5 points)
The heart sounds are best detected using an instrument called a stethoscope; they are usually described as a
“Lub” (systolic) and “Dup” (diastolic) sound. The “Lub” sound is the valves between the atrium and ventricle
vibrating as the ventricle contracts, and the “Dup” sound is that of the valves between the arteries and ventricle
vibrating as the ventricle relaxes.
Materials:
Stethoscope
Alcohol wipes
Procedure:
1. Clean and disinfect the earpieces of a stethoscope with an alcohol wipe. Place the earpieces of the
stethoscope comfortably in your ears.
2. Place the end of the stethoscope on the chest over the area of the heart (yours or your lab partner’s).
Listen.
3. While listening to the heartbeat, try to feel the carotid (neck) or wrist pulse of the volunteer to correlate
the sounds to systole and diastole.
4. Move the stethoscope to various regions of the chest (corresponding to different portions of the heart),
and listen.
5. Count the number of heartbeats in a minute. Have the volunteer climb up and down the stairs several
times, and measure the pulse rate again.
Beats per minute resting = Active =
Questions:
1. Did the heartbeat sound different in different parts of the chest? During diastole and systole?
2. Why did the pulse rate change after exercise?
EXERCISE 6: Mammalian Circulatory System (1 point)
Blood must be transported to and from every cell in the body on a continuous basis. To accomplish the
necessary distribution of blood, mammals have a series of interconnecting vessels (arteries, capillaries and veins)
which directs the flow of blood in a very organized manner (systemic circuit). Every organ and tissue in the body is
supplied with an artery, a vein and a capillary bed.
In the following exercise you will observe a dissected mammal to get a feeling for the complexity of the
circulatory system. The animals have been injected with latex (rubber) to help you identify the different blood
vessels; arteries appear red and veins appear blue.
Materials:
Preserved animal on display (cat or fetal pig)
Gloves (if you are going to touch the animals)
Blunt probes
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Procedure:
1. In this exercise, you are to examine the heart and blood vessels of one (or more) of the animals on
display. Refer to the manual and/or handouts to aid in identification of vessels. Do not worry about
learning all the names of the vessels; rather, be able to distinguish veins from arteries and to determine
the direction of blood flow.
2. Begin by finding the systemic artery leading away from the heart; it has branches which go up to the
head, out to the arms, and down along the back. Follow some of these branches (especially the one
along the back), and not the smaller arteries branching off at various points. Again, follow some of
these smaller branches to their destinations (such as the stomach, the spleen, and the intestines). Note
the presence of veins (sometimes injected with yellow latex in the digestive organs) originating from
these organs. Follow the path of blood flow through the veins, eventually winding up back at the heart.
Questions:
1. Trace the flow of blood from the heart, through the organs, and back to the heart.
2. How many arteries and veins flow into and out of each organ?
3. Which organs seem to have the largest arteries? The most arteries? Why do you think this is so?
THE RESPIRATORY SYSTEM

Organisms which respire aerobically must provide a continuous supply of oxygen and high energy organic
molecules (“fuels”) to their cells. Because the “burning” of these fuels produces byproducts which are toxic at high
concentrations, bodies must also continually remove the resulting wastes from the cells. The respiratory system is
involved with the gain and elimination of the gases involved in aerobic respiration -- oxygen and carbon dioxide.
A point to remember about the respiratory system is that the “goal” is to exchange as much gas as possible.
To accomplish this, the surface itself (be it the gills of fish, the skin of frogs, of the lungs of mammals) must be thin,
moist, well-supplied with blood, and of substantial surface area. Keep these criteria in mind as you perform the
following exercises.
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EXERCISE 7: Respiratory Structures In Mammals (1.5 points)
In this exercise you will examine the respiratory structures of a representative mammal, either a cat or a
fetal pig. A model of the human respiratory system will also be on display for you to make comparisons.
Materials:
Cat or fetal pig
Blunt probe
Gloves
Prepared slide of human bronchioles
Procedure:
1. Obtain a preserved fetal pig from your instructor. If the pig is not yet dissected, have your instructor
explain the correct process for the initial dissection, the neck region should be carefully dissected to
expose the respiratory structures.
2. Examine the oral and nasal cavities. Note that they are connected in back, at the PHARYNX, and that
the nasal cavity is partitioned internally to increase surface area.
3. Locate the LARYNX; notice the location of the esophagus and the trachea relative to the pharynx.
Make a cut along one side of the larynx to observe the complex arrangement of muscles and cartilage
responsible for proper function. (You can get an idea of the complexity of movement of this structure
by holding your finger gently against various portions of your larynx as you swallow).
4. Examine the TRACHEA. Note the numerous cartilage rings supporting the walls of the trachea,
squeeze both the trachea and the esophagus. To get an idea of the effectiveness of these rings in
keeping the “windpipe” open (you can feel these rings along the portion of your trachea in the cleft just
above your rib cage and below your larynx).
5. Observe the external structure of the LUNGS; they are composed of several lobes, each essentially
identical in structure and function (but not necessarily in appearance). Note that the lungs are nestled
within the thoracic cavity, immediately adjacent to the heart; examine the connections between the
lung and heart closely to locate the pulmonary blood vessels. Note the muscles between the ribs
(intercostal), which are partially responsible for ventilation of the lungs. The lungs are separated from
the digestive organs by a rather thin sheet of muscles, the DIAPHRAGM, which is also responsible
for ventilation. Examine the diaphragm.
6. Follow the trachea downward, into the lungs. Here they split into a pair of BRONCHI, which soon
split again into smaller and smaller BRONCHIOLES. Make several cross sections of one lobe to
observe the branching nature of these tubes; you will also note the latex injected into the pulmonary
arteries and veins.
7. View the prepared slide of human bronchioles to gain an appreciation of the internal structure.
Questions:
1. List the functions of the following:
Nasal cavity:
Pharynx:
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Larynx:
Trachea:
Bronchi:
2. Why didn’t you observe the alveoli of the lungs?
3. Why does the trachea have cartilage rings, but the esophagus doesn’t?
4. How is air inhaled and exhaled by your lungs? What organ systems are involved? How do they
contribute to inhalation and exhalation?
5. Describe the pulmonary circuit. Which blood vessels are involved?
THE DIGESTIVE SYSTEM
Digestion is the process by which food substances are broken down into forms which can be readily
absorbed through cell membranes into the body, the digestive system is the suite of organs which accomplish this
task. The digestive system is composed of a rather long muscular tube, the ALIMENTARY CANAL, which
extends from the mouth to the anus; both the name and the functions of the tube change along its length. The
portions of the canal we will observe include the mouth, pharynx, ESOPHAGUS, STOMACH, SMALL
INTESTINE, and LARGE INTESTINE. A number of ACCESSORY ORGANS, which secrete materials into the
alimentary canal to aid in transport and digestion, are attached along its length. The accessory organs you will
observe include the LIVER, GALL BLADDER, and PANCREAS.
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EXERCISE 8: Digestive Structures In Mammals (1.5 points)
In this exercise you will observe the major organs of the digestive system. Since the digestive system
functions much like an assembly line, it is most effective to begin at the mouth and progress toward the anus, and to
review the function of each of the organs as they are encountered (use your text book to review the function of the
organs).
Materials:
Cat or fetal pig
Blunt probe
Gloves
Preserved slide of small intestine
Procedure:
1. Examine the oral cavity, including the tongue. You may wish to dissect away part of one side of the
face to facilitate viewing. Cut through the pharynx as far down as the larynx, where the esophagus
joins the trachea.
2. Follow the esophagus down from the larynx; it lies deep within the body, partially hidden by the heart
and lungs. It soon disappears into the diaphragm; if you look on the opposite side of the diaphragm,
however, you will notice the esophagus extending out to the stomach. Squeeze the esophagus at the
junction of the stomach; note the hardened cardiac sphincter, a circular bond of muscle which prevents
food from flowing back out of the stomach into the esophagus.
3. Note the size and shape of the stomach. Make an incision along the side of the stomach and look at the
internal lining. Squeeze the other end of the stomach, and note the presence of a second sphincter, the
pyloric sphincter. (You may make a cut through the pyloric sphincter to see the nature of the muscle –
begin the cut in the stomach and end it in the intestine).
4. Just beyond the stomach is the small intestine, named for its diameter and not its length (it is much
longer yet thinner than the large intestine). Near the junction of the stomach and small intestine, two
accessory organs secrete materials into the intestine by way of rather hard-to-find ducts (thin tubes).
Locate the pancreas within the membrane along the margin of the small intestine and note the texture
of the internal lining.
5. Follow the small intestine down, through numerous turns and twists, to the point where it meets the
large intestine. You may gently pull the intestines out if their membranous sheath if necessary. Follow
the large intestine to the end of the animal; this bit if the alimentary canal is the rectum, which
eliminates the feces out through the anus.
6. Note the presence of blood vessels serving all of the organs listed above.
7. View the prepared slide of small intestine to gain an appreciation of the large surface area of this
structure.
Questions:
1. What are the functions of the organs of the digestive system (alimentary canal) indicated below?
Alimentary canal:
Esophagus:
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Stomach:
Small intestine:
Large Intestine:
2. State the accessory organs and indicate the function(s) of each one?
3. How is food moved along in the alimentary canal (what type of muscle contributes)?
4. Which organs in the digestive system seem to have the largest blood supply? Why do you think this is
so?
URINARY SYSTEM
The urinary system is responsible for filtering all of your blood many times daily, returning the “good”
materials to the general circulation while removing the wastes (salts, urea, etc.) in a concentrated solution called
urine. The urinary organs in your body responsible for the filtering and excretion are the KIDNEYS, while the
URETERS, URINARY BLADDER, and URETHRA are responsible for transporting and storing the urine prior to
elimination. We will take a brief look at these structures in a representative mammal.
EXERCISE 9: Urinary Structures In Mammals (1.5 points)

Materials:
Cat or fetal pig
Blunt probe
Gloves
Procedure:
1. Locate the kidneys along the back wall of the abdominal cavity.
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2. Gently (with your fingers) pull the kidney away from the body wall, pulling the connective tissue away
as the kidney is pulled free. Note the presence of at least two major blood vessels, the renal arteries and
renal veins.
3. As you lift the kidneys, note a long thin tube running down toward the urinary bladder (this tube may
be encased in a membrane). This tube is called the ureter. A single tube, the much larger urethra, runs
from the bladder out through the body wall to the external urinary orifice.
Questions:
1. What are the functions of the organs of the urinary system indicated below?
Kidney:
Ureters:
Urethra:
Bladder:
2. Where might you expect to find sphincter muscles in this system?
3. Why were the blood vessels serving the kidneys so large (relative to those serving most of the other
organs you have viewed in this lab)?
4. Out of the four systems studied today (circulatory, respiratory, digestive and urinary), which could be
shut down for the longest period of time without causing harm to the body? Which is the shortest
period of time? Why?
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Animal Anatomy
Systems involved in the exchange of materials between cells and the environment:
1) Circulatory system- moves blood around the body
a) Blood - circulating fluid
i) RBCs deliver oxygen to the body and remove carbon dioxide from the body
ii) WBCs - part of immune system
b) Vessels to move fluid
i) Arteries - move blood away from the heart
ii) Veins - move blood to the heart
iii) Capillaries - connect arteries and veins
c) Heart- pump to keep fluid circulating
i) Atria - 2 upper chambers
ii) Ventricles - 2 lower chambers
iii) Right side pumps to the lungs (pulmonary circulation) to unload CO 2 and pick up O2
iv) Left side pumps to the body (systemic circulation) to deliver O 2
d) Blood Flow
Body→Inferior and Superior Vena Cava→Right Atrium→Right Ventricle→Pulmonary
Artery→Lungs→Pulmonary Vein→Left Atrium→Left Ventricle→Aorta→Body
2) Respiratory system - supplies O2 to the blood and eliminates CO2

a) Oral and nasal cavities
b) Larynx
c) Trachea
d) Bronchi; bronchioles
e) Lungs - contain alveoli - sacs in lungs; site of gas exchange
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3) Digestive system - breaks down food to be absorbed by body cells for nutrients
a) Alimentary canal - extends from mouth to anus
i) Mouth
ii) Pharynx
iii) Esophagus
iv) Stomach
v) Small intestine
vi) Large intestine
vii) Accessory organs - salivary glands, liver, gall bladder, pancreas
4) Urinary system - filters blood to remove wastes as urine

a) Kidneys - do the filtering
b) Transport and storage of urine
i) Ureters
ii) Urinary bladder
iii) Urethra
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PLANT ANATOMY
Everyone is familiar with plants, yet very few people really understand the plants around them. In the
current lab you will look at plants in great detail to gain a better understanding of how they are put together and how
they function. Plants, like animals, are composed of discrete tissues and organs. The organs of a plant include roots,
stem, leaves, and flowers. These organs are specialized to perform specific functions necessary for the growth,
survival and reproduction of the plant. ROOTS are designed to anchor the plant in the soil, to absorb water and
minerals from the soil and make them available for the rest of the plant, and in some cases to store food reserves for
future use by the plant. STEMS are designed to transport water and dissolved materials (such as carbohydrates,
amino acids and minerals) from one part of the plant to another. LEAVES are the main sites of photosynthesis and
gas exchange, and FLOWERS are the reproductive structures of plants.
PLANT ROOTS
Plants display a wide variety of root types, both within a single species as well as between species.
Examples of various root types include the carrot, which has a rather large taproot (mainly used for storage of
nutrients, which is why humans eat them), and numerous finer lateral roots, the site of the majority of uptake of
water and nutrients from the surrounding soil. Quite different from the carrot are the roots of rye grass, which have a
dense collection of thin fibrous roots growing down into the soil from the stem; these roots store very few nutrients,
and function mainly to absorb water and nutrients. For both root types (and the diverse array of intermediate root
types) the main sites of water and nutrient absorption is through the very fine ROOT HAIRS, millions of billions of
root hairs associated with the roots of every plant provide an enormous surface area cross which water and nutrients
can be absorbed for the soil.
EXERCISE 1: Anatomy Of The Root Tip (2 points)

The root tip is the site of root growth and the main site of water and nutrient absorption. The tip can be
divided into several regions based on the appearance and function of the cells. At the apex of the root is the ROOT
CAP. This is the portion of the root which is in contact with the soil that the root is “plowing through,” hence this
portion of the root is continually being worn away. The root cap serves to protect the underlying region, the
APICAL MERISTEM, which is responsible for producing all of the new cells which make up the root as it grows
(including the constant production of new root cap cells to replace those worn away during growth). The apical
meristem is located within a region known as the ZONE OF CELL DIVISION; as the name implies, this is the
region within the root where the majority of mitotic cell division is occurring, resulting in an increase in cell
number. Beyond this region (i.e., in the region where cells are just a bit older) is the ZONE OF ELONGATION,
where the newly formed cells increase in size quite dramatically; it is this increase in size which “pushes” the tip of
the growing root deeper and deeper into the ground. Beyond this region is a ZONE OF MATURATION, where the
cells specialize and take on their specific functions (epidermis, xylem, phloem, etc.). In the following exercise you
will observe the portions of the root tip on a prepared slide of an onion tip, and compare what you see in the
microscope to some examples of living roots.
Materials:
Prepared slide of onion root tip
Radish seedling with root exposed
Microscope slide
Cover slip
Razor blade
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Procedure:
1. Obtain a prepared slide of the onion root tip.
2. Beginning at the apex (pointed end) of the root, locate and draw the following regions: root cap, apical
meristem, zone of cell division, zone of elongation, and zone of maturation.
3. Search along the upper end of the root to find extensions of the epidermal cells, the root hairs. Draw
them on the root above.
4. Obtain a growing radish seedling. Examine the very tip of the root with your bare eyes. Draw what you
see.
5. Make a wet mount of the terminal ½ inch of the root tip, being very careful not to damage the fine root
hair. Observe under magnification and draw below.
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Questions:
1. Why do the cells look different in the various regions of the root?
2. What would happen of a root could not produce a root cap?
3. What region of the root tip is actively involved with absorbing and transporting water?
4. How many root hairs can you see on the living root tip?
EXERCISE 2: Monocot And Dicot Root Anatomy (2 points)
MONOCOTS and DICOTS are two major groups of plants which differ in root, leaf ,and flower
morphology. In the current lab we will examine cross sections of mature roots of both monocots and dicots; this
means the roots you will view have been cut in the zone of maturation, after al of the cells have differentiated and
are performing their specialized functions.
Although the arrangement of tissues within the roots differs between monocots and dicots, the factions are
basically the same. XYLEM is a vascular tissue responsible for transporting water (and dissolved nutrients) from
the site of absorption up to leaves. PHLOEM is a vascular tissue which carries water and dissolved materials
(carbohydrates, amino acids, etc.) in either direction – from the roots to the leaves or from the leaves to the roots.
There are two major barriers to the flow of water and nutrients at the level of the root: the external EPIDERMIS
(including the extensions of epidermal cells, or root hairs, in smaller roots), and the inner ENDODERMIS (literally
“inside skin”), both of which function to regulate the movement of dissolved materials into the interior portion of the
root, where the xylem and phloem are located. The remainder of the cells in the root are largely PARENCHYMA
cells, which are responsible for storage of nutrients.
Materials:
Prepared slide of monocot and dicot root
Carrot
Razor blade
Procedure:
1. Observe the prepared slide of the monocot and dicot root. Locate, draw and label the xylem, phloem,
epidermis, endodermis, and parenchyma.
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Monocot root Dicot root
Questions:
1. How does the monocot root differ from the dicot root internally?
2. Why is the xylem inside the endodermis?
3. Using your drawings of the root tip (Exercise 1) and the roots cross sections, follow the path of flow of
water from the soil surrounding the root, into the vascular tissue, and up through the root on its way to
the leaves.
4. What plants are you familiar with which store energy in their roots? Do they do it for humans?
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PLANT STEMS
Stems are the organs which help the plant remain upright, thus providing maximum exposure of the leaves
to the sun. In a way, stems are like upside-down roots; they grow from the tip (at an apical meristem), contain xylem
and phloem to transport materials, and are surrounded by an epidermis. Likewise, monocot and dicot stems differ
more in appearance than in function, except that monocots do not tend to develop secondary growth (“wood”). In
the current exercise you will examine the structure of both monocot and dicot stems. Keep in mind throughout that
the structures you are looking at (like xylem and phloem) are continuous with those you just looked at in the roots:
they form set of continuous, unbranched tubes from the root tips to the leaves.
EXERCISE 3: Monocot And Dicot Stem Anatomy (2 points)

Monocot and dicot stems, like roots, have different arrangements of xylem and phloem. In monocots,
bundles of both xylem and phloem (which look like “monkey faces”) are distributed throughout the interior of the
stem. In contrast, dicots are arranged so that the xylem and sloughing off the “old” phloem yearly with the bark.
This exercise will focus on the primary (non-woody) growth, but you should have a feel for the nature of the
secondary (woody) growth seen in the trees and “wood” all around you.
Materials:
Prepared slide of a monocot and dicot stem
Prepared slide of a young dicot stem
Prepared slide of a woody dicot stem
Blocks of wood
Microscope slide
Cover slip
Razor blade
Celery
Procedure:
1. Examine the prepared slide of the monocot and dicot stems. Locate, draw and label the epidermis,
xylem, phloem, and parenchyma.
Monocot stem Dicot stem
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2. Examine the prepared slide of the woody dicot stem. Identify, draw and label the yearly growth rings
of xylem tissue and the phloem.
3. Examine several blocks of wood with your bare eyes. With the information gained from microscopic
examinations of cross sections of wood, identify the various structures you see (rings, bark, etc.) make
simple labeled sketches.
4. Obtain a piece of celery which has been submerged in water containing food coloring. Make several
cross-sections along the stem (all the way up to the leaves), and make some cuts at angles to the stem.
Draw and label what you see.
Questions:
1. How do non-woody dicot and monocot stems differ?
2. Try to explain what the pattern of xylem and phloem “tubes” would look like if you could make a
series of cross sections between the roots and the stems of monocots and dicots.
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3. Using information you have learned in this exercise, try to explain the pattern(s) of grain on various
pieces of wood around the classroom (which way is “up and down”? at what angle was the wood cut?
What are the “lines”? etc.).
4. How did the celery compare to the prepared slides? Does the stem look like that of a monocot or dicot?
EXERCISE 4: Dicot Stem Tip Anatomy (2 points)
Like roots, stems grow in length from the tips. The SHHOT APICAL MERISTEM, located at the tip of
each stem, is where the majority of cell division, elongation, and maturation occur. In contrast to roots, one of the
maturation products in stem tips is that of leaves, in the form of LEAF PRIMORDIA; in the angle of each leaf
primordia, a small bit of meristematic tissue (the BUD PRIMORDIUM) is left behind and often remains dormant
until the original meristem is removed or dies, at which time the bud primordium becomes a new shoot apical
meristem and produces a lateral branch.
Materials:
Prepared slide of Coleus stem tip
Samples of stem tips from living plants
Procedure:
1. Examine the prepared slide of the shoot tip. Locate and draw the apical meristem, leaf primordia,
vascular tissue, and bud primordium.
2. Obtain examples of living tips available at the front of the room. With your bare eyes, look for the
external structures identified in Step 1 above.
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Questions:
1. As you move away from the tip of the stem, why do the cells change in appearance? What structures
would you except to find if you could follow the stem down further?
2. Look at the growing tips of the plants at the front of the lab. Note the appearance of the tip and any bud
primordia. How do the differences in the shoot tips relate to differences in leaf number and
arrangement characteristic of each species?
PLANT LEAVES
The leaf is an organ specialized to carry out photosynthesis. Since photosynthesis requires light, and light
does not penetrate very far into living tissues, most leaves are relativity thin; in addition, the CHLOROPLASTS
(the actual site of photosynthesis) are concentrated in cells along the uppermost layer of most leaves. Photosynthesis
also requires carbon dioxide and water; water is supplied through the vascular tissues we have been looking at, but
the carbon dioxide is gained directly from the air by the leaves through pores called STOMATA.
The relatively large surface area of a leaf, combined with the presence of numerous tiny holes in the
epidermis, produces a potentially serious problem – water loss by evaporation. To minimize evaporative loss, plants
typically have two adaptations: 1) a hydrophobic, non-cellular, waterproof layer called the CUTICLE (secreted by
the epidermis) surrounding the entire leaf, and 2) a pair of expandable, bean-shaped GUARD CELLS surrounding
the stomata which open and close to regulate the movement of gases in and out of the leaf. In this exercise, you will
examine preserved and living leaves with an eye toward understanding both the function and structure of leaves.
EXERCISE 5: Leaf Structure (2 points)

A typical leaf is surrounded by a thin (one cell thick) epidermis; the epidermis may itself be covered with
fine hairs and/or a waxy cuticle to minimize water loss. The area between the upper and lower epidermal surfaces is
termed the mesophyll (literally “middle leaf”), which is arranged into two discrete layers. The upper layer, the
PALISADE PARENCHYMA, is located just beneath the epidermis on the top surface of the leaf, and is composed
of a single layer of tightly packed, column-shaped cells. It is within this layer that the majority of photosynthesis
occurs. The lower layer of cells, called the SPONGY PARENCHUYMA, is composed or rounder, loosely packed
cells. The air space around each cell maximizes the surface area available for gas exchange and allows air to move
directly to the palisade cells.
Guard cells are modified epidermal cells, and are found mainly along the bottom surface of leaves. The
guard cells swell and shrink in response to the water and carbon dioxide needs of the plant. When the guard cells are
swollen, the stomata are open and carbon dioxide enters and water leaves the leaf; when the guard cells are flaccid,
stomata are closed, thus preventing the movement of both water and carbon dioxide into or out of the leaf. By
regulating the number and duration of stomatal openings, a plant is able to effectively control water loss.
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Of course, the water which is being lost from the leaf is that which was absorbed at the root hairs and
transported to the leaves in the xylem. Likewise, the photosynthetic products (mainly carbohydrates) produced in the
leaves are transported from the leaf in the phloem. Thus, the leaf must be well supplied with vascular tissue.
Numerous vascular bundles (bundles of xylem and phloem) enter and radiate out through the leaf.
Materials:
Prepared slide of leaf
Microscope slide
Cover slip
Razor blade
Forceps
Leaves from a geranium and other plants
Procedure:
1. Examine the prepared slide of a leaf cross section. Locate, draw and label the epidermis, palisade
parenchyma, spongy parenchyma, guard cells, stomata, chloroplasts, and vascular bundle.
Questions:
1. Why are the parenchyma cells so tightly packed along the top surface of the leaf, yet space so loosely
in the lower spongy layer? Which had the most chloroplasts?
2. How many vascular bundles did you see in the leaf, and were they all the same size?
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3. Why do you think most of the guard cells in the preserved leaf were closed? How did this compare to
the living leaf you viewed?
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Plant Anatomy
Plant Organs include Roots, Stems, Leaves, and Flowers

1) Roots - anchorage, absorption of water and minerals from soil, food storage
a) Root types
i) Taproots- storage of nutrients
ii) Lateral roots- uptake of water and nutrients
iii) Fibrous roots- grow downward to absorb water and nutrients
iv) Root hairs- structure that allows for water and nutrient uptake by increasing surface
area; found at the root tip
b) Root cap- found at apex of root; protects the root (specifically, the apical meristem) as the
root plows through the soil
c) Zone of cell division- contains apical meristem that produces all new cells of the root
d) Zone of elongation- cells get increasing larger in this region to push the tip of the root deeper
into the soil
e) Zone of maturation- cells become specialized (become xylem, phloem, epidermis, etc)
f) Tissues found within the root

i) Monocots and dicots differ in the arrangement of differentiated tissue
ii) Xylem- vascular tissue that transports water and dissolved nutrients to the leaves
iii) Phloem- vascular tissue that carries nutrients
iv) Epidermis and endodermis regulate the movement of dissolved materials into the
interior portion of the root (xylem and phloem)
v) Parenchyma cells- the remainder of the root which stores nutrients
4X 40X
Cross section - dicot root
Cross section - monocot root
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2) Stems (aka shoots) - transport water and dissolved materials (carbs, minerals, amino acids)
a) Comparative anatomy of monocots and dicots
i) Monocot stems have bundles of xylem and phloem distributed throughout the
interior
ii) Dicot stems have xylem on the interior portion of the stem and phloem around the
periphery. They are also capable of secondary growth (wood)
4X 40X
4X 40X
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3) Leaves- photosynthesis and gas exchange

a) Chloroplasts - concentrated in cells along the uppermost layer of leaves
b) Stomata - pores that allow carbon essential gases into and out of the leaves
c) Guard cells - modified epidermal cells that surround stomata to open and close the pore in
response to water and CO2 needs to reduce evaporation and regulate gas exchange; found
mainly on the bottom surface of leaves
d) Cuticle - waterproof layer surround entire leaf to reduce evaporation
e) Epidermis - outer layer of leaf; one cell thick; may be covered in fine hairs and/or waxy
cuticle
f) Mesophyll (means middle leaf) - area between upper and lower epidermis; formed by two
layers
i) Palisade parenchyma - upper layer; single layer of tighly packed column-shaped
cells; most photosynthesis occurs here
ii) Spongy parenchyma - lower layer; rounder, loosely packed cells; air spaces provide
surface area for gas exchange and allow movement of air to palisade cells
g) Vascular bundles - xylem and phloem bring water to the leaf and take carbohydrates out of
the leaf
4) Flowers - reproductive structures

a) They are pretty, but we are not looking at them in lab!
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METABOLISM
METABOLISM is the sum total of all the chemical reactions which happen within a cell, including those
reactions which build molecules and cells and those reactions which break down molecules and cells. ANABOLIC
METABOLISM, or anabolism, involves the “building” processes, while CATABOLIC METABOLISM, or
catabolism, involves breakdown processes. We will look at examples of both of these metabolic processes in the
current lab.
There is a very large number and a wide variety of anabolic and catabolic reactions which are necessary to
keep organisms alive. Some of the more important include digestion (the breakdown of food into molecules small
enough to be absorbed into the blood), the building of new cells for growth and reproduction, photosynthesis (the
production of carbohydrates by plants) summarized by the following equation:
6 CO2 + 6 H2O → 6 C6H12O6 + 6 O2

(carbon dioxide) (water) (sugar) (oxygen)
and respiration (the breakdown of carbohydrates within cells to provide energy in the form of ATP for cellular
work) summarized by the following equation:
6 O2 + C6H12O6 + 36 ADP → 6 CO2 + 6 H2O + 36 ATP

(oxygen) (sugar) (carbon dioxide) (water) (energy)
(in the case of respiration shown above, catabolism of sugar and anabolism of ATP are coupled reactions, with the
energy lost from the sugars being trapped by the chemical bonds of ATP).
Plants carry out both respiration and photosynthesis, while animals only respire. Thus, plants produce
oxygen and carbohydrates (in photosynthesis) and use oxygen and carbohydrates (in respiration); for a plant to
grow, net photosynthesis (anabolism) must exceed net respiration (catabolism). Since animals cannot produce
carbohydrates or oxygen yet must consume both, they must obtain them from the surrounding environment. Thus
animals need to eat and breathe. In animals, like plants, if the net anabolism exceeds the net catabolism, then the
animal grows (much like a bank account, with deposits and withdrawals representing anabolism and catabolism,
respectively).
Metabolic activities are carried out from us by ENZYMES. Enzymes are special protein molecules, called
CATALYTS, which help chemical reactions proceed. Without enzymes the reactions would happen at such a slow
rate that the cells (hence organisms) could not gain enough energy to stay alive for long. Although enzymes
participate in chemical reactions (i.e., they are the “builders” and the “breakers” of molecules and cells), the
enzymes themselves are unchanged by the reaction. A single enzyme molecule can catalyze thousands or millions of
reactions without being “used up”.
The rates of enzyme activity (hence metabolic activity) are affected by several environmental factors, such
as temperature, acidity (pH), and salinity. Most enzymes work well over only a narrow range of these and other
environmental factors, and may be completely disabled under extreme conditions. This is one of the reasons your
body’s composition and temperature are maintained so precisely, for if your metabolic enzymes fail, you do too!
Most experiments dealing with anabolism and catabolism are a bit too elaborate or time consuming for
Biology 100 labs, however, in the following exercises we will conduct some basic experiments on plant metabolism,
as well as examine the role of enzymes in metabolic function.
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PHOTSYNTHESIS
As stated earlier, photosynthesis involves the capture of energy in the form of sunlight. This energy is
captured by special molecules, called PIGMENTS, which are very efficient at absorbing light of a particular
wavelength (i.e., color). Contained energy trapped by the pigments is transferred to the bonds in carbohydrates
which are the source of energy for all organisms on earth!
Although the process of photosynthesis looks like a single step,
6 CO2 + 6 H2O + sunlight → 6 C6H12O6 + 6 O2
it is really an extremely complex series of reactions. These reactions can be divided into two major groups: 1) those
requiring light to trap the energy and get the process going, and 2) those not requiring light but involved with
building the carbohydrate, using the trapped energy. These are conveniently referred to as the “Light” (light
dependent) and “Dark”(light-independent) reactions. As the simplified diagram below shows, the links between
Light and Dark reactions are “energy carriers”; the energy from sunlight is transferred to NADP+ and ADP (forming
NADPH and ATP, which are much higher in energy than ADP and NADP). These energy carriers perform as their
name implies – they just “pick up” energy in one part of the chloroplast and carry it to another part to be used for
anabolism of carbohydrates from carbon dioxide.
ATP* C6H12O6
O2 NADPH
*
Dark
Light Reactions
Sunlight*
Reactions
ADP
H2O NADP
CO2
* = High in energy
The product of photosynthesis is glucose, a carbohydrate. Glucose can be 1) converted into starch for
storage, 2) converted into cellulose for cell walls, 3) converted into proteins and lipids for the cell’s other
biochemical needs, or 4) “burned” as a fuel in aerobic respiration.
EXERCISE 1: Chloroplast: Site of Photosynthesis (1.5 points)
Chloroplasts are specialized organelles in most plant cells that are involved in photosynthesis. They contain
light-sensitive chlorophyll pigments that absorb rays of light. According to the direction and the angle of the light
rays, chloroplasts change their positions in a photosynthetic cell in order to absorb optimum lighting more
efficiently. Chloroplasts change their positions with the aid of cytoskeleton system in the cell. Myosin filaments
attach chloroplasts with them. Flowing of chloroplasts to new positions in the cell is an example of
CYTOPLASMIC STREAMING.
In this exercise you will observe chloroplasts and cytoplasmic streaming. In addition you will observe other
cell structures such as the cell wall. The nucleus and the location of the central vacuole in Elodea leaf cells.
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Materials:
Microscope slide
Cover slip
Elodea leaf
Procedure:
1. Remove a leaf from Elodea plant and make a wet mount
2. Examine under the low power and then the high power objective.
3. Locate chloroplasts and notice if they change their positions inside the cell (cytoplasmic streaming).
4. Indicate the cellular structures that you recognize. Describe cytoplasmic streaming in Elodea cells.
Explain your observations in the space below.
5. Draw and label the features of an Elodea leaf cell in the space below.
EXERCISE 2: Chlorophyll and Photosynthesis (1.5 points)

Chlorophyll is the primary photosynthesis pigment of photosynthesis. This pigment is stored in chloroplasts
and synthesis of starch cannot take place without it. In Exercise 1, you observed the chloroplasts in Elodea leaves. In
this exercise you will observe the relation between chlorophyll distribution and starch synthesis in Coleus leaves.
Materials:
Coleus leaves
Iodine (IKI)
250 ml beaker
95% ethanol
Syracuse culture dish
Timer
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Procedure:
1. Obtain a Coleus leaf.
2. Make a drawing of the leaf showing the distribution of chlorophyll in the space below.
3. Form a hypothesis regarding the distribution of starch in Coleus leaf. State your hypothesis in the
space below.
4. Boil the leaf in water for 3 minutes
5. Boil the leaf in 95% ethanol for 6 minutes.
6. Place the leaf in Syracuse culture dish and flood with potassium iodine solution for 3 minutes.
7. Make a drawing of the leaf showing the distribution of starch in the space below.
Questions:
1. What part(s) of the leaf tested positive for starch?
2. Did your results support your hypothesis? Explain.
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EXERCISE 3: Isolation of Photosynthetic Pigments (1.5 points)

Light energy, a form of electromagnetic radiation, must be converted to chemical bond energy before it is
usable by cells. However, before the energy can be converted, it must first be trapped. This is the role of
PHOTOSYNTHETIC PIGMENTS, which are contained in an organelle called the CHLORPLAST. The main
photosynthetic pigment is CHLOROPHYLL, which appears green to our eyes because it does not absorb (i.e., it
reflects) green light. However, there are a number of other pigments in most plant leaves as well, including
carotenoids (orange), anthocyanin (red) and xanthophylls (yellow). Normally these pigments are in low
concentration and distributed throughout the leaf, and there is usually much more chlorophyll , so the other colors
are masked. However, in the autumn, as leaves are beginning to fall, the chlorophylls are broken down, leaving the
reds and yellow behind in a spectacular display of leaf pigments.
In the current exercise you will separate the different pigments from spinach leaves by a process known as
PAPER CHROMATOGRAPHY. The idea is rather simple: mixtures of pigments are placed on a long strip of
paper and a column of solvent is then allowed to wick up through the paper. As the solvent passes the pigment
mixture, it will carry with it those pigments which are highly soluble in the particular solvent, and leave behind
those which are not soluble. The pigments we will isolate from spinach are all partially soluble in the solvent, but to
different degrees; thus, different pigments will move (“migrate”) different distances depending on their individual
solubilities, and the pigments will be separated, with the most soluble pigments moving further up the paper than the
least soluble pigments. When the experiment is complete, a number of colored bands should be visible on the
chromatography paper.
Materials:
Chlorophyll extract (from spinach)
Large test tube with stopper
Piece of chromatography paper
Small paintbrush
Metric ruler
Procedure:
1. Obtain a large test tube (with a stopper) and a piece of chromatography paper. Handle the paper by the
edges only (of with a pair of forceps), as oils from your finger may affect the results.
2. Cut the chromatography paper into a “V” at the bottom end (see the sample at the front of the room).
With a pencil, make a faint line across the paper about one inch from the pointed end. This is your
“origin”, the place where you will put the pigments.
3. Go to the front bench and get several drops of the chlorophyll extract and a paintbrush. Apply a
narrow strip of the extract to the paper directly on the pencil line. Allow the extract to dry (blowing
gently will speed this up), and reapply 5or 6 times until the line is rather dark with pigments. DO NOT
apply so much in any one application that the line spreads too much; try to keep the line as narrow as
possible for the best results.
4. Hook the top of the paper strip to the paper clip suspended from the stopper. Place the stopper tightly
on the test tube, allowing the paper to hang down within the tube (the paper should not quite reach the
bottom of the tube - - if it does, trim the strip form the top). Mark on the outside of your tube (with a
wax pencil) where the bottom tip of the strip reaches. Remove the stopper and all solvent to about ½
inch ABOVE the line you just marked, then replace the stopper and strip. The solvent should NOT be
high enough to reach the pigments!
NOTE: THE SOLVENTS ARE TOXIC, SO MINIMIZE EXPOSURE TO THEM BY KEEPING

YOUR TUBES CAPPED AT ALL TIMES WHEN THEY HAVE SOLVENTS IN THEM, AND
DISPOSE OF SOLVENTS IN THE APPROPRIATE CONTAINER WHEN FINISHED.
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5. Place the test tube in a rack near the window, being very careful not to shake or tip the tube when
handling. Observe your tube every 5 or 10 minutes, noting the movement of the solvent “front” up the
paper strip. When the front is near the top of the strip, remove the strip from the tube make a small
pencil mark at the solvent front (the solvent will disappear rather fast by evaporation), and observe the
pigments under both room lighting and ultraviolet light.
6. Make a drawing of what you observe, noting the distances between the origin, the pigments and the
solvent front.
EXERCISE 4: Starch Production In Leaves (1.5 points)

This exercise deals with the dark (light-independent) reactions, where the energy trapped from sunlight (by
pigments in the light reactions) is actually used to “stick” carbon atoms together to form the high-energy molecules
of glucose. Without the energy from the sun, the dark reactions could not proceed. Remember that plants need to
“burn” this glucose to produce ATP and they store extra glucose as starch. Therefore, if plants do not receive
sufficient amounts of light, they are not able to produce glucose, and eventually will use up their stored glucose
(starch) and die. In the following experiment, portions of plant leaves were covered for a period of time, and the
plants were exposed to normal periods of sunlight. You will examine the covered and uncovered portions of the leaf
to see if there are differences in the amount of starch within the leaf.
To determine the presence of starch, we will stain the leaves with iodine, which turns deep blue-black in
the presence of starch but remains reddish in the absence of starch. However, because the green pigments will mask
the effects of iodine on starch, we will first clear the leaf of pigments and then apply iodine.
Materials:
Geranium leaf, partially covered
Iodine (IKI)
250 ml beakers
95% ethanol
Petri dish
Paper towels
Forceps
Procedure:
1. Obtain a partially covered geranium leaf from the plant at the side of the room.
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2. Make a scale drawing of the leaf, making the area covered by the paper.
3. Remove the covering and place the geranium leaf into a beaker of boiling water for 5 minutes.
4. Remove the leaf from the boiling water and place into 50 ml of hot (NOT BOILING!) ethanol for
several minutes.
5. Remove the leaf from the alcohol and place it on a paper towel to remove excess liquid. Place the leaf
into a Petri dish and pour several ml of iodine over the leaf.
6. Make a drawing of the geranium leaf now, marking the areas which turned dark blue and those which
did not.
Questions:
1. How do your two drawings (before and after) compare? Why?
2. Based on your results, does it seem like leaves are able to move starches from one area to another
within a single leaf?
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STARCH DIGESTION
Digestion is the breakdown of large food particles, such as what you had for breakfast or lunch today, into
much smaller units. In order for the food you eat to get to the cells in your body, where the materials are ultimately
“burned” in respiration, you must first reduce it to a size small enough to cross the lining of your digestive tract and
move into the bloodstream. You can not have large chunks of pancakes or hamburger floating around in your blood!
Thus, the function of your digestive system (mainly the small intestine) is to reduce proteins to their individual
amino acids, complex carbohydrates to their component simples sugars, and triglycerides to fatty acids which are
small enough to move into the bloodstream and to the cells where they are needed. The process of digestion, like all
other metabolic processes, is carried out by enzymes – digestive enzymes in this case. Digestive enzymes are
produced by salivary glands in the mouth, the lining of the stomach and small intestine, and the pancreas. They are
secreted into the digestive tract in response to the presence of food. Like other enzymes, digestive enzymes only
work within relatively narrow ranges of temperature and pH.
EXERCISE 1: Starch Digestion By Human Amylase (4 points)

In this exercise, you will examine the process of starch digestion by enzymes present in saliva. The enzyme
SALIVARY AMYLASE is responsible for breaking down of starch into simple sugars. You will donate amylase
yourself, and will test its effectiveness in breaking down a starch solution. We can test solutions for the presence of
starch (as in the previous exercise, using iodine) and the presence of sugars, using a solution called BENEDICT’S
SOLUTION. Benedict’s solution is a blue solution that, after boiling, turns orange or brick red in the presence of
sugar. You will also examine the effects of temperature and pH on amylase activity.
Materials:
8 Test tubes
Test tube rack
250 ml beaker of water
Funnel
Metric ruler
Pipette
HCL (hydrochloric acid)
Starch solution
Benedict’s solution
Iodine (IKI)
Wax pencil
Saliva (from a member of your group)
Procedure:
1. Label the test tubes 1 through 8.
2. With the wax pencil and ruler, mark each tube at the 1cm and 2 cm mark from the bottom (see the
example in front).
3. Have one (or more) volunteer(s) spit into a small beaker; you will need about 15 ml of saliva.
4. Place 1 ml of amylase (spit) into tubes 3, 4, 5, 6, 7, and 8.
5. Place 1 ml of water into tubes 1 and 2. Mix by swirling, and set them aside in the rack.
6. Place tubes 5 and 6 in a boiling water bath for 5 minutes. Remove the tubes and place them in the rack.
7. Place ten drops of HCl in tubes 7 and 8. Mix by swirling. Set them aside in the rack.
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8. Place 1 ml of starch in tubes 1 through 8. Mix by swirling.
9. Place the rack (containing all 8 tubes) in a 37°C incubator for approximately 30 minutes.
10. Remove the tubes from the incubator and test for the presence of starch and sugars according to the
following directions:
STARCH: Add several drops of IKI to tubes 1, 3, 5 and 7. Mix thoroughly by swirling. Note
the colors of the tubes.
Tube Number Color of Tube Starch Present?

1
SUGAR: Add equal volumes of Benedict’s solution (about 2 or 3 ml) to tubes 2, 4, 6 and 8,
and mix by swirling. Place the tubes into a boiling water bath for several minutes. Keep an
eye on the solutions, and note any change in color.
Tube Number Color of Tube Sugar Present?

2
11. STARCH DIGESTION CLEANUP
a. POUR LIQUIDS FROM TUBES CONTAINING IKI INTO THE IKI WASTE JAR.
b. POUR LIQUIDS FROM TUBES CONTAINING BENEDICT’S SOLUTION INTO THE BENEDICT’S
WASTE JAR.
c. RINSE ALL TUBES THOROUGHLY AND PLACE UPSIDE DOWN IN THE TUBE DRYING RACK.
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Questions:
1. Which tubes showed the presence of sugar? Starch? Both sugar and starch?
2. Why did we boil tubes 3 and 4? What was the effect?
3. Why did we add HCl to tubes 5 and 6? What was the effect?
4. What was the reason for tubes 7 and 8?
5. Which tubes showed the highest rates of starch digestion?
6. Which tubes showed the lowest rates of starch digestion?
105
OSMOSIS EXPERIMENT
Both osmosis and diffusion are important biological processes because they are involved in the transport of
materials (such as oxygen, carbon dioxide) throughout your body, and also the transport of materials into and out of
individual cells. Osmosis and diffusion are related processes because both involve the moving of materials down a
CONCENTRATION GRADIENT (i.e., from areas of high concentration to areas of low concentration).
In osmosis the concentration gradient of water (the SOLVENT) is created by the addition of a dissolved
solid (the SOLUTE; e.g., salt or sugar) in two different solutions, separated by a selectively-permeable membrane
(like the plasma membrane). The more solute present in a solution, the less room for water, and thus the lower the
concentration of water. Conversely, a solution with lower concentrations of solutes will have a higher concentration
of water. In osmosis the solutes are prevented from diffusing by a selectively-permeable membrane, so only the
water diffuses. Water diffuses across a selectively-permeable membrane from the solution with lower solute
concentration into the solution with higher solute concentration.
The independent variables we will be testing in this lab are sucrose (solute) solutions of different
concentrations, each contained in a selectively-permeable, cellophane membrane. The dependent variable is the rate
of osmosis. The question asked is how different concentrations of sucrose (the independent variable) will affect the
rate of osmosis (the dependent variable). Our prediction is that as the sucrose concentration within the selectively-
permeable membranes increases, the rate of osmosis will also increase. In your group formulate a hypothesis based
on this prediction.
Materials:
0.25 M sucrose solution

Deionized water
4 beakers
4 pieces of dialysis tubing
8 pieces of string
Electronic balance
Markers
Labeling tape
Paper towels
Procedure (work in groups of 4 students):
Preparation of the (selectively) semi-permeable bags
1. Fill 4 separate beakers 150 ml with de-ionized water (di-H2O). Label each (with labeling tape) to
indicate the different sugar concentrations (0 M, 0.25 M, 0.5 M, and 0.75 M).
2. Prepare 4 separate dialysis tubing (selectively-permeable) bags.
a. Obtain tubing from beaker of distilled water (used to soften the tubing).
b. Twist and tie one end tightly with string to form an open bag.
c. Fill each bag approximately half full each with a different sucrose solution. (Do this at the
designated area to limit spills.)
d. Twist and tie up the open ends of the bags tightly with string. (Remember that the bags should
be able to swell so don’t tie the string right next to the sugar solution)
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e. Rinse off each bag with di-H2O to wash away any spilled sucrose solution.
Measurement of the rate of osmosis
Measurement of the rate of osmosis is done by evaluating the percent of the initial mass of each bag.
Percent of the initial mass of each bag is determined by:
1. Weighing each filled cellophane bag before you sink it in the appropriate beaker of di-H2O.
2. Weighing each bag after it has been left in the beaker of di-H2O for 35 minutes.
3. Dividing the final mass by the initial mass and multiplying by 100. (This procedure is repeated for
each of the four bags your group prepared.)
Final mass x 100 = % of the initial mass

Initial mass
4. Record your group’s data on the blackboard.
5. OSMOSIS EXPERIMENT CLEANUP
a. EMPTY THE CONTENTS OF THE DIALYSIS BAGS IN THE SINK AND THROW THE
EMPTY BAGS IN THE TRASH CAN.
b. REMOVE LABELING TAPE FROM BEAKERS, RINSE THEM IN THE SINK AND PLACE
THEM UPSIDE DOWN TO DRY
c. SPRAY AND WIPE DOWN ALL WORK AREAS WITH DISINFECTANT/CLEANER WHERE
SPILLS WERE POSSIBLE (BALANCES, LAB BENCHES, FLOOR, ETC).
d. REPORT ANY SPILLS ON THE FLOOR TO YOUR INSTRUCTOR IMMEDIATELY. THEY
WILL MAKE SURE THAT ALL SPILLS ARE CLEANED UP IMMEDIATELY TO PREVENT
INJURIES AND STICKY/DIRTY FLOORS.
RESULTS & WRITE-UP
Each student will prepare an individual report on this lab using the combined class data. This report should
follow the scientific report format as presented by your instructor and described in the report guidelines and
graphing data on pages 115-117 of the lab manual. To interpret your results you will be constructing a
simple trend-line graph.
The following are some suggested ideas to consider when writing up your reports.
 Summarize the effect of the sucrose concentration on the rate of osmosis using a trend-line.
 What unmeasured factors may have caused variation in the rate of osmosis even when the value of
the independent variable was the same? (Why were there differences in % of the initial mass
values for the same sugar concentrations?) Give examples. Use the class raw data to support your
opinion.
107
GROWTH OF BACTERIA EXPERIMENT

We share our environment with various and diverse microorganisms. Humans harbor various bacteria on
their skin, in their upper respiratory tract, and in their alimentary canals. There are very few, if any, environments in
nature in which bacteria do not exist. Bacteria have been isolated from such diverse environments ranging from
sulfur hot-springs associated with volcanic activity to the ultra cold waters of the Antarctic.
Before discussing the details of this lab it might be useful to review some terminology used in association
with doing science. In today’s lab you will be designing and performing an experiment to determine the effect of
antibacterial soap (a variable) on bacterial growth. The term variable describes the fact that these conditions may
change and upon changing may produce some noticeable effect. Some variables are independent variables. The
independent variable in our specific case is antibacterial soap which may influence bacterial growth (the dependent
variable). There are, as you can imagine, many of these independent variables, but in most experiments only a single
independent variable is considered at one time. Ideally all other independent variables not being studied in the
experiment should be kept constant, or controlled.
Let’s take a few minutes to discuss a specific question that you can ask about antibacterial soap related to
the bacterial growth. Use this question and propose a specific, testable, hypothesis (if...then). You will then test this
hypothesis in the following experiment.
TESTING YOUR HYPOTHESIS
This lab will involve discussion and set-up during the first week and the reading of the results on the
second week of the lab. You will be conducting an experiment to test your hypothesis in regards to the effectiveness
of hand-washing with antibacterial soap on the reduction of bacteria growth.
MATERIALS & METHODS
Week 1
The class will design this experiment including the following: the amount of antibacterial soap and the method
of hand washing.
Procedure:
Each student will prepare one plate as assigned.
1. Obtain an agar plate. Draw a line on the bottom of the plate dividing the plate into two equal halves.
Label the halves 1 and 2. Along the edge label the bottom of the plate with the following:
- students initials or names
- type of exposure
- date of exposure
- lab section day and time (e.g., Tue 12-3)
2. Lightly press your thumb onto the middle of the agar over the area marked 1. Don’t press too hard or
you will put your thumb into the agar (it has a jello-like consistency). Cover the plate right away. You
do not want air-born organisms to contaminate your results.
3. Wash your hands in the manner decided by the class. Air-dry your hands. Press your thumb onto the
middle of the agar over the area marked 2.
4. Cover the plate and turn it upside down and then place it on the tray provided for the class.
108
5. Let it sit at room temperature until the next lab period.
Week 2
The results for this experiment are gathered as a class.
1. Obtain your plate from the previous week and determine the density of the colonies and any diversity
you can see. Diversity will be determined by differences in colony color, shape, size and texture.
Compare the two halves.
2. Record your results on the board.
3. PLACE ALL USED PLATES IN THE BIO-HAZARDOUS WASTE CONTAINER. DO NOT THROW THEM IN THE
TRASH CAN!
RESULTS & REPORT
Each student will prepare an individual report on this lab using the combined class data. This report should
follow the scientific report format as presented by your instructor and described in the report guidelines and
graphing data on pages 114-115 of the lab manual.
109
ALLELOPATHY EXPERIMENT
(Chemical Warfare among the Plants)

Plants can be complex chemical factories. They may produce chemical scents and nutrients to attract
pollinators. They may produce and store poisons in their tissues to discourage potential browsers and parasites.
Plants also compete with other plants for sunlight, soil nutrients, and water, and since they do not have the option of
avoiding competition by running away (like animals) they resort to chemical warfare. To reduce competition, they
may produce chemicals that inhibit the germination and growth of neighboring plants. This last phenomenon is
termed ALLELOPATHY. This chemical warfare can be against other plants of the same species (intraspecific
competition) or plants of a different species (interspecific competition).
Plants may release allelopathic substances in a variety of ways, including 1) directly into the soil from their
roots, 2) by leaching from living or dead shoots into the surrounding soil, or 3) by passing volatile compounds into
the air that are then deposited on the soil or the surfaces of other plants when dew forms.
In this laboratory exercise, we will test for the presence of allelopathic chemicals in plant leaves and the
effect of these allelopathic chemicals (independent variable) on the germination and growth (dependent variable) of
radish seeds. Allelopathy appears to be best developed in perennial and woody species of more humid regions.
Materials:
Ziploc bags
Plant leaves
Electronic balance
Weigh boats
Distilled water
Blenders
Cheesecloth
Plastic funnels
125 ml plastic bottles
10 cm filter paper
Petri dishes
Radish seeds
100 ml graduated cylinders
10 ml graduated cylinders
Procedure:
Prior to lab
1. Collect leaves from one plant species. Pick fresh leaves (no stems) and fill up a sandwich-type plastic
bag. Bring the leaves to the lab.
Extraction of Allelopathic Compounds (work in groups of 2-3 students)
1. Weigh out 10 grams of leaves from one plant species and place the measured sample in a blender.
2. Add 100 ml of distilled water to the blender.
3. Hold the lid of the blender firmly on the blender and blend completely.
4. Place 2-3 squares folded cheesecloth into a funnel and set into a stock bottle.
110
5. Pour the blended mixture through the funnel/cheesecloth to filter the solution.
Testing of Allelopathic Effects
1. Place three pieces of filter paper in each bottom of two petri dishes.
2. In one petri dish (labeled with the name of the plant you collected), add 10 ml of the filtered solution
so that it soaks the filter paper layers.
3. In the second petri dish (labeled control), add 10 ml of distilled water so that the water soaks the filter
paper layers.
4. In each petri dish, place 20 radish seeds (Raphanus sativus) on top of the three pieces of filter paper so
that they are uniformly spaced.
5. Place another piece of filter paper on top of the seeds in both dishes.
6. Place the lids on the petri dishes to prevent excessive evaporation and store at room temperature. The
seeds will be given a one week germination period and then evaluated.
7. The lab tech will check the condition of the cultures periodically. If necessary, small amounts of
distilled water will be added to re-saturate the filter paper and prevent the cultures from drying out. If
fungus appears, cultures will be moved to a cooler incubation environment.
CLEANUP
1. RINSE THE BLENDER JAR, FUNNEL, STOCK BOTTLE, AND ANY OTHER ITEMS THAT HAVE PLANT
EXTRACT OVER THE STRAINER AND RETURN THEM TO THEIR DESIGNATED LOCATIONS.
RESULTS & REPORT
The results will be gathered after a 1 week growth period. Examine your cultures and fill in the requested
information on the lab data sheet. Each student will prepare an individual report on this lab using the data
from your group. This report should follow the scientific report format as presented by your instructor and
described in the report guidelines and graphing data on pages 114-115 of the lab manual.
111
ALLELOPATHY DATA SHEET Name __
Name of plant species tested: __
A. Experimental Group (treated with extract)
Total number of seeds tested
Number of seeds germinating
Percentage successful germination
Length of each seedling in mm
Average length of seedlings in mm
B. Control Group (treated with distilled water only)
Total number of seeds tested
Number of seeds germinating
Percentage successful germination
Length of each seedling in mm
Average length of seedlings in mm
112
VEGETATION ANALYSIS EXPERIMENT
Each species, through the process of natural selection and evolution, has arrived at a slightly different
solution for surviving in its habitat. In plants, for example, these solutions may involve differences in such details as
leaf shape and size, plant size and form, etc. There will also be similarities owing to the fact that each plant species
shares the same basic “plant plan”. They all photosynthesize and require the structures necessary for photosynthesis,
such as leaves, stems, etc.
In this lab we want to see if different environmental conditions affect the structure of a plant. Since leaves
are a plant’s primary structure for gathering sunlight for photosynthesis, we are going to investigate whether the
amount of sunlight has an effect on leaf structure. It may be predicted that in low sunlight conditions, the plants may
have larger leaves as a result of natural selection. The evolutionary logic behind this is that a plant with larger leaves
would be able to gather more sunlight, make more carbohydrates, and thus produce more flowers and seeds than a
plant with smaller leaves in the same habitat. This would increase that plant’s reproductive success and result in
larger leafed plants being more common in future generations in the low light density habitat.
For this lab we will be comparing two populations of Common Olive (Olea sp. leaves), each being exposed
to different amounts of sunlight. The Common Olive is native to Mediterranean regions. It is an evergreen that
produces edible fruit. This plant also grows successfully in habitats with Mediterranean climate, such as California.
Materials:
Common Olive (Olea sp.) leaves
Ziplock bag
Tape
Markers (Sharpies)
Compass
Electronic balances
Scissors
Paper
Procedure (work in groups of 4):
At the Field Site
Students will walk to the field site (north of campus across North Campus Circle) in order to collect leaf
samples. The lab instructor will lead the students to the field site. Each group will collect four leaf samples from
Common Olive (Olea sp.) plants. Two (2) leaves will be collected from plants exposed to low sun density
(north) and 2 leaves from plants exposed to high sun density (south).
The leaves will be collected as follows:
1. Using the compass, determine the north (low sunlight exposure) and south (high sunlight exposure)
sides of the olive grove.
2. Choose a Common Olive plant from the north side of the grove, locate the middle of the branch and
pick one leaf. Repeat this procedure on another Common Olive plant within the same habitat and place
the two leaves in the Ziploc bag and label it “North.”
3. Repeat step 2 on the south side of the olive grove. Label the bag “South.” Each group of students
should have four Common Olive leaves, two from each habitat.
113
In the Lab
We want to know if Common Olive leaves growing in low sunlight habitat have larger surface area compared
to the Common Olive leaves growing in high sunlight habitat.
1. Trace each leaf from the low sunlight habitat (North) onto paper. Cut around the outline and weigh
each paper leaf. Make sure to keep track
2. Cut out a square of 100 mm x 100 mm dimensions. Weigh the cut out square.
3. Calculate mass/surface area of the cut out square. This is your conversion factor.
Conversion factor (CF) = mass/surface area of the cut out square.
4. Now calculate Common Olive leaf surface area:
Common Olive leaf surface area = (mass of Common Olive leaf) /CF
5. Record the two Common Olive leaf surface area values in the appropriate table on the board.
6. Repeat steps 1-5 for the two leaves from the high sunlight habitat.
RESULTS & WRITE-UP
Each student will prepare an individual report using the class data. This report should follow the scientific
report format as presented by your instructor and described in the report guidelines and graphing data on
pages 114-115 of the lab manual.
114
SCIENTIFIC WRITTEN REPORT GUIDELINES
Each student must write his/her individual report, even though the experiments were conducted as a group.
Reports that do not appear to be original work will receive zero credit. The report should follow the style described
below.
Write the report in the past tense ALL the way through.
INTRODUCTION
In this section, briefly describe the background of your project. This should include:
 Background information that explains why you made your specific prediction.
 What was the idea or question that motivated your research?
 What was your working hypothesis? Remember that a hypothesis is a statement of prediction and is
often written as an “if...then” prediction statement.
MATERIALS & METHODS
In this section, succinctly describe what you did to set up and conduct your experiment.
Give basic instructions, a step-by-step guide which would allow someone to accurately repeat your experiment on
their own. DO NOT list the materials used; mention them when needed when describing the methods.
RESULTS
In this written section, you report the outcome of your research (the data) in a paragraph. This should be just the
facts of what you saw or measured including any trends or patterns that can be observed in your data. DO NOT
make conclusions or relate your data to your hypothesis here. In addition to the paragraph of results, you must also
add table(s) and graph(s) of your data. These can be referred to in the proper place in your results and/or your
discussion sections (e.g., Table 2, see Figure 1, etc.).
Each graph must contain a figure number and a printed or typed descriptive caption.
Each data table must contain a figure number and a printed or typed descriptive caption.
DISCUSSION
This is the section where you get to give your opinions of the experiment and analyze the data. You interpret the
results and provide an explanation for the outcome of the experiment. Does it support or not support your
hypothesis? Were there any observable flaws or errors in the experiment that you need to explain? What other
factors could have affected your results? How would you do the experiment differently, if you wanted to improve
upon or expand your investigation? Did your results lead to any new questions? Are there any experiments you
would like to do as a result of your findings? Discuss data to support your statements.
FORMATTING INSTRUCTIONS
o Margins: 1 inch on all sides

o Font type: Times New Roman
o Font size: 12 point
o Line spacing: 1.5
o Name and Lab section Number: upper left corner
o Title: Below the lab section number, centered, bold, and all caps
o Page numbers: on the bottom (right side or centered)
o Section headings: bold and all caps
o Total length: Maximum of TWO typed pages not counting tables and graphs
115
GRAPHING DATA
Graphing is a way of turning numbers (data) into pictures. This helps in recognizing
trends or patterns in the data. There are three basic types of graphic forms. Each is useful for
comparing different types of information.
PIE GRAPH
This type of graph is used when you want to compare parts to a

whole. It is often used with percent (comparing a part to the
total) as with money as in a budget (individual expenses to the
total budget).
BAR GRAPH:
This type of graph is used when you want to compare groups

or categories with each other. The vertical (y) axis is a linear
sequence starting at the bottom at zero. The categories to be
compared are lined up on the horizontal (x) axis.
3. LINE GRAPH:
This type of graph is used when the variables being studied

are linear (going from low to high, slow to fast, young to old,
etc.). This is often the most useful graph in science because
many of the variables studied are linear. In a line graph the
intersection of the axis represents the zero point (or the
smallest number). Both the y-axis and the x-axis move away
from this point as linear lines. The units should be equally Trend line
spaced as in a number line. Remember that the purpose of a
graph is to indicate trends: after the data points are plotted, a
trend line is drawn. When drawing this line you do not
connect the data points but rather draw a straight line that best
fits all the data.
116
SCIENTIFIC WRITTEN REPORT GRADING POLICY

 Scientific written lab reports are 20 points each.
 These reports are due the next lab session after each experiment was completed.
 Late lab reports are not accepted!
 The reports must be written in the past tense ALL the way through.
1. INTRODUCTION:
a. Background information ........................................................................................... 1 point
b. The question .......................................................................................................... 1.5 points
c. The hypothesis ...................................................................................................... 1.5 points
2. MATERIALS AND METHODS:

a. Paragraph of Methods with Materials incorporated into it ....................................... 5 points
3. RESULTS:
a. Paragraph of results .................................................................................................. 2 points
b. Graph(s) .............................................................................................................. 1.75 points
Graph(s) : each must contain a figure number and a printed or typed descriptive caption..0.25 point
c. Table(s) ................................................................................................................ 0.75 point

Data table(s): each must contain a figure number and a printed or typed descriptive caption. 0.25 point.
4. DISCUSSION:
a. State if the results refuted or supported the hypothesis ............................................. 1 point
b. State your opinion (were there any observable flaws or errors in the experiment
that you need to explain? What other factors could have affected your results?
How would you do the experiment differently, if you wanted to improve upon or
expand your investigation?) ...................................................................................... 1 point
c. Discussion of data to support the above (a & b) ............................................ …… 3 points
Formatting Instructions (1 point total)

o Margins: 1 inch on all sides………………………………………………………….0.1 point
o Font type: Times New Roman……………………………………………………….0.1 point
o Font size: 12 point…………………………………………………………………....0.1 point
o Line spacing: 1.5……………………………………………………………………..0.1 point
o Name and Lab section Number: upper left corner…………………………………..0.1 point
o Title: Below the lab section number, centered, bold, and all caps………………….0.1 point
o Page numbers: on the bottom (right side or centered)……………………………….0.1 point
o Section headings: bold and all caps…………………………………………………0.2 point
o Total length: Maximum of TWO typed pages not counting tables and graphs.…... 0.1 point
Half a point (0.5 point) will be deducted from your score for each of the following:
a. Report not written in past tense ALL the way through

b. Poor spelling
c. Poor grammar
117

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Topics in Biology

INTRODUCTION TO BIOLOGY 100 LABORATORY ............................................................................ 3

BIOLOGICAL CLASSIFICATION ........................................................................................................... 16

DIFFUSION AND OSMOSIS .................................................................................................................... 34

DNA: REPLICATION, TRANSCRIPTION, & TRANSLATION ............................................................ 42

MITOSIS AND MEIOSIS………………………………………………………………………………..49

MENDELIAN GENETICS ........................................................................................................................ 58

ORGAN SYSTEMS ................................................................................................................................... 68

PLANT ANATOMY .................................................................................................................................. 83

STARCH DIGESTION EXPERIMENT………………………………………………………………..103

OSMOSIS EXPERIMENT ....................................................................................................................... 106

GROWTH OF BACTERIA EXPERIMENT ............................................................................................ 108

ALLELOPATHY EXPERIMENT ........................................................................................................... 110

VEGETATION ANALYSIS EXPERIMENT .......................................................................................... 113

SCIENTIFIC WRITTEN REPORT GUIDELINES ................................................................................. 115

GRAPHING DATA .................................................................................................................................. 116

SCIENTIFIC WRITTEN REPORT GRADING POLICY ....................................................................... 117

INTRODUCTION TO BIOLOGY 100 LABORATORY

INTRODUCTION TO BIOLOGY LABORATORY TECHNIQUES

8. Before returning your microscope to your storage cabinet:

3. Open the iris diaphragm approximately half way.

4. Turn the light source to the lowest setting.

EXERCISE 1: Identify the following Microscope Parts (1 point)

EXAMINING SPECIMENS WITH THE MICROSCOPE

EXERCISE 2: Examination of Colored Threads (1 point)

1. Obtain a slide of colored thread.

3. Draw the thread at 40X, 100X and 400X total magnification.

1. Which color thread is on top? ____________ Middle? ____________ Bottom? ____________

OBJECTIVE TOTAL MAGNIFICATION

EXERCISE 3: Examination of the Letter “e” (1 point)

EXERCISE 4: Preparation and Examination of Wet Mounts (2 points)

5. SLIDE CLEANUP PROCEDURE

EXERCISE 5: Introduction to Other Laboratory Materials (1 point)

Metric Units of Measurement

Quantity Metric Unit Symbol

Some Divisions of Metric Units

Prefix Symbol Value Prefix Symbol Value

Volume of test tube = _______ ml

Length = ________ mm Diameter = _______ mm

Weight = ________ g Length= _______ mm

EXERCISE 6: Observations…questions…hypotheses… (4 points)

BOX OBSERVATION SHEET #1 (2 points) Name

Technique Observations Conclusion

What did you The actual What does this

Total of your conclusions: What do you think the object is?____________________

Questions (1 point each):

Turn in pages 3-15 for this lab.

Below is an example of a simple analytical key.

1. a. object is spherical ......................................................... 2

2. a. object is less than 4 cm in diameter ........................grape

3. a. object is orange in color .......................................orange

4. a. object is yellow in color ....................................... lemon

Any characteristic can be used.

The exercises in this lab are based on the traditional taxonomy.

EXERCISE 1: Make a Dichotomous Key (Part I) and

Questions (0.5 points each):

EXERCISE 2: Make a Dichotomous Key (Part II) (2 points)

EXERCISE 3: Make a Dichotomous Key (Part III) and

1. Organize the plastic bugs into ten groups or piles.

Questions (0.25 points each):

1. What were your observations (list 2 or more)?

2. What was your question (list 2 or more)?

3. What was your hypothesis?

4. Did you test your hypothesis? How?

1. Which color thread is on top? Middle? Bottom?

Length = __ mm Diameter = _ mm

Weight = __ g Length= _ mm

1. Which molecule diffused the fastest? ___ The slowest? _

1. Which solution(s) is hypotonic to blood cells? __ Hypertonic?

1. Which solution is hypertonic to the plant cells? Hypotonic?