Polytechnic University of the Philippines

College of Social Sciences and Development

Department of Psychology

BIOLOGICAL MEMBRANES
(DIALYSIS, OSMOSIS AND DIFFUSION)

Submitted by:
SAN MIGUEL, Djanelle Mei M.
DELOS REYES, Erica Gabrielle
MORAL, Chatlette Karl
COLAMBO, Mark John Peter
GO, Thea
CABIGAO, Jereme Anne

Submitted to:
Prof. Cris Argamino

BS PSYCHOLOGY 3-4

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 knowing how diffusion, osmosis and dialysis work 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

Place on a
test tube

Repeat
procedure
for set 2

Obtain the weight of the

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

2.2.

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
Slide 1

Drop blood
in each
slides

Slide 2

Slide 3

Apply a drop of
Hypotonic Solution
(0.30 M NaCl)
Apply a drop of
Hypertonic Solution
(3.0M NaCl)
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 two
sub-parts; diffusion of solutes in water and diffusion in a colloid. The second one,
osmosis and was also divided into two sub-parts; osmosis (longganisa casing model)
and osmosis in red blood cell (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 C or higher) and cold distilled water on ice were
separately prepared. The test tubes were labelled accordingly. After labelling, a dot
amount of potassium permanganate (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 use 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 three 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 two 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
completely dissolved in

Ranking

water
Distilled Water

1 hour and 10 minutes

Third

Tap Water

1 hour and 16 minutes

Fourth

Hot Distilled Water

40 minutes

First

Cold Distilled Water

50 minutes

Second

Table 1 shows how much time the four waters dissolved KMnO4. The hot distilled
water dissolved KMnO4 in about 40 minutes which is the fastest among 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

KMnO4

0.5cm

0.5cm

0.6cm

0.6cm

NC

0.6cm

NC

NC

0.6cm

NC

Congo

0.5cm

0.5cm

0.8cm

0.9cm

1.0cm

NC

NC

1.3cm

NC

1.4cm

0.5cm

0.5cm

NC

NC

NC

0.6cm

NC

NC

0.7cm

NC

Red
Methylen
e Blue
*Note: NC means no change.
TABLE 3. SET B: Migration of the three substance in a gelatin
Chemical

10min

10min

10min

10min

10min

10min

10min

10min

10min

10min

KMnO4

0.5cm

NC

0.6cm

0.6cm

NC

0.6cm

NC

NC

0.6cm

NC

Congo

0.5cm

NC

0.8cm

0.9cm

1.0cm

NC

NC

1.3cm

NC

1.4cm

0.5cm

NC

NC

NC

NC

0.6cm

NC

NC

0.7cm

NC

Red
Methylen
e Blue
*Note: NC means no change.
In the two tabulated results for Set A and B (Table 2 and 3), all three chemical
migrated in the colloid. Congo red migrated the largest amount of 1.4 cm for both Set A
and B, while, KMnO4 shows the least migration in all three chemicals with only 0.6 cm in
both Set A and B.
2. Osmosis
2.1.

Osmosis (Longganisa Casing Model)

TABLE 4. Decrease of weight of the Langgonisa Casing Model

Set A

Set B

Starting Weight

41.2 g

30.0 g

10 mins

41.9 g

30.4 g

20 minutes

41.5 g

30.1 g

30 minutes

39.1 g

29.9 g

40 minutes

38.8 g

29.3 g

50 minutes

36.4 g

28.9 g

60 minutes

35.9 g

28.2 g

For about 60 minutes, the researchers obtained the weight of each set- ups
every 10 minutes. The tabulated data, Table 4, shows that the weight of the set-up
decreases in every 10 minute intervals. This shows that in every 10 minutes, the sugar
solution inside the test tube decreases.
2.2.

Osmosis in RBC

FIGURE 1
RBC with
Hypertonic Salt
Solution

FIGURE 2.
RBC with
Hypotonic Salt
Solution

FIGURE 3.
RBC with
Normal-Saline
Solution

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
TABLE 5. Changes in the water every minute
Time

Oxalic Acid

Silver Nitrate

Methylene Blue

Theres a change in
color in the first try.
Two minutes

The water in the test

tube became
slightly blurry white.

Four minutes

The color of blurry

white still continues
The water in the

Six minutes
to darken.

beaker is still clear.

No reaction
There are tiny
precipitates that can
be seen in the
Eight minutes

mixture and the

blurry white color
became more
obvious.

10 minutes

As the time goes

A touch of color bue

on, the blurry white

can be seen on the

color became

water in the beaker

darker and the

but in a slightly

precipitate became

manner.

more obvious.

12 minutes
Little formation of
moist in the test
14 minutes

tube wall and a mild

change in
temperature
As the time goes
The moist formed
on, the color of the

16 minutes

more in the wall of

Methelyne Blue
the test tube
became more
The moist become
obvious in the water
more obvious
in the beaker. It
compared to the
grows darker and

18 minutes

earlier minutes and

darker.
the temperature
continue to grow
colder
The temperature

20 minutes

started to decrease
and become colder

and the moist is still

growing more in the
test tube glass
22 minutes

As the time goes

24 minutes

on, the temperature

26 minutes

in the test tube

decreases that it
grows colder and

28 minutes
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 the

floats in the mixture.

water became

The blurry white

darker than the

color is at its higher

earlier minutes.

noticeable that it
30 minutes

almost made the

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

The tabulated results in Table 5 show the time where each mixture made their
reaction. This clearly displays that Silver Nitrate pass through the longganisa casing first
in two minutes, while Oxalic Acid is the last mixture that shows reaction within 14
minutes.

Discussion
1. Diffusion
1.1.

Diffusion of Solutes in Water

The experiment anticipates knowing 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 four test tubes that are placed in the test tube rack. Each test tube contains
the same amount of four water solutions. The researchers drop a small amount of
KMnO4 in each test tube.
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
migrates in the colloid. Congo red migrates 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 aims to know the weight of the test tube set- ups with sugar
solution and a longganisa casing as it covers for every 10 minutes in 60 minutes.
The role of the longganisa 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 can be found in this set-up, one that is coming from the test tube and the
other that is coming from the water in the beaker.
2.2.

Osmosis in RBC

In this experiment, the researchers are assigned to define which red blood cell
can be formed the largest 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 concentrations 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
longganisa casing first.
Longganisa 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 longganisa casing which affects the water in the beaker that it turns into blue.
While doing the experiment, the researchers made two sets of test tubes, one
test tube is for the Oxalic Acid and the other one is for the Silver Nitrate. In every two
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.

Conclusion
To sum up, temperature, concentration gradient and molecular weights affect the
diffusion which is the movement of molecules from a region of higher concentration to
lower concentration. These transport processes got through the process of diffusion
across membranes which are semi-permeable allowing certain molecules to pass
through, regulating the entry and exit of molecules inside and outside the cell.
Temperature and concentration gradient varies directly to the rate of diffusion. As the
concentration gradient and temperature increases the rate diffusion will also
increase. On the other hand the molecular weight is inversely proportional to the rate of
diffusion.

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