Diffusion & Osmosis Lab

TM
AP Biology Lab 1
Osmosis and Diffusion
Lab Activity
Student Study Guide
BACKGROUND
DID YOU KNOW?

Osmosis is fundamental in
many industrial processes
such as water purification,
desalination, and waste material treatment.
If a bottle of perfume is opened at one end of the room, in a fairly

short time someone at the other end of the room will be able to detect
the scent. How does this happen? It depends on the movement of
molecules. All molecules in gases and liquids are in a state of constant, random motion. The molecules in the perfume, released from
the bottle, travel through the air. When molecules are closely packed
together, they collide with other molecules, their kinetic energy is redirected, and the molecules move away from the point of the collision. As a result, molecules tend to distribute themselves evenly
throughout the area they occupy. When the molecules are evenly distributed, they reach dynamic equilibrium. Individual molecules are
still in motion, but there is no net movement of molecules; as many
molecules move in one direction as they do in the opposite one. This
movement from a higher concentration to a lower concentration is
known as diffusion.
Figure 1
Water
Sugar
Copymaster. Permission granted to make unlimited copies for use in any one
school building. For educational use only. Not for commercial use or resale.
2002 WARDS Natural Science Establishment, Inc.

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Diffusion also occurs in water separated by a selectively permeable

membrane, with different solute concentrations on either side of the
membrane; this special kind of diffusion is known as osmosis. In osmosis, water moves from regions of low solute concentration to regions of high solute concentration. Because systems always tend toward equilibrium, this movement occurs to even out the concentrations on the two sides of the membrane (Figure 1).
Only a solutes relative concentration, or water potential, affects the
rate of osmosis; the size of the solute molecules does not matter. The
higher the concentration of solutes, the faster water will flow through
the membrane to equalize the concentration. This is due to the higher
water potential of the solution.
Isotonic solution:
The solute concentration outside of the cell is the same as
the solute concentration inside the cell.
Hypotonic solution:
The solute concentration outside of the cell is lower than
Hypertonic solution:
The solute concentration outside of the cell is higher than
Osmosis occurs in both plant and animal cells, across a cells plasma
membrane. The membrane allows the free passage of some materials
while restricting the diffusion of others. Small, neutrally charged
molecules such as oxygen, carbon dioxide, water, and glucose can
pass freely through the plasma membrane.
Diffusion is vital to many life functions of a cell, it allows the transport of vitally important nutrients and compounds without the expenditure of excess metabolic energy. Diffusion is also responsible for
the transport and exchange of oxygen and carbon dioxide in the
lungs. During respiratory gas exchange, oxygen is broken down and
carbon dioxide produced. As a result, the concentration of oxygen is
always lower inside the cell than outside, while the concentration of
carbon dioxide is higher inside. The gases travel down their respective concentration gradients through the bodys intercellular fluid.
Oxygen is picked up by the oxygen-poor blood as it passes through
the lungs and diffuses into the body cells as carbon dioxide
diffuses out.
In plants, diffusion aids in the transport of water and nutrients in the
xylem. A plants root cells expend energy to maintain a high solute
concentration in the xylem, which in turn forms a concentration gradient within the roots for water; a higher solute concentration equals
lower water concentration. Water flows down the concentration gradient, causing it to diffuse into the roots. Water pressure builds up
within the xylem, forcing it up the stem. Osmosis 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.
Osmosis also has a major impact on living cells. Organisms rarely exist in environments with solute concentrations that match their cytoplasm; there are usually more or fewer dissolved particles in one of
two compared solutions separated by a membrane, such as a cell and
the media in which it exists. A solution is hypotonic if it has fewer
dissolved particles than the other solution and hypertonic if it has
more particles than the other solution. If the number of dissolved particles are the same in the two media, it is called isotonic.

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Figure 2
Plant Cells
Hypertonic
DID YOU KNOW?

Osmosis was originally discovered by a French physicist,
Abb Jean Antione Nollet,
in 1748.
Isotonic
Hypotonic
If a cell is hypertonic to its surroundings, water rushes into the cell,

causing it to expand. If this occurs in an animal cell, the cell bursts, or
cytolizes. In a plant cell, however, the rigid wall of the cell holds the
cell together. Eventually the pressure exerted by the wall on the water in the cytoplasm will equal the osmotic potential of the solution,
and the net movement of water will become zero. Water will still be
moving, but the amount moving into the cells from osmosis is the
same as the amount being forced out by the cell wall pressure. This
pressure exerted on the cell wall by the water is known as turgor; it is
responsible for supporting the stems and leaves of plants.
If a cell is hypotonic to its surroundings, it loses water, causing it to
shrink. This is called plasmolysis. In plants, the plasma membrane
will pull away from the cell wall; the loss of pressure on the cell wall
is what causes plants to wilt. Animal cells, when plasmolyzed, lose
shape and support, and shrivel up.
Animals in hypertonic media, like saltwater fish, must find ways to
retain their water, while animals in hypotonic media, like paramecia
in freshwater ponds, must develop mechanisms to pump out the excess water they absorb.
Rates of diffusion and osmosis, and the water potential of a solution,
can all be determined through lab activities and calculations.
The osmotic potential is calculated using known concentrations of
solution. A solution of sugar and water is placed in a tube and the
tube is covered by a selectively permeable membrane. If this tube is
placed in a beaker of pure water, the water will diffuse into the tube
and the level of the water will rise. Determining how high the water
rises helps ascertain the water potential of the solution. Water potential determines the direction and rate of osmosis. It consists of two
components: pressure potential, which is the exertion of pressure on a
solution, and osmotic potential, which is the relative concentration of
solutes within the two solutions.

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DID YOU KNOW?

It is because of diffusion that
just one quart of oil from an
oil spill can contaminate up to
250 thousand gallons of
drinking water or cause an oil
slick nearly 2 acres in size.
The water initially enters the tube because there is a negative osmotic
potential in the sugarwater solution. However, the force of gravity
begins to exert pressure on the rising column. When the force of gravity (pressure potential) equals the osmotic potential, the column stops
rising. The water potential is at zero and an equilibrium has been established. The pressure potential can be determined from the height
of the column. With the water potential and the pressure known, osmotic potential can be easily determined.
Water Potential () = Pressure Potential (p) + Osmotic Potential ()
The water potential of a solution is determined by calculating the osmotic potential when the pressure potential is zero.
= p + ; if p = 0, then =

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OBJECTIVES
Demonstrate diffusion across a semi-permeable membrane

Measure the effects of various concentrations of solute on the
process of osmosis
Differentiate between hypotonic, isotonic, and hypertonic environments
Measure and calculate water potential
Examine the effects of osmosis on plant cells
MATERIALS
MATERIALS NEEDED PER GROUP
4
1
1
6
DID YOU KNOW?

Frogs never drink water.
They absorb water from their
environment through the
process of osmosis.
Glucose indicator strips

Graduated cylinder
Plastic cup, 250 ml dialysis tubing, 7 ft.
Plastic cups, 250 ml
Glucose starch solution, 15 ml
IKI (potassium iodide) solution, 1 ml
Sucrose solution, 0.2 M, 175 ml
Distilled water, 175 ml
Paper towels
Potato
Plastic wrap
Microscope slide
Coverslip
Compound microscope
Scalpel
Forceps
SHARED MATERIALS
Balance
Compound microscope
Cork borer with plunger
NaCl solution, 15%
Onion epidermis

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PROCEDURE
As a general laboratory practice, it is recommended that students wear proper protective equipment such as gloves, safety
goggles, and a lab apron to avoid staining any clothing or skin.
A. Diffusion
1. Pour 15 ml of the prepared glucose/starch solution into a graduated cylinder.
NOTE
Solute:
The constituent of a solution
that is dissolved in the solvent.
The pores in the dialysis tubing are extremely small, and

can be easily clogged by any oil or dirt on your fingers
and hands. Wash your hands before handling the dialysis
tubing, and keep physical contact with the tubing to
a minimum.
2. Obtain a piece of dialysis tubing that has been soaking in water.

Tie a tight knot in one end of the tubing, or use a piece of string to
tie off the end.
Solvent:
The constituent of a solution
that dissolves the solute.
NOTE
3.
Concentration:
The ratio of the mass or volume of a solute to the mass or
volume of the solvent.
If you choose to tie off the end of the dialysis tubing with
string, tie two knots, about 1/4 apart, to prevent leaking.
Open the tubing by rubbing the untied end between your fingers.
Pour 15 ml of the glucose/starch solution into the tubing.
4. Note the color of the solution in the bag. Record the color in Table
1 in the Analysis section.
5. Determine if glucose is present in the tubing by dipping a glucose
indicator strip into the solution. Record the data in Table 1.
6. Carefully tie a knot in the open end to form a bag. Be sure to
leave enough space in the bag for expansion.
7. Fill a 250 ml beaker or a provided plastic cup approximately 2/3
full with distilled water. Add 1 ml of potassium iodide (IKI) to
the beaker.
CAUTION
The IKI solution is an irritant; it affects skin and eyes,

and can stain clothing. Handle the solution with caution.
Wash off spills and splashes with water.
8. Note the color of the solution. Record the color in Table 1.

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9. Determine if glucose is present in the beaker by dipping a glucose

indicator strip into the solution in the beaker. Record the data in
Table 1.
10. Completely immerse the dialysis bag in the solution in the beaker.
11. Wait 30 minutes.
NOTE
Though a color change may become apparent after a few

minutes, be sure to allow the tubing to remain in the
beaker for 30 minutes to allow the glucose to diffuse out
into the medium.
12. Remove the dialysis bag from the beaker. Record the final color
of the solutions in the bag and the beaker in Table 1.
13. Determine the glucose content in the beaker and in the dialysis
bag using glucose indicator strips. To test the solution in the bag,
make a small cut in the bag with a pair of scissors, and insert the
indicator strip through this hole. Record the presence or absence
of glucose in Table 1.
DID YOU KNOW?
Several sea-going birds, such as
penguins and sea gulls, can
drink salt water. They have special glands in their heads that
remove and excrete salt, making
it useful for consumption.
NOTE
Be sure to wash your hands thoroughly before leaving

the laboratory.
B. Osmosis
1. Obtain six plastic cups and label them as follows: water, 0.2 M,
0.4 M, 0.6 M, 0.8 M, and 1.0 M.
NOTE
The pores in the dialysis tubing are extremely small, and

can be easily clogged by any oil or dirt on your fingers
and hands. Wash your hands before handling the dialysis
tubing, and keep physical contact with the tubing to
a minimum.
2. Obtain six pieces of dialysis tubing from the beaker of water. Tie
a tight knot in one end of the tubing, or use a piece of string to tie
off the end.
3. Open one piece of tubing by rubbing the untied end between
your fingers. Pour 25 ml of distilled water into the tubing and
carefully tie a knot in the open end to form a bag. Be sure to leave
enough space in the bag for expansion. Blot the sealed tubing dry
and place it in the cup labeled water.
NOTE
If at any time the dialysis tubing becomes too dry during

the procedure, simply rewet the tubing by placing in water for several seconds and continue setting up
your experiment.
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4. Repeat this procedure with the remaining five pieces of dialysis

tubing, adding a different sucrose solution to each bag: 0.2 M,
0.4 M, 0.6 M, 0.8 M, and 1.0 M. Be sure to blot each piece of tubing dry and place it in the proper cup before proceeding with the
next concentration.
NOTE
5.
Be sure to tie off each of the bags so that they are all approximately the same length.
Carefully blot the bags dry and weigh each one. Record each
bags initial mass in Table 2 in the Analysis section.
6. Fill the six plastic cups approximately 3/4 full of distilled water.
Immerse one bag in each of the cups. Be sure each bag is in the
properly labeled cup.
7.
DID YOU KNOW?

Osmosis is a reversible thermodynamic process. In other
words, the direction of water
flow through a membrane can
be reversed at any time by
proper control of the external
pressure on the solution.
Wait 30 minutes. Remove the bags from the cups. Blot them dry,
and weigh each one again.
NOTE
To ensure accurate readings, the bags must be as dry

as possible.
8. Record the final mass of each bag in Table 2.

9. Calculate the percent change in mass for each of the dialysis bags
using the following formula:
% Change = (Final Mass Initial Mass)/Initial Mass x 100
Record this data in Table 2.
10. Graph both your individual and class average results on the
graph paper in the Analysis section.
C. Water Potential
NOTE
1.
Your instructor will assign you one or more sucrose solutions of varied concentrations and/or distilled water with
which to perform the experiment. Your instructor may
have prepared the potato cylinders in advance. If this is
the case, begin with Step 2.
With the cork borer, cut four cylinders from a potato for each solution you will be using. Cut each cylinder to a length of 3 cm for
greater accuracy; remove any skin from the cylinders. Place the
cylinders in a covered cup or beaker.

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2. Weigh four cylinders together. Record the initial mass of the cylinders in Table 3 in the Analysis section.
3.
Pour 100 ml of one solution you have been assigned into a beaker
or plastic cup. Take the initial temperatures and record in
Table 3. Insert four potato cylinders and cover the beaker or cup
with plastic wrap.
4. Repeat the procedure for the rest of the solutions you will be using, if any. Let the beaker(s) stand overnight.
5. On the following day, measure the final temperature of the liquid
in the beaker(s). Record the temperature(s) in Table 3.
6. Remove the four cylinders from each beaker. Blot them carefully
with a paper towel and weigh them. Record the final mass of the
four cylinders in Table 3.
DID YOU KNOW?
In 1885, Jacobus Henricus
vant Hoff developed a formula to prove that thermodynamic laws are not only valid
for gases, but also for dilute
solutions. His pressure laws
are considered the most comprehensive and important in
the realm of natural science.
7. Calculate the percent change in mass for the four potato cylinders
using the following formula:
% Change = (Final Mass Initial Mass)/Initial Mass x 100
Record this data in Table 3.
8. Graph both your individual and class average results. Place the
percent change in mass on the Y-axis and sucrose molarity on the
X-axis. Using the graph, determine the molar concentration of the
potato cylinders. This is equivalent to the sucrose molarity in
which the potato core mass is constant.
D. Water Potential Calculation

Osmotic potential can be calculated using the following formula:
= iCRT
i = ionization constant (1 for sucrose)
C = osmotic molar concentration (determined in part C)
R = pressure constant (R = .0831 liter bars/mole Kelvin)
T = temperature (Kelvin)
E. Plant Cell Plasmolysis

1. Cut an onion in half lengthwise, from top to bottom, and remove
the center. Note the layers of the scale leaves in the onion. The
epidermal layer, only one cell thick, between each scale is the
layer that will be used in the experiment.

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2. Make several shallow cuts approximately 1 cm apart on one of

the scale leaves. Make more cuts perpendicular to the first set,
resulting in 1 x 1 cm squares.
3. Using forceps, carefully remove the epidermal layer from one of
the squares. If the layer tears while being removed, take another
layer from one of the other squares.
DID YOU KNOW?

In 1901, Jacobus Henricus vant
Hoff received the first Nobel
Prize in Chemistry for his work
in chemical dynamics and osmotic electrical conductivity.
4. Place the layer in two to three drops of distilled water on a microscope slide. Place a coverslip on top.
5. Examine the cell layer under 100X magnification and note the
characteristics of the cells. Draw several representative cells in the
proper space in the Analysis section.
6. Remove the slide from the microscope and add two to three
drops of 15% NaCl solution to one side of the coverslip.
7. Carefully touch the edge of a paper towel to the other side of the
coverslip in order to draw the NaCl solution across the sample.
8. Allow the slide to sit for two to three minutes in the salt solution
and re-examine the sample under the microscope.
9. Note the appearance of the cells. Draw several representative
cells in the proper space in the Analysis section.
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WARDS
AP Biology Lab 1
Lab Activity
Name:
Group:
Date:
ANALYSIS
Table 1
Diffusion
Color
Time
Dialysis Bag
Glucose Content
Beaker
Dialysis Bag
Beaker
Start
30 minutes
Table 2
Osmosis Investigation
Solution
Dialysis Bag
Initial Mass (g)
Dialysis Bag
Final Mass (g)
Change in
Mass (g)
% Change in
Mass
Water
0.2 M
0.4 M
0.6 M
0.8 M
1.0 M
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Table 3
Potato Cell Water Potential
Solution Temperature C
Solution
Initial Temp
Potato Cylinders
Final Temp Initial Mass (g) Final Mass (g)
Change in
Mass (g)
% Change in
Mass
Water
0.2 M
0.4 M
0.6 M
0.8 M
1.0M
Plant Cell Plasmolysis
Cells in Distilled Water
Cells in 15% NaCl
12

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13

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WARDS
AP Biology Lab 1
Lab Activity
Name:
Group:
Date:
ASSESSMENT
1.
Create a Venn diagram comparing osmosis and diffusion.
2.
Part A of the experiment was a demonstration of diffusion. Give an example of diffusion occurring
in the setup. Do you think osmosis occurred in this part of the experiment? If you answered yes, explain why you believe this to be.
3.
Did the dialysis tubing serve as a selectively permeable membrane? Explain your answer.
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4.
In part B, what caused the mass of the dialysis bags to change? Was there more or less water in the
dialysis bags at the conclusion of the experiment? Explain.
5.
Was the distilled water in the beakers hypertonic or hypotonic in relation to the sucrose solutions
found in the dialysis bags?
6.
Suppose the dialysis bags were placed in beakers containing a 0.6 M sucrose solution as opposed to
distilled water. How do you think your results would change? Sketch a graph below to show how
the mass of each of the bags would be affected.
7.
Study the graph you have plotted for part C of the experiment. What is important about the point
where the best fit line crosses the X-axis? What is the concentration of sucrose in your potato?
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8.
Fill in the blanks in the statement below using the following list:
net gain, low, hypertonic, exit, osmotic, enter, less, hypotonic, high, more, net loss, isotonic
(not all words are used)
If a cell is placed in a ____________ solution, it has ____________ solute in solution than the surrounding fluid, and will therefore experience a ____________of water to its surroundings. This cell
has ____________ water potential since there is a great deal of ____________ pressure causing water
to leave the cell. Conversely, a cell sitting in a ____________ solution has a ____________ water potential, and since it will experience a ____________ of water, there will be little osmotic pressure
causing water to ____________ the cell.
9.
Using the data from part C of this activity and the formula for water potential from part D, calculate
the osmotic potential of the sucrose solution in bars.
10. The water potential () of a solution is equal to the osmotic potential () plus the pressure potential
(p). Since there is no differential pressure acting on the solution, the pressure potential is equal to
zero, making the water potential equal to the osmotic potential. If the equilibrium point between the
solutions and the potato cylinders indicates the point where the two water potentials are equal,
what is the water potential of the potato cells?
11. Would the water potential of the potato cells change if the cylinders were allowed to dry out? In
what way?
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12. What are the effects on cells when they are placed in a hypotonic solution, a hypertonic solution,
and an isotonic solution?
13. Why cant humans drink salt water for hydration?
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Diffusion & Osmosis Lab

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TM

DID YOU KNOW?

If a bottle of perfume is opened at one end of the room, in a fairly

2002 WARDS Natural Science Establishment, Inc.

Diffusion also occurs in water separated by a selectively permeable

2002 WARDS Natural Science Establishment, Inc.

DID YOU KNOW?

If a cell is hypertonic to its surroundings, water rushes into the cell,

2002 WARDS Natural Science Establishment, Inc.

DID YOU KNOW?

2002 WARDS Natural Science Establishment, Inc.

Demonstrate diffusion across a semi-permeable membrane

DID YOU KNOW?

Glucose indicator strips

2002 WARDS Natural Science Establishment, Inc.

The pores in the dialysis tubing are extremely small, and

2. Obtain a piece of dialysis tubing that has been soaking in water.

The IKI solution is an irritant; it affects skin and eyes,

8. Note the color of the solution. Record the color in Table 1.

2002 WARDS Natural Science Establishment, Inc.

9. Determine if glucose is present in the beaker by dipping a glucose

Though a color change may become apparent after a few

Be sure to wash your hands thoroughly before leaving

The pores in the dialysis tubing are extremely small, and

If at any time the dialysis tubing becomes too dry during

2002 WARDS Natural Science Establishment, Inc.

4. Repeat this procedure with the remaining five pieces of dialysis

DID YOU KNOW?

To ensure accurate readings, the bags must be as dry

8. Record the final mass of each bag in Table 2.

2002 WARDS Natural Science Establishment, Inc.

D. Water Potential Calculation

E. Plant Cell Plasmolysis

2002 WARDS Natural Science Establishment, Inc.

2. Make several shallow cuts approximately 1 cm apart on one of

DID YOU KNOW?

2002 WARDS Natural Science Establishment, Inc.

2002 WARDS Natural Science Establishment, Inc.

Final Temp Initial Mass (g) Final Mass (g)

Plant Cell Plasmolysis

Cells in Distilled Water

Cells in 15% NaCl

2002 WARDS Natural Science Establishment, Inc.

2002 WARDS Natural Science Establishment, Inc.

Create a Venn diagram comparing osmosis and diffusion.

2002 WARDS Natural Science Establishment, Inc.

2002 WARDS Natural Science Establishment, Inc.

2002 WARDS Natural Science Establishment, Inc.

13. Why cant humans drink salt water for hydration?

2002 WARDS Natural Science Establishment, Inc.

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