Experiment 6

EXPERIMENT 6
Membrane Structure and Function

(Tonicity and Diffusion)
INTRODUCTION
In order for cells to interact with their environment, chemicals, including water, must be able to
move across the cell membrane and across the cell. Movement within the cell occurs by a
process known as diffusion. Molecules move across the cell membrane by a related process
known as osmosis. Diffusion is the movement of molecules from a region of higher
concentration to a region of lower concentration. This happens because of random molecular
motion. Molecules move around randomly until there is an even mixture throughout the
container in which they are enclosed. The overall effect is that molecules move "down" a
concentration gradient from a region of high concentration to a region of low concentration.
Osmosis is the movement of water down a concentration gradient and at the same time across a
membrane. Cell membranes do not allow all molecules to cross them. They are said to be
"selectively" or "differentially" permeable. Only certain molecules can cross the membrane into
or out of a cell. For example, water can cross the membrane while sodium and chlorine ions
(dissolved salt) cannot. If there is a concentration gradient across the membrane (if there is more
salt and less water on one side than on the other), water will move across the membrane down
the concentration gradient while the salt cannot. If there is more salt and less water inside a cell
than outside, water will flow into the cell from the surrounding environment. This process is
called osmosis.
When the environment outside a cell has a lower concentration of dissolved molecules than
inside the cell, the solution is said to be hypotonic, and water will move from the solution into
the cell. If the surrounding solution has a higher concentration of dissolved molecules than the
cell, the solution is hypertonic. In that case, water will move from the cell out into the
surrounding solution. An isotonic solution is one in which the concentration of dissolved
molecules is the same inside and outside the cell, and there is no net movement of water across
the membrane. When cells are placed in a hypertonic solution, water flows out of them and they
shrink or shrivel up. When cells are placed in a hypotonic solution, water flows into them. If the
cell does not have a cell wall or some other means of protecting the membrane, it will burst in a
hypotonic solution.
MATERIALS
Potato strips Test tubes 10% NaCl
Distilled water Agar Petri Dish Timer
Potassium Permanganate (KMnO4) Ruler 0.9% NaCl
20 % NaCl Red Blood Cells Onions
Slides and cover slips Microscope
METHOD
A. Tonicity in Potato strips:
1. Cut two strips of potato, each about 7cm long and 1.5 cm wide.
2. Label two test tubes (1 & 2) and place one potato strip in each tube.
3. Fill tube 1 with water to cover the potato strip and make a mark with a marker on the tube for
the level of the potato strip.
4. Fill tube 2 with 10% NaCl to cover the potato strip and mark the level of the potato strip.
5. After one hour, observe each strip for limpness (loss of water) or stiffness (gain of water).
B. Diffusion through a semisolid

1. Using a Pasteur pipette, create a hole in the middle of an agar Petri dish.
2. Using a tweezer, add few crystals of potassium permanganate (KMNO4) in the center
depression.
3. Record time zero and the final time. Calculate the length of time in hours and minutes.
4. Measure (using a ruler) in mm the movement of color from the center of depression outward
in one direction. …………………………mm.
5. Calculate the speed of diffusion …………………………mm/hr (Divide the number of
millimeters by the number of hours).
6. Record the data speed of diffusion in table 1 below
C. Diffusion through a liquid
1. Add water to a glass petri dish.
2. Place the petri dish over a thin, flat ruler, as far away from your working space as possible
(Keep the water undisturbed at all time).
3. With tweezers, add a crystal of KMNO4 directly to the side of the petri dish. NOTE THE
TIME ZERO.
4. After 1 hour, note the distance the color has moved. Record the final time.
5. Calculate the speed of diffusion. ……………………. mm/hr.
6. Record your data in the speed of diffusion in table 1.
Table 1: Diffusion Measurements
Medium Time Zero Final time Length of Distance Speed of

time (hrs) moved (mm) diffusion
(mm/hr)
Semisolid
Liquid
D. Effect of tonicity on Red blood cells (RBC) and Print visibility

1. Label 3 test tubes with marker at the top of the tube.
2. To test tube 1 add 2 ml of 0.9% NaCl, to test tube 2 add 2 ml of 20% NaCl and to the final
tube 2 ml of dH2O.
3. Add 3 drops of RBCs to all tubes.
4. Cover with parafilm and then swirl.
5. Leave it to stand for few seconds and then record the tonicity of each in the below table, in
relation to RBCs.
Table 2: Tonicity observations on RBC
Concentration (NaCl) Tonicity Effect on Cells Explanation
0.9% NaCl
20% NaCl
d. H2O (lower than 0.9%
NaCl)
4. Shake each of the above tubes very well and place in front of your lab manual. Determine
whether you can see the print on the page and record your observations in the below table.
Table 3: Print visibility Observations
Tube Tonicity Print Visibility Explanation

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Experiment 6 - Membrane Structure and Function

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Experiment 6 - Membrane Structure and Function

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Membrane Structure and Function

B. Diffusion through a semisolid

Table 1: Diffusion Measurements

Medium Time Zero Final time Length of Distance Speed of

D. Effect of tonicity on Red blood cells (RBC) and Print visibility

Table 3: Print visibility Observations

Tube Tonicity Print Visibility Explanation

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