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Diffusion

and
Osmosis
Lab
AP Biology Lab #1

Janice Chan

Period 2

Partners: Guneet
Kaur, Kevin Ta
Janice Chan

Dr. Fuller
AP Biology – Period 2
30 September 2010
AP Biology Lab #1 – Diffusion and Osmosis
I. Pre-lab Questions
1. What are the mechanisms of diffusion and osmosis and their importance to
cells?
 Diffusion is known as the movement of molecules from a higher concentration to
a lower concentration. Osmosis is a special type of diffusion in which the action
occurs, “in water separated by a selectively permeable membrane, with different
solute concentrations on either side of the membrane… In osmosis, the water
moves from a region of low solute concentration to a region of high solute
concentration.” The mechanisms are important to cells because they allow for
many life functions of the cell, including the transportation of “vitally important
nutrients and compounds without expenditure of excess metabolic energy.”
Osmosis and its impact on plant cells allows for, “the absorption and early
transport of water into the root system of plants, and, with transpirational pull,
helps transport water in the xylem.
2. What are the effects of solute size and concentration gradients on diffusion and
osmosis between two solutions separated by the membrane?
 The solute size does not affect the diffusion of the membrane, but the
concentration gradient does. The higher the concentration, the faster the water
will flow through the membrane.
3. What are the effects of a selectively permeable membrane on diffusion and
osmosis between two solutions separated by the membrane?
 When you have a membrane that separates two solutions, it restricts certain
materials from passing freely through the membrane. With diffusion and osmosis,
the membrane allows certain, “small, neutrally charged molecules, such as
oxygen, carbon dioxide, water, and glucose to pass freely.”
 The selectively permeable membrane has different cellular concentrations on
either side, so the osmosis moves the solution from lower concentration to higher
concentration.
4. What is water potential?
 A solute’s relative concentration; Water always moves from an area of higher
water potential to an area of lower water potential.
5. What is the relationship between solute concentration, pressure potential, and
the water potential of a solution?
 Pressure potential and osmotic potential assist water potential in reaching
equilibrium.
 Solute concentration equals water potential, and when the pressure potential is
equal to the osmotic potential, water potential is 0, and equilibrium has been
reached.
6. What is the relationship of molarity to osmotic concentration?
 As molarity increases, so does osmotic concentration.

Diffusion and Osmosis Lab
I. Title
a. Diffusion and Osmosis

II. Objective
a. To investigate the processes of diffusion and osmosis in a model of a membrane
system.
b. To investigate the effect of solute concentration on water potential as it relates to
living plant tissues.

III. Hypothesis
a. The starch will diffuse, turning the solution purple. The glucose will remain in the
bag.
i. Starch can’t go through, but glucose can, meaning that the starch did not
diffuse and my hypothesis was incorrect.
b. The mass will increase as the concentration increases.
i. The mass did increase, proving diffusion into the bag.
c. .6 will match the osmotic molar concentration of the potato the most.
i. .4 M matched the osmotic molar concentration of the potato.
d. The onion skin will dissolve the salt solution. [The salt will dissolve into the
onion.]
i. The onion’s nuclei were more visible once the NaCl had absorbed into its
skin.

IV. Materials
1. 4 glucose indicator strips.
2. 1 graduated cylinder.
3. 1 plastic cup, 250 ml dialysis tubing, 7 ft.
4. 6 plastic cups, 250 ml.
5. Glucose starch solution, 15 ml.
6. IKI [potassium iodide] solution, 1ml.
7. Sucrose solution, .2 M, 175 ml.
8. Sucrose solution, .4 M, 175 ml.
9. Sucrose solution, .6 M, 175 ml.
10. Sucrose solution, .8 M, 175 ml.
11. Sucrose solution, 1.0 M, 175 ml.
12. Distilled water, 175 ml.
13. Paper towels.
14. Potato.
15. Plastic wrap.
16. Microscope slide.
17. Cover slip.
18. Compound microscope.
19. Scalpel.
20. Forceps.
21. Balance, compound microscope, cork borer with plunger, NaCl solution, 15%,
Onion epidermis.
V. Procedure
a. Part A - Diffusion
i. Pour 15 ml of prepared glucose/starch solution into graduated cylinder.
ii. Obtain piece of dialysis tubing, soaked in water, tie TIGHT knot in one
end of tubing.
iii. Open tubing by rubbing between fingers.
1. Pour 15 ml of solution [glucose/starch] into tubing.
iv. Note the color of solution in bag, record color.
v. Determine if glucose is present by dipping glucose indicator strip into
solution. Record data.
vi. Tie knot in open end to form a bag, leave enough space in bag for
expansion.
vii. Fill 250 ml beaker or plastic cup 2/3 full with distilled water. Add 1 ml of
potassium iodide [IKI] to beaker.
viii. Note color, record color.
ix. Determine glucose presence; dip another indicator strip, record data.
x. Completely immerse dialysis bag into solution in the beaker.
xi. Wait 30 minutes.
xii. Remove dialysis bag from beaker. Record final color.
xiii. Determine glucose content, to test solution, make small cut in bag with
scissors, and insert strip through the hole. Record presence / absence of
glucose in Table.
b. Part B - Osmosis
i. Obtain six plastic cups, label them as follows: water, .2M, .4M, .6M, .8M,
1.0M.
ii. Obtain six pieces of dialysis tubing from beaker of water, Tie tight knot in
one end of tubing.
iii. Open one piece of tubing, rub between fingers to open. Pour 25 ml
distilled water into tubing, carefully tie knot in open end to form a bag.
Leave enough space for expansion, blot dry, and place in cup labeled
“water.”
iv. Repeat procedure with remaining pieces of dialysis tubing, add different
sucrose solution to each bag: .2, .4, .6, .8, 1.0M.
v. Blot bags dry, weigh each one. Record initial mass in data table 2.
vi. Fill the six plastic cups with approximately ¾ distilled water in each one.
Immerse one bag in each cup, be sure each bag is the properly labeled cup.
vii. Wait 30 minutes. Remove bag from cups, Blot dry, weigh each one again.
viii. Record final mass in table2.
ix. Calculate percent change in mass for each bag. Formula: %Change=
[Final Mass – Intial Mass/Initial Mass x100.]
x. Graph both individual and class average on the graph paper in Analysis
section.
c. Part C – Water Potential
i. With cork borer, cut four cylinders from potato for each solution you will
be using. [Our group, Water, and .4M] Cut each cylinder to a length of
3CM for greater accuracy; remove any skin from the cylinders, place
cylinders in a covered cup/beaker.
ii. Weigh the four cylinders together, record initial mass in Table 3.
iii. Pour 100 ml of solution you have been assigned into beaker/plastic cup.
Take initial temps and record in Table 3. Insert four potato cylinders and
cover beaker or cup with plastic wrap.
iv. Repeat procedure for other solutions, let beakers stand overnight.
v. Next day, measure final temp of liquid in beakers. Record temp in Table 3.
vi. Remove cylinders from beaker, blot carefully with paper towel and weigh
them. Record final mass in Table 3.
vii. Calculate percent change in mass for four potato cylinders using same
formula as above.
viii. Record data in Table 3.
ix. Graph both individual and class average results, place percent change in
mass on y-axis and sucrose molarity on x-axis. Using graph, determine
molar concentration of the potato cylinders. This is equivalent to sucrose
molarity in which potato core mass is constant.
d. Part D – Water Potential Calculation
i. Osmotic potential calculated using: -1 [for sucrose] x .4M x .0831 x 296K
ii. -1 x .4 x .0831 x 296 = = -9.83904 Water potential
e. Part E – Plant Cell Plasmolysis
i. Cut an onion in half lengthwise, from top to bottom, remove center. Note
layers of the scale leaves in onion. The epidermal layer, only one cell
thick, between each scale is the layer used in the experiment.
ii. Make several shallow cuts approx. 1 cm apart on one of the scale leaves.
Make more cuts perpendicular to first set. Results in 1x1cm squares.
iii. Using forceps, carefully remove epidermal layer from squares. If layer
tears, take another layer from other squares.
iv. Place layer in two to three drops of distilled water on microscope slide.
Place coverslip on top.
v. Examine cell layer under 100X magnification and note characteristics of
cells. Draw cells in Analysis section.
vi. Remove cell from microscope, add two to three drops of 15% NaCl
solution to one side of slip.
vii. Carefully touch edge of paper towel to other side in order to draw NaCl
across the sample.
viii. Allow the slide to sit for two to three minutes in salt solution, re-examine
the sample under the microscope.
ix. Note appearance of cells, Draw in Analysis section.
VI. Data
VII. Post-Lab Questions
1. In Part A, what molecules could diffuse through the dialysis tubing and
which stayed inside? Why?
 The glucose was able to diffuse through the tubing because of its smaller
molecular structure and its lower density.
2. In Part B, why did the mass of the bags change?
 The mass of the bags changed because of the bag’s higher concentration. The
solution naturally moved from the water in the cup to the bag due to osmosis,
which adds mass to the bag.
3. If the bags in Part B were placed in .4 M sucrose instead of distilled water,
what would the results be?
 The results would change, because everything below .4 M would decrease in
mass, while the bags above .4 M would increase in mass. The .4 M bag would not
change at all, because it is at equilibrium with the solution.
4. Why is measuring the temperature an important part of the calculation of
osmotic potential?
 Without the temperature, the potential could be too varied for the experiment. The
heat or cold would change the data.
5. If the potato cores in Part C were allowed to dry out before use in this
experiment, how would the results be affected?
 They would gain more water if they were allowed to dry out before use. More
water would be absorbed when you placed the potato core into the water solution.

VIII. Conclusion
This lab was conducted in order to show us the process of diffusion and osmosis up
close. We first started with Diffusion, in which we took a dialysis tubing and tied off
its ends, filled with starch and glucose, then placed it in a water solution. Because the
glucose molecules were smaller and less dense than the starch ones, the starch
remained in the bag, while the glucose molecules diffused out of the bag, blending
with the distilled water in the beaker and giving our indicator a positive reading on
glucose presence. In Part B of our experiment, we watched Osmosis by taking more
dialysis bags and tying them off, filled with a given amount of sucrose solution. After
we had tied them off, filled with their either, .2, .4, .6, .8, or 1.0M sucrose solution,
we put them into ¾ full cups of distilled water. We waited to see if the weight in the
bags would change due to osmosis, because the water had a lower concentration
compared to the bag, naturally, molecules within the distilled water would transfer
into the bag, adding weight. In Part C, we found the osmotic potential of the potato
cylinders, which we then graphed. Through the graphs, we found that .4M matched
the concentration the most. In Part D, we calculated the total Water Potential, which
came to -9.83904. In Part E, we found that once the onion cell had absorbed the NaCl,
its nuclei became more visible through the microscope. It has shown me through
experiment how diffusion and osmosis works, and the relationship between the
concepts, including water potential, pressure potential, as well as osmotic molarity. It
has shown how crucial it is for a cell to possess these qualities, in order to function
and operate efficiently. My experiment supported half of my hypotheses. In Part A,
my hypothesis was incorrect, I supposed that because the starch was a solution as
well, it would diffuse out of the bag, despite its higher density and molecular size. I
was wrong, the glucose diffused much more quickly and blended with the water in
the beaker due to its smaller molecules and lower density, which also demonstrated
the law of diffusion. My hypothesis for Part B was correct, in osmosis, the solution is
meant to move to an area of higher concentration. The higher concentration was
within the dialysis bags, because the water had a lower and lower concentration as the
bags’ sucrose solution increased, thus proving that as the mass increased, the
concentration also would. In Part C, I guessed wrongly and believed that .6M would
be the amount of sucrose that matched the osmotic molar concentration of the potato,
while really, it was .4M, I found this out through my graphing, because the x-axis
intersected at the .4M mark. In Part E, I thought that nothing would occur, and the
onion would merely absorb the salt solution. However, when the salt solution was
placed, the onion absorbed it and once put under a microscope, the onion looked
smaller, more shriveled. This is because the salt solution is a hypertonic one; the
concentration of the solute outside of the cell is higher than the inside’s. This means
the concentration of water is smaller in the outside of the cell than the inside, which
makes the water molecules within the cell diffuse from the inside to the outside,
resulting in the cell’s shrunken state. This lab was very thorough in its explaining of
diffusion and osmosis. However, I think that if there had been time to measure the
onion cell a couple times, we would have gotten the point of the experiment across
more clearly. I was quite puzzled at first, before I reread the pre-reading and
understood the process. This lab is a useful teaching tool, while its multi-faceted
procedure can help us understand many other processes about diffusion and osmosis.
Overall, I believe my group was successful in this lab. I corrected my hypotheses and
learned more about the process of diffusion and osmosis both within a cell, and
within molecules.

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