The maintenance of a constant internal

environment in a cell or organism, also known as

homeostasis, necessitates a constant entrance of
materials into the cell and the passage of other
substances out of it. Materials are selectively
exchanged with the environment through the plasma
membrane.

3
The movement of substances through the plasma
membrane involves several processes. These include
passive transport or diffusion (simple and facilitated),
active transport and bulk transport (phagocytosis,
pinocytosis). Diffusion is the spontaneous movement of
molecules of different substances. It results from the
kinetic energy of the molecules. The movement of
molecules is from the region of higher concentration to
a region of lower concentration. The process continues
until the substance is equally distributed throughout the
MEMBRANE medium.
Osmosis is a special form of diffusion. It is the
TRANSPORT movement of water from a dilute solution (hypoosmotic)
to a more concentrated (hyperosmotic) one through a
semi permeable or selectively permeable membrane.
This type of membrane allows only certain molecules to
pass through. The process of diffusion of crystalloid
solute through such membrane on the other hand, is
known as dialysis.
This activity will familiarize the students with the
transport processes occurring within the cell.

Objectives At the end of this activity, the student

should be able to:

1. describe the processes involved in diffusion and

osmosis
2. relate molecular weight, temperature and molecular
size with the rate of diffusion
3. describe the role of the plasma membrane in the
transport process

Materials

Glass tube, iron stand with clamp, cotton balls, ruler,

1M HCl, 1M NH4OH
Petri dish containing agar, cork borer, 1% congo red,
1% methylene blue, 1% potassium permanganate,
ruler, 100 ml beaker, thermometer, ice, potassium,
permanganate crystals, 50 ml beaker, iodine solution,
1% starch solution, cellulose sac, 0.2 M-1M sucrose solution, distilled water, paper
towels, glass slides, cover slips, 0.9% NaCl, 5% NaCl solution, blood, microscope

Procedure

A. DIFFUSION

Diffusion and Molecular Weight

In this activity, the students will investigate the relationship of molecular weight and
diffusion rate.

Part 1
1. Obtain a clean glass tube. Clamp the tube horizontally to an iron stand.

2. Place a small piece of cotton ball in each end of the tube. The cotton must fill the
ends of the tube completely. Do not stuff the tube too full.

3. With a partner, simultaneously place 10 drops of 1M HCl (MW Cl=35) on the cotton
on one end of the tube and 10 drops of NH4OH (MW NH4+= 18) on the cotton at the
other end of the tube. Allow the cotton to be saturated. CAUTION : Do not spill
the chemicals, they may cause severe burns and may damage clothing. If
spilled on oneself or another person, flood the affected area with and inform
the teacher.

4. Observe the tube for a white ring that forms on the inside of the tube.

5. With a ruler, measure the distance from each end of the tube to the white ring.
Record your data in centimeter.

6. Rinse the tube with water and dispose the cotton in the designated container.

Part 2
1. Obtain a petri dish containing agar. Using a cork borer, carefully punch three holes
in the agar (Figure 1).

Fig.1 Agar Plate with holes

2. Fill each hole with equal amounts of one of the following solutions. Do not let the
solutions overflow.
1% congo red, 1% methylene blue, 1% potassium permanganate.

Note : The molecular weight of Congo red is 697, methylene blue is 319 and potassium
permanganate is 158.

3. After 30 minutes, examine the petri dish. Measure the diameters of the colored
rings around the holes in millimeters. Record your data on the Activity Sheet

Diffusion and Temperature

Temperature affects diffusion. In this section, a comparison will be made on the

rate of diffusion of potassium permanganate between two beakers having different
temperature.

1. Fill a beaker with half full of tap water. Place the thermometer into the beaker and
heat the water until the temperature is 50oC.

2. Fill another beaker about half full of ice water. Record the temperature.

3. Place both beakers in an area where they will not be disturbed. Slowly add same
amount (one crystal) of potassium permanganate in each beaker.

4. Over the next 30 minutes, observe the changes in the distribution of potassium
permanganate in each beaker.

5. Record the observations.

Diffusion and Molecular Size

In this part of the activity, the selectivity of a membrane will be demonstrated

based on the size of the diffusing molecule. A cellulose membrane will be used to
represent the cell membrane. The surface of the membrane contains tiny pores that
will allow certain molecules to pass through it but not the others. The experiment uses
starch and iodine solutions. The reaction between the two substances is indicated by
a blue-black color.

1. Fill one third of a 50 ml beaker with water. Then add 4 drops of IKI solution. Set
aside.

2. Obtain cellulose sac. Fill the sac with 1% starch solution. Securely tie the end of
the sac with a string.
3. Rinse the outside part of the sac under the tap. Make sure there are no
traces of starch outside the cellulose sac.

4. Place the sac inside the beaker prepared in #1.

5. Let stand for 20 minutes. Then check for any changes in color.

6. Record your observations.

B. OSMOSIS

Measuring the rate of osmosis

The rate of osmosis will be investigated using an artificial semi-permeable

membrane, the cellulose sac. The cellulose membrane is also known as dialysis
tubing.

1. Obtain 5 pcs. of 20- cm long cellulose sacs.

2. Fill the sacs with 10 ml of the designated solution:

Sac 1 – distilled water
Sac 2 – 0.4 M sucrose
Sac 3 – 0.8 M sucrose
Sac 4 – 1.0 M sucrose
Sac 5 - distilled water
3. Remove the bubbles inside the sac so it appears limp. Securely tie off the top end
of each sac with a string.

4. Wipe off excess water or solution from the sacs and weigh each sac using an
analytical balance. DO IT QUICKLY. Record the initial weight of each bag.

5. Fill 4 - 250 ml beakers with 2/3 full of distilled water. Fill one beaker with 2/3 full of
1M sucrose solution.

6. Immerse sacs 1-4 in separate beaker of distilled water, and sac 5 in a beaker of 1M
sucrose.

7. Let stand for 30 mins.

8. After 30 minutes, remove the sac from each beaker; wipe off excess water and
weigh. Record the weight as final weight.

9. Compute the percent change in weight using the formula below:

change in weight = final weight – initial weight X 100%
initial weight

10. Graph the change in weight against the concentration of the sucrose.
Osmosis and Red Blood Cells
This part of the activity will allow the students to observe osmosis in Red Blood
Cells. One student per group will be asked to give blood for observation.

1. Clean and dry three slides and three coverslips. Label the slides 0.9% NaCl, DW
and 5% NaCl.
2. Wash hands and prepare the sterile lancet or the pricking device
3. Choose either the middle finger or the ring finger of the non-dominant hand and
gently massage the finger from the base to the tip several times.
4. Wipe fingertip with alcohol swab. Dry thoroughly before pricking.
5. Position lancet opening against the fingertip and press the release lever until it
clicks.
6. Gently squeeze the finger from the base to obtain large drop of blood. Apply the
drop onto the labeled slides.
7. Put a drop of the respective solution on each slide. Mix the blood and the solution
with a toothpick, and place a coverslip. Examine the size and shape of the cells
under high power objective. DO NOT ALLOW THE SOLUTIONS TO DRY UP. This
will change their concentrations.
8. Compare the appearance of the cells on slides A, B and C.

References

Bailey, P.C., Hollman, D.C, Quarles, T. S. and Waits, E. D. 1970. Laboratory Guide
for An Introduction to Modern Biology. International Textbook Co. 111-118.

Feldman, S. 1965. Experiments in Biological Design. Holt, Rinehart and Winston, Inc.
USA 95-96.

http://web.ukonline.co.uk/webwise/spinners/life/osmosis.htm.

http://ekcsk12.org/science/lelab/membraneslab 1.htm1

http://www.accessexcellence.org/atg/data/released/0081-JeffLukens/index.htm1

http://biology.arizona.edu/sciconn/lessons/mccandless/
reading.htm1
 Activity 3
Cell Transport
Name __________________________________ Date Performed______________________
Course/Section___________________________ Date Submitted ______________________
Instructor ________________________________

Diffusion and Molecular Weight

Part 1
Distance covered by :

HCl : _________ cm NH4OH : __________cm

1. What compounds are formed upon the reaction of HCl and NH4OH?

2. How does molecular weight affect the rate of diffusion?

Part 2
Diameters of the rings:

Congo red _____ mm Methylene blue ____ mm

Potassium permanganate _____ mm

1. Which solution has diffused the farthest? _____________________________

2. How does molecular weight affect the rate of diffusion?

__________________________________________________________________
__________________________________________________________________
_________________________________________________________________

Diffusion and Temperature

1. Why is potassium permanganate used in this experiment?

________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
2. Describe the distribution of potassium permanganate in each beaker after 30
minutes.
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________

3. What is the effect of temperature on the rate of diffusion? Explain.

__________________________________________________________________
__________________________________________________________________
__________________________________________________________________
__________________________________________________________________
__________________________________________________________________

Diffusion and Molecular Size

1. Where did you observe a color change, in the beaker or cellulose sac? How do
you explain the color change?
__________________________________________________________________
__________________________________________________________________
__________________________________________________________________
__________________________________________________________________
__________________________________________________________________

2. Based on the results of your experiment, which substance has smaller molecules,
starch or iodine? Explain your answer.

________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
Measuring the rate of osmosis

Initial Weight of Final Weight of % change in

Set-Up cellulose sac cellulose sac Weight

Sac : distilled water

Beaker : distilled water
Sac : 0.4 M sucrose
Beaker : distilled water
Sac : 0.8M sucrose
Beaker : distilled water
Sac : 1.0 M sucrose
Beaker : distilled water
Sac : distilled water
Beaker : 1.0M sucrose

Graph

0.0

1. What is the relationship of the rate of osmosis with the concentration of sucrose on
one side of the membrane (concentration gradient of sucrose)?
__________________________________________________________________
__________________________________________________________________
__________________________________________________________________

2. Explain the results in sac #5.

__________________________________________________________________
__________________________________________________________________
__________________________________________________________________
__________________________________________________________________
3. In what way does the cellulose sac/dialysis sac similar to the cell membrane?

________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________

Osmosis and the Red Blood Cell

1. Describe the shape and size of red blood cell seen on the three slides. Draw your
observations.

Slide A: _____________________________________________________________

Slide B: _____________________________________________________________

Slide C: _____________________________________________________________

2. Based on the results of your experiment,

2.1. Which solution is hyperosmotic relative to the red blood cell? Explain.
__________________________________________________________________
__________________________________________________________________
__________________________________________________________________

2.2. Which solution is hypoosmotic relative to the red blood cell? Explain.
__________________________________________________________________
__________________________________________________________________
__________________________________________________________________

2.3. Which solution is isosmotic relative to the red blood cell? Explain.
__________________________________________________________________
__________________________________________________________________
__________________________________________________________________