Unit 8: Review I.

Introduction
Osmosis and Diffusion Lab
Osmosis: the diffusion of water across a selectively permeable membrane from a region
of higher water concentration to a region of lower water concentration.

Osmotic situations:
hypotonic solution – contains less solute compared to another solution
hypertonic solution – contains more solute compared to another solution
isotonic solution – contains the same concentration of solutes compared to another
solution

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II. Effect of temperature Effect of temperature continued

Three agar plates were set up at three temperatures

Diffusion always occurs down the concentration gradient until dynamic equilibrium is 1. room temperature (25°C),
reached. Several factors can affect the rate the diffusion. One such factor is
2. incubation temperature (37°C) and
temperature. Higher temperatures tend to increase the rate of diffusion. Another
factor that affects diffusion is the molecular weight of the diffusing solute. Solutes of 3. refrigeration temperature (5°C).
higher molecular weights tend to diffuse slower than solutes of lower molecular
weights. To each agar plate a drop of potassium permanganate and a drop of Janus Green
Violet color Orange color were added.
Rates of diffusion for two types of molecules were
examined, potassium permanganate (294 Diffusion of dye was measured at 0, 15
Daltons) and Janus Green (511 Daltons) dyes. and 30 minutes of each agar plate.

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III. Effect of solute concentration Effect of solute concentration continued

Five bags were prepared according the information on the table and their weights
measured. The bags were placed into the appropriate solution and left for 60
minutes.
Dialysis tubing is a semi permeable material that can be used to simulate the # Bag contents Beaker contents Osmotic situation
movement of substances through cell membranes (cell membranes are selectively 1 10 ml Distilled water 200 ml Distilled water Isotonic solution
permeable). The pores on the dialysis tubing are large enough to allow water
molecules and other small molecules and ions to move through unrestricted. Larger 2 10 ml 10% sucrose 200 ml Distilled water Hypotonic solution
molecules, such as sucrose, cannot cross because of their size.
3 10 ml 20% sucrose 200 ml Distilled water Hypotonic solution

4 10 ml 40% sucrose 200 ml Distilled water Hypotonic solution

5 10 ml Distilled water 200 ml 40% sucrose Hypertonic solution

Roll of dialysis tubing

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 IV. Differential permeability of membranes Differential permeability of membranes continued

Dialysis tubing is a material that will be used to simulate differentially permeability of
membranes. Only molecules and ions smaller than the pores of the tubing will be
allowed to cross.
The solution contains a mixture of starch, protein, sodium chloride, and sodium sulfate
in an aqueous solution. Starch and protein are both macromolecules. Starch is a
polymer of sugar. Protein is a polymer of amino acids. Sodium chloride (NaCl), salt,
will dissociate in an aqueous solution to produce sodium ion (Na+) and chloride ion
(Cl -). Sodium sulfate (Na2SO4) will dissociate in an aqueous solution to produce
sodium ion (Na+) and sulfate ion (SO42-).
The flask contains starch, protein, sodium chloride, and
sodium sulfate in an aqueous solution. A bag is prepared
containing this cocktail. The bag is then placed in a
beaker of distilled water for 60 minutes. At the end of 60
minutes the water of beaker is tested for the presence of
starch, protein, chloride ions and sulfate ions.

Solution of starch,
protein, sodium
Continue Continue chloride, and sodium
sulfate

Differential permeability of membranes continued Differential permeability of membranes continued

Sample Positive Negative Let’s review the biuret test for protein. If Sample Positive Negative
Let’s review the iodine test for starch. If protein is present in the sample and you
starch is present in the sample and you before starch starch
add biuret, you will see a violet color
before protein protein
add iodine, you will see the solution test test test change. If protein is absent then you see test test test
turn blue-black. If starch is absent then the color of biuret only.
you see the color of iodine only.

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Differential permeability of membranes continued Differential permeability of membranes continued

Silver nitrate (AgNO3) test for chloride ions (Cl-). Sample Positive Negative Barium chloride (BaCl2) test for sulfate ions (SO42-). Sample Positive Negative
The presence of chloride ions are tested with before chloride chloride The presence of sulfate ions are tested with before sulfate sulfate
silver nitrate. If chloride ions are present in barium chloride. If sulfate ions are present in
test ion test ion test test ion test ion test
the sample and you add silver nitrate , you the sample and you add barium chloride ,
will see a milky white precipitate (silver you will see a white precipitate (barium
chloride). If chloride ions are absent then sulfate). If sulfate ions are absent then you will
you will see a clear test tube. Silver nitrate is see a clear test tube. Barium chloride is a
a clear solution. clear solution.
It is difficult to see the white precipitate in this
picture. You have something resembling chalk
dust collecting at the bottom of the test tube.

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 Differential permeability of membranes continued V. Diffusion and osmosis in living cells

These are the results of the tests.

Water flows across membranes unrestricted. It flows from a region of lower solute
concentration to a region of higher solute concentration until equilibrium is
After 60 minutes, samples of the water of the beaker were placed into 4 test tubes. established.

Reagents were added to the test tubes.
Tube 1 received 10 drops of iodine.
Tube 2 received 10 drops of biuret.
Tube 3 received 10 drops of silver nitrate.
Tube 4 received 10 drops of barium chloride.
1 2
In hypotonic solutions, if a cell’s membrane has no support, the cell may rupture
because of an influx of water.
You examined pictures of animal cells (sheep red blood cells) and plant cells
(Elodea leaf cells). Plant cells have cell walls which support the cell and keep
it from osmotic shock. Animal cells do not have cell walls and may rupture
unless they have a mechanism for stabilizing the osmotic differences.
3 4

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Diffusion and osmosis in living cells continued Diffusion and osmosis in living cells continued

Wet mounts of sheep blood

These test tubes contain 4 drops of sheep blood in a Two wet mounts were prepared
saline solution. from the two different blood
– Tube A contains 10% NaCl solution (hypertonic solutions (0.9% NaCl solution
solution). and 10% NaCl solution).
– Tube B contains 0.9% NaCl solution (tonicity 0.9% NaCl solution
similar to that of blood plasma, isotonic A wet mount with 0.45% NaCl
solution). solution was not prepared
– Tube C contains 0.45% NaCl solution (hypotonic because hemolysis occurs at
solution). A B C that concentration and only
cellular debris is seen.
Observe the appearance of each test tube (in the two 10% NaCl solution
pictures). When hemolysis takes place, print behind
the tube can be read. If the cells are intact, then
print behind a tube cannot be read. A label with
print has been placed behind each tube. Can you
see print through the liquid?
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10% 0.9% 0.45%

Diffusion and osmosis in living cells continued Diffusion and osmosis in living cells continued

• Blood slide with 0.9% • Blood slide with 10%

NaCl solution. NaCl solution.
• Normally, blood cells • Blood cells appear
have biconcave crenated (shriveled).
disk shape but • These are cells in a
appear round in 2-D. hypertonic solution.
• These are normal More water moves
cells in an isotonic out of the cells.
solution. Water
moves equally into
and out of the cells,
no net movement of
water.

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Diffusion and osmosis in living cells continued Diffusion and osmosis in living cells continued

• Elodea in 0% NaCl
solution.
Two beakers have been prepared and Elodea
has been placed in each of the beakers. • Cells appear normal
(turgid).
– Beaker A contains 0% NaCl solution
(hypotonic solution). • These are cells in a
hypotonic solution.
– Beaker B contains 20% NaCl solution
More water moves
(hypertonic solution).
into the cells. Notice
how the chloroplasts
Two wet mounts have been prepared from the A B are pushed to the
contents of the two beakers. periphery of the cell
because of the
enlarged central
vacuole.

Beaker A: 0% NaCl
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Beaker B: 20% NaCl

Diffusion and osmosis in living cells continued

• Elodea in20% NaCl

solution.
• Cells appear
plasmolyzed.
• These are cells in a
hypertonic solution.
More water moves
out the cells. Notice
how the chloroplasts
are pushed to the
center of the cell
because of the
collapsed central
vacuole.

End of Lab
Review ☺