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Polytechnic University of the Philippines

College of Social Sciences and Development


Department of Psychology

BIOLOGICAL MEMBRANES
DIALYSIS, OSMOSIS AND DIFFUSION

Abstract
This laboratory experiment is meant to be done for the students to be able to
understand and find the similarities and differences of Dialysis, Osmosis and Diffusion in
Biological Membranes and how they relate and connect to each other. This experiment
will show the relationship of temperature and concentration in molecular level and
through biological membranes. The main objective of performing the experiment is to
show the effects of concentration and temperature on diffusion and osmosis. This
experiment will help us know the importance of diffusion and osmosis to cells
particularly to animal and plant cells.
KEYWORDS: 1) Dialysis, 2) Osmosis, 3) Diffusion, 4) Biological membranes, 5)
Temperature and concentration, 6)Molecular level, 7) Animal and plant cells

Introduction
All cells are all like living organisms inside our bodies. They are the one that is
responsible to the things happening inside and outside of the body. Just like us, cells
have a protection. All cells are enclosed with a cell membrane. Each cell membrane in
each cells have a function. A cell membrane is like a barrier that allows the passage of a
liquid or substance to enter or leave a cell.
Diffusion, osmosis and dialysis, as we all know are widely known concepts in
both Biology and Chemistry. These concepts are widely represented in many books in
many ways to be able to help students analyze and understand how it works, especially
at molecular levels. In order to explore and familiarize these concepts, we performed an
experiment to end up with conclusions relevant to the main objective of the experiment
which is to show effects of concentration and temperature on diffusion and osmosis
There are processes in which how the cell membrane allows the passage of a
substance in the cell the diffusion, osmosis and dialysis. Osmosis is the process that
causes a liquid (especially water) to pass through the wall of a living cell (MerriamWebster, 2016). Diffusion refers to the process by which molecules intermingle as a
result of their kinetic energy of random motion (BBC, 2014). The researchers learn how
cell membrane functions in different ways. The researcher conducted an experiment
which allows to know how diffusion, osmosis and dialysis works within a cell. This
experiment also showed types of passive transport namely, diffusion and osmosis. The
experiment will show the movements of molecules. It also shows us how the hypertonic,

isotonic and hypotonic solutions reacted when isotonic saline solution, hypotonic salt
solution and hypertonic salt solution reacted to red blood cells.

Methodology
1. Diffusion
1.1.

Diffusion of solutes in water

A dot of
KMnO4

Place in each
test tubes

Distilled
Water

1.2.

Tap Water

Hot
Distilled
Water

Cold
Distilled
Water

Test tube 3

Test tube 4

Diffusion in a colloid

Melt gelatine

Test tube 1

Test tube 2

Allow to
cool

Add ~0.5ml of KMnO4,


Methylene Blue and Congo Red

Measure the migration of the dye


starting at the top-level of the
gelatine for 120mins.

Repeat
procedure for
set 2

2. Osmosis
2.1.

Osmosis (Longganisa Casting Model)

Make a solution of
~10ml sugar and
water

Repeat
procedure for
set 2

2.2.

Place on a
test tube

Obtain the weight of the


test tube at 10-mins
interval for 60-mins.

Cover the mouth of the


test tube with 1-ply of
Langgonisa casing and
secure it with rubber
band.

Weigh

Suspend the test tube


upside down in a 250ml beaker containing
water. Use iron clamp to
hold the test tube.

Osmosis in RBC

Drop blood in
each slides

Slide 1

Apply a drop of
Hypotonic Solution
(0.30 M NaCl)

Slide 2

Apply a drop of
Hypertonic Solution
(3.0M NaCl)

Slide 3

Apply a drop of
Isotonic-Normal
Solution (0.85M NaCl)

Observe under
microscope

3. Dialysis
Fill test tube with 3ml
Methylene Blue and 5ml
Saturated CaCl2 Solution

Cover the mouth of the


test tube with 1-ply of
Langgonisa casing and
secure it with rubber
band.

Tabulate the
results

Suspend the test tube


upside down in a 250ml beaker containing
water. Use iron clamp to
hold the test tube.

Test the presence of


Methylene Blue,
Chloride and and
Calcium in the beaker
after every minute in
30-mins.

The experiment was divided into 3 parts. First diffusion and is divided into 2 subparts; diffusion of solutes in water and diffusion in a colloid. The second one, osmosis
and was also divided into 2 sub-parts; osmosis (longganisa casing model) and osmosis
in RBC. The last part is, dialysis.
The first process is, Diffusion. A series of test tubes containing distilled water, tap
water, hot distilled water (70 OC or higher) and cold distilled water on ice were
separately prepared. The test tubes were labelled accordingly. After labelling, a dot
amount of KMnO4 crystals were placed to each test tubes and took note of the time at
which the KMnO4 crystals were completely dissolved in the water.
The second process is diffusion in colloid. In the experiment of diffusion of
colloid, the gelatin was melted according to the manufacturers procedure. Next,
prepare 8-10 test tubes and pour the gelatin mixture to each test tube and allow it to
solidify. Two sets of gelatin were made. After the gelatin was completely hardened,
overlay 0.05 ml of the potassium permanganate, Congo red and Methylene blue in each

test tube. Finally, measure the migration of the dye starting at the top level of the gelatin
using the ruler and record the measurements for 60 minutes at 10 minutes intervals or
until the dye reaches the bottom.
For the first process of osmosis, which is the longganisa casing model, the first
thing to do is to place 10 mL of saturated sugar solution in a test tube. Next, cover the
mouth of the test tube with 1-ply of longganisa skin and secure it with a rubber band.
After securing the cover of the test tube, use a triple beam balance to acquire the
weight of the covered test tube. Next, suspend the test tube upside down in a 250 mL
beaker containing distilled water and used and iron clamp to fasten the test tube. Lastly,
obtain the weight of the test tube at 10-minute intervals for 60 minutes and tabulate the
data and observations.
The second process in osmosis is observed by the presence of RBC. The first
step is to prepare 3 clean glass slides and cover slips with a drop of blood on each
glass slides. You can use sterile lancets to prick a donors finger to be able to get
samples of RBC. After that, apply drop of Isotonic- Normal Saline Solution (0.85 M
NaCl) in one glass slide, a drop of hypertonic salt solution (3.00 M NaCl) in another
glass slide and hypotonic salt solution (0.30 M NaCl) to the remaining glass slide. To be
able to end up with a conclusion, observe the appearance and sizes of the RBCs under
the microscope.
The last part of the experiment is, dialysis. The first step in this experiment is to
fill a test tube with 3 mL Methylene blue solution and 5 mL saturated CaCl2 solution.
Cover the test tube with 1-ply of longganisa casing and secure it with a rubber band.

After that, suspend the test tube upside down in a 250 mL beaker containing distilled
water and use an iron clamp to fasten it. The main objective of this experiment is to test
the presence of Methylene blue, chloride and calcium. We can test this by taking a
sample of beaker-water for 30 minutes in 2-minutes interval. Lastly, tabulate the results
and observations.

Results
1. Diffusion
1.1.
Diffusion of Solutes in Water
TABLE 1. Time consumed to dissolve Potassium Permanganate
Time where KMnO4

Distilled Water
Tap Water
Hot Distilled Water
Cold Distilled Water

completely dissolved in

Ranking

water
1 hour and 10 minutes
1 hour and 16 minutes
40 minutes
50 minutes

Third
Fourth
First
Second

In the table shown in the results, Table 1, of how much time the four waters
consumed to dissolve the KMnO4. The hot distilled water dissolved the KMnO 4 for 40
minutes which is the fastest between the four. While, tap water has the longest time to
dissolve the substance in 1 hour and 16 minutes.
1.2.

Diffusion in a Colloid
TABLE 2. SET A: Migration of the three substance in a gelatin

Chemical

10min

10min

10min

10min

10min

10min

10min

10min

10min

10min

s
KMnO4
Congo

s
0.5cm
0.5cm

s
0.5cm
0.5cm

s
0.6cm
0.8cm

s
0.6cm
0.9cm

s
NC
1.0cm

s
0.6cm
NC

s
NC
NC

s
NC
1.3cm

s
0.6cm
NC

s
NC
1.4cm

Red
Methylen

0.5cm

0.5cm

NC

NC

NC

0.6cm

NC

NC

0.7cm

NC

e Blue

TABLE 3. SET B: Migration of the three substance in a gelatin


Chemical

10min

10min

10min

10min

10min

10min

10min

10min

10min

10min

s
KMnO4
Congo

s
0.5cm
0.5cm

s
NC
NC

s
0.6cm
0.8cm

s
0.6cm
0.9cm

s
NC
1.0cm

s
0.6cm
NC

s
NC
NC

s
NC
1.3cm

s
0.6cm
NC

s
NC
1.4cm

Red
Methylen

0.5cm

NC

NC

NC

NC

0.6cm

NC

NC

0.7cm

NC

e Blue

In the two tabulated results for Set A and B (Table 2 and 3), all three chemical
migrate in the colloid. Congo red migrated the largest amount of 1.4cm for both Set A
and B. While, KMnO4 shows the least migration in all three chemicals with only 0.6cm in
both Set A and B.
2. Osmosis
2.1.
Osmosis (Longganisa Casing Model)
TABLE 4. Decrease of weight of the Langgonisa Casing Model

Starting Weight
10 mins
20 mins
30 mins
40 mins
50 mins
60 mins

Set A
41.2g
41.9g
41.5g
39.1g
38.8g
36.4g
35.9g

Set B
30.0g
30.4g
30.1g
29.9g
29.3g
28.9g
28.2g

For 60 minutes, the researchers obtain the weight of the set-up for each 10
minutes. The tabulated data, Table 4, shows that the weight of the set-up decreases in

every 10 minutes interval. This shows that in every 10 minutes, the sugar solution inside
the test tube decreases.
2.2.

Osmosis in RBC

FIGURE 1. FIGURE 2.
FIGURE 3.
RBC with
RBC with
RBC with
The Hypertonic salt solution turned the RBC to steadily increase its pressure and
Hypertonic SaltHypotonic Salt
Normal-Saline
Solution size that
Solution
Solution
made it more visible than the
RBC with Hypertonic salt solution. However, the
FIGURE 1.
RBC with RBC with a drop of Normal-Saline Solution retains its normal size.
Hypertonic Salt
Solution
3. Dialysis
TABLE 5. Changes in the water every minute
Time

2 mins

Oxalic Acid
No reaction

Silver Nitrate
There is a change

Methylene Blue
The water in the

in color in the first

beaker is still clear.

try. The water in the


test tube became

4 mins
6 mins

slightly blurry wihite.


The color of blurry

white still continues


There are tiny
to darken.
precipitates are can
be seen in the
8 mins

mixture and the


blurry white color
became more

10 mins

12 mins
Little formation of
moist in the test
14 mins

tube wall and a mild


change in

obvious.
As the time goes

A touch of color bue

on, the blurry white

isa can be seen on

color became

the water in the

darker and the

beaker but in a

precipitate became

slightly manner.
As the time goes

more obvious.

on, the color of the


Methelyne Blue
became more
obvious in the water

temperature
The moist formed

in the beaker. It

16 mins

more in the wall of

grows darker and


darker.

18 mins

the test tube


The moist become
more obvious
compared to the

earlier minutes and


the temperature
continue to grow
colder
The temperature
started to decrease
and become colder
20 mins
and the moist is still
growing more in the
22 mins
24 mins
26 mins

test tube glass


As the time goes
on, the temperature
in the test tube
decreases that it

28 mins

grows colder and


the moist is become
more noticeable
The precipitate
The moist in the test
formed became
tube became more
more obvious that it

The color blue in

floats in the mixture.

the water became

The blurry white

darker than the

color is at its higher

earlier minutes.

noticeable that it
30 mins

almost made the


glass blurry. The
temperature
compare to the
remains cold.
earlier colors.

The tabulated results in Table 5 shows the time where each mixtures made their
reaction. This clearly displays that Silver Nitrate pass through the langgonisa casing first
in 2 minutes. While, Oxalic Acid is the last mixture that shows reaction that in 14
minutes.
Discussion
1. Diffusion
1.1.
Diffusion of Solutes in Water
The experiment anticipates to know of the four temperatures or kinds of water
can dissolve KMnO4 faster.
The tap water, distilled water, hot distilled water and cold distilled water act as a
solvent in which they have to dissolve Potassium Manganate (KMnO 4). In the set-up,
there are 4 test tubes that are placed in the test tube rack. Each of the test tubes
contains the same amount of the four waters. The researchers drop a small amount of
KMnO4 in each test tube.
In the table shown in the results, Table 1, of how much time the four waters
consumed to dissolve the KMnO4. The hot distilled water dissolved the KMnO 4 for 40
minutes which is the fastest between the four. While, tap water has the longest time to
dissolve the substance in 1 hour and 16 minutes.
1.2.

Diffusion in a Colloid

The experiment is expecting to learn which chemical will make a larger migration
in a colloid by using KMnO4, Methylene blue and Congo Red.
Colloids are substance that consists of particles dispersed throughout another
substance which are too small (Merriam-Webster, 2016). In this experiment, the
researchers used a gelatin to form a colloid. The researchers drop an approximately
0.5ml of each chemical in each of the 3 test tubes.
In the two tabulated results for Set A and B (Table 2 and 3), all three chemical
migrate in the colloid. Congo red migrated the largest amount of 1.4cm for both Set A
and B. While, KMnO4 shows the least migration in all three chemicals with only 0.6cm in
both Set A and B.
2. Osmosis
2.1.
Osmosis (Longganisa Casing Model)
This experiment goals to know the weight of the set-up of test tube with sugar
solution and a longganisa casing as its cover for every 10 minutes in 60 minutes.
The role of the langgonisa casing is to act as a semi-permeable membrane that
limits the passage of water in each container test tube and beaker. There are two
pressures that are can be found in this set-up, the one that is coming from the test tube
and the one that is coming from the water in the beaker.
For 60 minutes, the researchers obtain the weight of the set-up for each 10
minutes. The tabulated data, Table 4, shows that the weight of the set-up decreases in
every 10 minutes interval. This shows that in every 10 minutes, the sugar solution inside
the test tube decreases.

2.2.

Osmosis in RBC

In this experiment, the researchers are assigned to define which is the biggest
Red blood cell may form when being dropped by Hypotonic Salt Solution, Hypertonic
Salt Solution and Normal-Saline Salt Solution.
The Hypertonic Salt Solution concentration of water is higher within the cell.
There is a net movement of water from inside to outside of the cell. Water leaves the
cell by osmosis which causes the cell to shrink as its internal pressure decreases.
Hypotonic Salt Solution contains higher concentration of water and lower concentration
of solutes. Since, the concentration of water is higher outside the cell. There is a net
movement of water from outside into the cell. Cell gains water, swells and the internal
pressure increases. Eventually burst (Haemolysis). While, Isotonic solution is a solution
in which the concentration of solutes are equal, so: first, water diffuses in and out of the
cell at equal rates; second, here is no net movement of water across the plasma
membrane; third, the cells retain their normal shape.
The Hypertonic salt solution turned the RBC to steadily increase its pressure and
size that made it more visible than the RBC with Hypertonic salt solution. However, the
RBC with a drop of Normal-Saline Solution retains its normal size.
3. Dialysis
The experiment expects to know which of the three chemical (Oxalic Acid, Silver
Nitrate and mixture of Methylene Blue and Saturated CaCl 2) will pass through the
langgonisa casing first.

Langgonisa casing acts as a semi-permeable membrane a barrier that allows


some molecules to move through it, where in others cannot. In the set-up that is used
where the test tube is being suspended in the water inside the beaker while it is
positioned upside-down, there are two pressures acting the pressure inside the test
tube and the one in the beaker, which controls the exchange of mixture inside and out of
the test tube. The mixture of Methylene blue and Saturated CaCl 2 is passing through the
langgonisa casing which affects the water in the beaker that it turns into blue.
While doing the experiment, the researchers made 2 sets of testubes, one test
tube is for the Oxalic Acid and the other one is for the Silver Nitrate. In every 2 minutes
in 30 minutes, the researchers gathered small amounts of water in the beaker and pour
it into each of the test tubes. The researchers continued this process for 30 minutes.
The tabulated results in Table 5 shows the time where each mixtures made their
reaction. This clearly displays that Silver Nitrate pass through the langgonisa casing first
in 2 minutes. While, Oxalic Acid is the last mixture that shows reaction that in 14
minutes.

Reference
BBC. (2014). Diffusion. BBC-GCSE Bitesize: Diffusion. BBC Teachers, 2014.
http://www.bbc.co.uk/schools/gcsebitesize/science/add_aqa_pre_2011/cells/cells
3.shtml
Beltran, Agapito J., et al. (2005). Biology 22: General Zoology Laboratory Manual (Third
Edition).
Blamire, John. (1999). Osmosis: Brother Gregory Investigates.
http://www.brooklyn.curry.edu/bc/ahp/ChemInvest/CI.Q2.html.
Merriam-Webster Dictionary. (2016). Colloid.
Merriam-Webster Dictionary. (2016). Hypertonic.
Merriam-Webster Dictionary. (2016). Hypotonic.
Merriam-Webster Dictionary. (2016). Isotonic.
Merriam-Webster Dictionary. (2016). Osmosis.
Verma, Rituraj. (2016). What is the difference between diffusion and osmosis?.
https://www.quora.com/What-is-the-difference-between-diffusion-and-osmosis