INTRODUCTION:

Diffusion and osmosis are passive transport that transports molecules through the plasma
membrane. Diffusion is the movement of molecules or ions from region of high concentration to
region of low concentration down the concentration gradient through the selective permeable
membrane. Only small molecules can be transport by simple diffusion through the selective
permeable membrane. Osmosis is the passage of water molecules from region of high
concentration of water molecules to the region of low concentration of water molecules through
the selective permeable membrane until dynamic equilibrium is achieved. When dynamic
equilibrium is achieved, there will be net movement of molecules or ions because the
concentration gradients are the same. If this occurs, the solution is called isotonic. Meanwhile,
the solution that have high concentration of solute is called hypertonic solution while if the
solution have low concentration of solute, it is called hypotonic solution. Water molecules will
move from the solution that has low concentration of solutes to the solution that have high
concentration of solutes. This is because the water potential of hypotonic solution is higher that
hypertonic solution. Water potential is the measure of potential energy in every water molecules
in a solution. Symbol of water potential is a Greek letter psi (Ψ). Water potential has two
components which are; physical pressure component or pressure potential that symbolize by Ψ P
and the other component is effects of solute or solute potential that represent by the symbol Ψ S.
A formula that use to represent the relationship between these two components is Ψ = Ψ P + ΨS.
Plasmolysis is the effect that occurs when a cell is immersed in the hypertonic solution.
Plasmolysis is contraction of the cytoplasm of a plant cell as a result of loss of water from the
cell to the hypertonic solution. The plant cell will undergo deplasmolysis when it is immersed in
hypotonic solution. If animal cell is immersed in hypertonic solution, the cell will be crenelated
while if it is immersed in the hypotonic solution, the cell will undergo hemolysis and eventually
will burst.
OBJECTIVES:

Before doing this lab I should understand:

 The mechanisms of diffusion and osmosis and their importance to cells;

 The effects of solute size and concentration gradients on diffusion across selective
permeable membrane;
 The effects of a selective permeable membrane on diffusion and osmosis between two
solutions separated by the membrane;
 The concept of water potential;
 The relationship between solute concentration and pressure potential and the water
potential of the solution; and
 The concept of molarity and its relationship to osmotic concentration.

After doing this lab I should be able to:

 Measure the water potential of a solution in a controlled experiment;

 Determine the osmotic concentration of living tissue or an unknown solution from
experimental data;
 Describe the effects of water gain or loss in animal and plant cells; and
 Relate osmotic potential to solute concentration and water potential.

EXERCISE 1A : DIFFUSION

HYPOTHESIS:

Only small molecules that are water molecules and glucose molecules can pass through the
selective permeable membrane that is dialysis tubing, while the large molecules which are starch
molecules cannot pass through the dialysis tubing.

MATERIALS AND APPARATUS:

20-cm piece of 2.5-cm dialysis tubing, 15% glucose solution, 1% starch solution, Benedict’s
solution or glucose Testape(r) or Clinistirx(r) solution, distilled water, Lugol’s solution, 250-mL
beaker, and measuring cylinder.
PROCEDURE:

1. A 20-cm piece of 2.5-cm dialysis tubing that has been soaking in the water is obtained.
One end of the tubing is tied off to form a bag. The other end of the bag is opened by
rubbing the end of the tubing between the fingers until the edges separate.
2. The 15% glucose + 1% starch solution is tested for the presence of glucose by doing
either Benedict’s test or glucose Testape(r) or Clinistix(r). The results are recorded.
3. 15 mL of the 15% glucose/1% starch solution is placed in the bag. The other end of the
bag is tied off, leaving sufficient space for expansion of the content in the bag. The color
of the solution is recorded.
4. A 250-mL beaker is filled with distilled water. 4 mL of Lugol’s solution is added
approximately to the distilled water and the color of the solution is recorded. This
solution is tested for glucose and the result obtained is recorded.
5. The bag is immersed in the beaker of solution.
6. The setup is leaved for approximately 30 minutes or a distinct colour change is seen in
the bag or in the beaker. The final colour of the solution in the bag and of the the solution
in the beaker are recorded.
7. The liquid in the beaker and in the bag are tested for the presence of glucose. The results
obtained are recorded.

EXERCISE 1B : OSMOSIS

HYPOTHESIS:

The net movement of water molecules during osmosis is from hypotonic solution that has lower
concentration of solute and higher water potential to the hypertonic solution that has higher
concentration of solute and lower water potential.

MATERIALS AND APPARATUS:

30-cm strips of dialysis tubing, distilled water, 0.2 M sucrose solution, 0.4 M sucrose solution,
0.6 M sucrose solution, 0.8 M sucrose solution, 1.0 M sucrose solution, paper towels, 250-mL
beakers, electronic balance.
PROCEDURE:

1. Six 30-cm strips of pre-soaked dialysis tubing are obtained.

2. A knot is tied in one end of each piece of dialysis tubing to form 6 bags. 15-25 mL of the
following solutions were poured approximately into separate bags:
a. Distilled water
b. 0.2 M sucrose
c. 0.4 M sucrose
d. 0.6 M sucrose
e. 0.8 M sucrose
f. 1.0 M sucrose
Most of the air from each bag is removed by drawing the dialysis bag between two
fingers. The other end of the bag is tied off. Sufficient space is leaved for expansion of
the contents in the bag. The solution only filled about one-third to one-half of the piece of
tubing
3. Each bag is rinse gently with distilled water to remove any sucrose spilled during the
filling.
4. The outside of each bag is blotted carefully and the initial mass of each bag that has been
expressed in grams is recorded.
5. Each bag is placed in an empty 250-mL beaker and the beaker is labeled to indicate the
molarity of the solution in the dialysis bag.
6. Each beaker is filled two-thirds full with distilled water. Each bag is sure to completely
submerge.
7. The setup is leaved for 30 minutes
8. At the end of 30 minutes, the bags are removed from the water. The bags are blotted
carefully and the final mass of each bag is determined.
9. The results obtained are recorded.
EXERCISE 1C : WATER POTENTIAL

HYPOTHESIS:

As the molarity of sucrose solution increase, the weight of the potato cylinders immersed in the
sucrose solution will decrease.

MATERIALS AND APPARATUS:

Potatoes, 0.2 M sucrose solution, 0.4 M sucrose solution, 0.6 M sucrose solution, 0.8 M sucrose
solution, 1.0 M sucrose solution, distilled water, 250-mL beakers, cork borer, electronic balance,
plastic wraps, paper towels, and ruler.

PROCEDURES:

1. 100 mL of assigned solution is poured into a labeled 350-mL beaker. A potato is

sliced into discs that are approximately 3 cm thick.
2. A approximately 5 mm of inner diameter of cork borer is used to cut four potato
cylinders. The potato’s skin is not included on the cylinders. Four potato cylinders are
needed for each beaker.
3. The potato cylinders are kept in a covered beaker until it is turn to use the balance.
4. The mass of four cylinders were determined together and the mass is recorded. The
four cylinders are put into each beaker containing sucrose solution of 0.2 M, 0.4 M,
0.6 M, 0.8 M and 1.0 M sucrose solution. Another four cylinders are put into a beaker
of distilled water.
5. The beaker is covered with plastic wrap to prevent evaporation. Each beaker is
labeled.
6. The setup is leaved overnight
7. The cores are removed from the beakers, blotted them gently on paper towels, and
their total mass is determined.
8. The final mass is recorded and the percentage change is calculated.
9. The data of percentage change in mass is graphed.
EXERCISE 1D : CALCULATION OF WATER POTENTIAL FROM SOLUTE
POTENTIAL OF A SOLUTION

PROCEDURES:

1. The solute potential of the sucrose solution is calculated using the formula:
ΨS = - iCTR
Where:
i = Ionisation constant (0.1)
C = Molar concentration (of sucrose solution)
T = Temperature °K (273 + °C of solution)
R = Pressure constant (R = 0.0813 litre bars/mole °K)

2. Water potential, Ψ is calculated. The water potential will be equal to the solute
potential of the solution. (ΨP = 0)
Ψ = 0 + ΨS or Ψ = ΨS
3. The water potential, Ψ of potato cells is determined.

EXERCISE 1E : ONION CELL PLASMOLYSIS

HYPOTHESIS:

When the salt solution is added, the onion epidermis cells will experience plasmolysis while
when the fresh water is added, deplasmolysis of the cells will occur.

MATERIALS AND APPARATUS:

Epidermis of onion, 15% sodium chloride, NaCl, fresh water, microscope, cover slip and slide,
and paper towel.
PROCEDURE:

1. A wet mount of a small piece of epidermis of an onion is prepared. The epidermis

is observed under 100X magnification. The appearance of the onion cells are
sketched and described.
2. 2 or 3 drops of 15% NaCl are added to one edge of the cover slip. This salt
solution is drew across the slide by touching a piece of paper towel to the fluid
under the opposite edge of the cover slip. The onion cells are sketched and
described. The observation is explained.
3. The cover slip is removed and the onion epidermis is flooded with fresh water.
The epidermis was observed under 100X magnification of microscope. The
observation is described and explained.
ERROR ANALYSIS

EXERCISE 1A:

The data collected in this experiment agreed with the theory learned. Thus, no error
detected in the experiment.

EXERCISE 1B:

In this experiment, the error that we made was when we tied the dialysis tubing. We did
not tie the dialysis tubing tight enough especially the 0.0 distilled water and 1.0M sucrose
solution dialysis tubing. As a result, the contents of the dialysis tubing bags spilt out from the
bag and affected the final mass of the bag.

EXERCISE 1C:

The data collected from the experiment does not show any error and agreed with the
theory learned.

EXERCISE 1D:

In this experiment, only simple math calculation is made. Thus, possible error of the
experiment is mathematical error such as algebraic formula and decimals.

EXERCISE 1E

In this experiment, error that I made was not preparing the wet sample correctly causing
difficulties to observe the onion cell under the microscope. There also possibility that the onion
cells dried out because it was exposed to the air for a long time. Thus, no reaction occurs when
15% of sodium chloride is added to the onion cells.
DISCUSSION

EXERCISE 1A:

Diffusion is the movement of molecules or ions from region of high concentration to

region of low concentration down the concentration gradient through the selective permeable
membrane. The diffusion of the molecules also based on the size of the molecules to pass
through the membrane pore. Glucose molecules, Iodine Potassium Iodide (IKI) and water
molecules has smaller relative size, while starch molecules have bigger relative size as compared
to the membrane pore of dialysis tubing bag. Based on the data collected from experiment, the
glucose molecules and iodine potassium iodide (IKI) molecules diffuse down the concentration
gradient. The glucose molecules diffuse out from the dialysis tubing bag that has higher
concentration of glucose molecules to the beaker that has lower concentration of glucose
molecules. IKI molecules on th other hand diffuse from the beaker that has higher concentration
of IKI molecules into the dialysis tubing bag that has lower concentration of IKI molecules.
While starch molecules stayed in the dialysis tubing bag. This was proved when we test the
solution in the beaker with Benedict’s test and we got brick red precipitate that shows the present
of glucose in the beaker. The colour change of the solution in the beaker and the dialysis tubing
bag proved that the concentration of IKI molecules is decreasing in the beaker and there is
presence of starch molecules and IKI molecules in the dialysis tubing bag because both of the
molecules react with each other resulting blue black solution in the dialysis tubing bag.

EXERCISE 1B:

Osmosis is the passage of water molecules from region of high concentration of water
molecules to the region of low concentration of water molecules through the selective permeable
membrane until dynamic equilibrium is achieved. When the solution in a dialysis tubing bag has
higher concentration of solute (lower concentration of water molecules) than the surrounding, the
solution is hypertonic to the surrounding. Thus, water molecules from surrounding that has
higher concentration of water molecules will diffuse into the bag until dynamic equilibrium is
achieved. When the solution in a dialysis tubing bag has lower concentration of solute (higher
concentration of water molecules) than the surrounding, the solution is hypotonic to the
surrounding. Thus, water molecules from surrounding that has higher concentration of water
molecules will diffuse into the bag until no net movement of water molecules between the
surrounding and the bag. When there is no net movement of water molecules, the solution is
isotonic. Based on the experiment, when the molarity of the sucrose solution in the dialysis
tubing bag increases, the percent change in mass of the dialysis tubing bag increases. Thus, the
sucrose solution is hypertonic to the distilled water in the beaker. When the molarity of sucrose
solution increases, the amount of sucrose molecules in the solution increases. Thus, more water
molecules will diffuse into the dialysis tubing bag due to osmosis. So, the mass change of the
bag increases.

EXERCISE 1C:

Water potential is the measure of potential energy in every water molecules in a solution.
When there is higher water molecules in a solution compared to solute molecules, the solution
has high water potential. While when there is lower water molecules in a solution compared to
solute molecules, the solution has low water potential. Based on the experiment, the water
potential of distilled water and 0.2M of sucrose solution is higher than the water potential of
potato cores. Water molecules from distilled water and 0.2M of sucrose solution diffuse into the
potato cores due to the osmosis. Thus, the change in mass of the potato cores is positive. The
water potential of 0.4M, 0.6M, 0.8M, and 1.0M of sucrose solution is lower than the water
potential of potato cores. Water molecules from 0.4M, 0.6M, 0.8M, and 1.0M of sucrose solution
diffuse out from the potato cores due to the osmosis. Thus, the change in mass of the potato cores
is negative. Based on the graph that I drew with line of best fit, I predicted that 0.38M of sucrose
solution is isotonic to the potato cores because the percent change of mass of potato cores is zero.

EXERCISE 1D:

In this experiment, all these conclusion of the previous experiments were reinforced with
scientific equations. The calculation made is to prove that all the results are correct based on the
theories learned. The calculation of water potential using formula; Ψ = ΨS and ΨS = - iCTR,
helped to give better understanding about solute and water potential.
EXERCISE 1E:

Plasmolysis is the effect that occurs when a cell is immersed in the hypertonic solution.
In this experiment, the onion cells was tested and observed under the microscope. When the 15%
of sodium chloride (NaCl) was added to the onion cells, the vacuole of the cells shrunk and the
cytoplasm is pulled away from the cell wall. This reaction is plasmolysis. It happened because
15% of NaCl is hypertonic to the onion cell. Thus, the water molecules from the onion cells that
have higher water potential diffuse out to the surrounding that has lower water potential due to
osmosis. When the onion cells were flooded with fresh water, the vacuole of the cells expanded
and pushed the cytoplasm against the cell wall. This reaction is called deplasmolysis. It happened
because fresh water is hypotonic to the onion cell. Therefore, the water molecules from the fresh
water that has higher water potential diffuse into the onion cells that have lower water potential
via osmosis. The cell wall of the onion cells prevents the cells from bursting.
CONCLUSION

EXERCISE 1A:

The result of the experiment conclude that glucose molecules, Iodine Potassium Iodide
(IKI) and water molecules has smaller relative size, while starch molecules have bigger relative
size as compared to the membrane pore of dialysis tubing bag. This proved that the hypothesis is
correct. Only small molecules that are water molecules and glucose molecules can pass through
the selective permeable membrane that is dialysis tubing, while the large molecules which are
starch molecules cannot pass through the dialysis tubing.

EXERCISE 1B:

The result of the experiment is when the molarity of the sucrose solution in the dialysis
tubing bag increases, the percent change in mass of the dialysis tubing bag increases. Thus, the
sucrose solution is hypertonic to the distilled water in the beaker. Therefore, water molecules
diffuse from distilled water into the dialysis tubing bag. This proved that the hypothesis is
correct. The net movement of water molecules during osmosis is from hypotonic solution that
has lower concentration of solute and higher water potential to the hypertonic solution that has
higher concentration of solute and lower water potential.

EXERCISE 1C:

The hypothesis of the experiment is as the molarity of sucrose solution increase, the
weight of the potato cylinders immersed in the sucrose solution will decrease. Based on the
experiment, the hypothesis stated is not accurate. The result of the experiments is when potato
cores are immersed in distilled water and 0.2M of sucrose solution, the percentage change in
mass of the potato cores is positive. When potato cores are immersed in 0.4M, 0.6M, 0.8M, and
1.0M of sucrose solution, the percentage change in mass of the potato cores is negative. Thus,
the conclusion of the experiment is when the potato cores is immersed in the hypotonic solution,
the final mass of the potato cores increases while when the potato cores are immersed in
hypertonic solution, the final mass of the potato cores decreases.
EXERCISE 1D:

This part of exercise proved that the data collected throughout the experiments were
correct through the calculation using the given formula. The calculation water and solute
potential give the better understanding on the concept of diffusion of water molecules or
osmosis.

EXERCISE 1E:

The hypothesis of the experiment is when the salt solution is added, the onion epidermis
cells will experience plasmolysis while when the fresh water is added, deplasmolysis of the cells
will occur. In the experiment, when the 15% of sodium chloride (NaCl) was added to the onion
cells, the vacuole of the cells shrunk and the cytoplasm is pulled away from the cell wall while
when the onion cells were flooded with fresh water, the vacuole of the cells expanded and
pushed the cytoplasm against the cell wall. The hypothesis stated is accepted.
REFERENCE

 Osmosis (s.a). on Wikipedia: https://en.wikipedia.org/wiki/Osmosis

 Diffusion (s.a). on Wikipedia: https://en.wikipedia.org/wiki/Diffusion
 Jane B Reece, Neil A Campbell. Campbell Biology (9th ed). Boston : Benjamin
Cummings. Pearson.