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
MEMBRANE is from the region of higher
TRANSPORT concentration to a region of lower
concentration. The process continues
until the substance is equally distributed
throughout the medium.
Osmosis is a special form of
diffusion. It is the 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 1% potassium permanganate Distilled water

Iron stand with clamp 100mL beaker Paper towels
Cotton balls Thermometer Glass slides
Ruler Ice Cover slips
1M HCl Potassium permanganate crystals 0.9% NaCl solution
1M NH4OH 50mL beaker 5% NaCl solution
Petri dish with agar Iodine solution Blood
Cork borer 1% starch solution Microscope
1% Congo red Cellulose sac
1% methylene blue 0.2M-1M sucrose solution

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 NH 4OH (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 water and inform the teacher immediately.

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).

Figure 1. Agar plate with holes

2. Fill each hole with equal amounts of one of the following solutions: 1% Congo
Red, 1% Methylene Blue, 1% Potassium Permanganate. Do not let the
solutions overflow.

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 temperatures.

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

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 50mL 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 semipermeable

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

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

2. Fill the sacs with 10mL 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 250mL 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 minutes.

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,
distilled water, 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