/

Page 2 Diffusion and Osmo}is

,/

Objectives:
When you finish this lab, you should be able to: .I
1. Explain the process of diffusion.
2. Explain the process of osmosis.
3. Predict relative rates of diffusion and osmosis in different temperatures.
4. Predict relative rates of diffusion and osmosis in different concentrations.

You are made up of millions of cells. Every single Figure 1: Diffusion

living cell in your body requires food and water in order to
stay alive. 1bis is why you need food and water in order to S•\"-t>t -;,o\\1&.-A\-

stay alive-you must provide for your cells! However, each
cell is surrounded by a plasma membrane·that controls the
movement of materials into and out of the cell. So, how do
materials get in and out of cells?
There are two main categories of transport mecha-
nisms that cells have to move materials through the plasma
membrane: passive and active transport. Active transport
requires the cell to expend energy in order to move ions and
molecules across the membrane. Passive transp.ort does
not-it's considered a "free" process and will occur without
any energy use by the cell. In this lab, you will investigate
two of the mechanisms of passive transport, diffusion and
osmosis.
Diffusion
Diffusion is the movement of particles-ions, mole-
cules or larger compounds-from areas of high concentration
to areas oflow concentration. The difference in concentration
from one area to another is called a concentration gradient.
This movement is a natural consequence of the crowded con-
ditions present in areas of high concentration. The particles
run into each other quite often when all packed together. The
colliding pieces rebound and travel in opposite directions. As
the particles get farther apart, their concentration decreases as
does the rate of collisions. Eventually, all the pieces are
equally distnbuted, and there is no longer a concentration
gradient. The particles are still moving, but there is no net
change in concentration in any area-for every piece moving
to the left, another piece is moving to the right, and so on.
When all particles are evenly distributed, they are at equilib-
rium.
••'~•
o / 0 o
0 ~
0
•
• ••
0 0 0
0 0
• 0 0
• • 0 c>
••
Everyone has experienced diffusion, whether they
realize it or not. If you have ever seen smoke from a fire
spread out in the atmosphere on a calm day, you have wit-
nessed diffusion. If you've ever dissolved sugar or salt in
water, you've practiced diffusion. Diffusion is a spontaneous
process because it does not require energy to get started; in
fact. energy is required to make it stop before it reaches equi-
librium Here's an example:
You have a cat that's mad at you for leaving it alone while
you're in class. Just before you get home, your cat uses the
couch instead of its litter box. As soon as you come in, you
can smell what's happened! If you don't do something soon,
the whole place will smell because the ammonia left by the
cat will diffuse until it reaches equilibrium. To stop the smell
from spreading, you have to clean up the mess-a process
that requires energy.
Environmental Focus: Diffusion
Since diffusion is spontaneous, it will happen wher-
ever a concentration gradient is created. Depending on the
situation, this can be beneficial or harmful to the environ-
ment. Let 's use some examples of diffusion in the environ-
ment to explore this concept further.
Diffusion and Osmosis
Where does diffusion work in the environment? The
short answer would be: everywhere! While that may seem
unlikely, it is mostly true. Have you ever thought about how
flowers are pollinated? Just how do insects know where to
go? Just as you may enjoy the scent of flowers, some insects
use the chemicals created by flowers as a "scent trail" leading
them straight to the flower. The flower creates the chemicals
which diffuse out into the air-from an area of high concen-
tration in the flower to low in the surrounding areas.
That is one way of many in which diffusion works
for the environment, but it can also work against it. What if
there is a paper factory with a storage tank containing chemi-
cal waste built next to a large pond. The storage tank develops
a leak an~ the toxic chemicals are released into the water. The
leak is small, and no one notices the problem until they see
that the plants surrounding the pond are beginning to die. The
leaky storage tank is found and repaired, but when the pond
water is tested, results show that the chemicals have diffused
evenly through the whole pond. Energy will have to be used
in order to reverse the diffusion of chemicals and clean up the
pond.
Although the diffusion of harmful chemicals through
any ecosystem like the pond is not a benefit to the environ-
ment, diffusion can help to ease the harmful effects of pollu-
tion over large areas. Remember, diffusion works on a con-
centration gradient. Part of what detennines concentration is
the size of the space holding the diffusing particles. A larger
area containing the same number of particles as a smaller area
will have a lower concentration of those particles. For in-
stance, a 100mL beaker containing 100 sugar molecules has a
concentration of 1 sugar molecule/mL, while a 500mL beaker
containing 100 sugar molecules has a concentration of 0.2
sugar molecules/mL.
Question 1:
Why do flowers have a stronger fragrance when you stand close to them than they do when you're farther away?
Question 2:
What effect could increasing the concentration of C02 in the USA have on the rate of diffusion of C0 2 worldwide?
Page 3
By now, you've probably heard of global warming
and its relationship with carbon dioxide (C0 2) levels.
col occurs naturally in the environment, but lately concen-
trations have been increasing. The burning of fossil fuels to
power machines like cars is thought to be one of the main
sources of C0 2 contributing to global warming. Millions of
cars have been driven in the US alone for decades, so why is
there such a problem now? Areas like the arctic appear to be
greatly affected by global warming but don' t have cars pro-
ducing carbon dioxide. How is this problem worldwide?
Global wanning is the increase in the average tem-
perature of the surface of the earth. This doesn't just mean
warmer days in summer, it means all year 'round. Warmer
winters can have negative effects on all kinds of plants and
animals. Warmer winters mean that snow will melt faster
and the growing season will start earlier. Animals not ad-
justed to the earlier arrival of flowers and fruit will find
themselves without food. Forests dependent on snowmelt to
feed streams through the summer will have less water in the
hottest months of the year and increase the danger of frre.
Carbon dioxide has always been a part of the at-
mosphere. Currently, the atmosphere is about 0.03% col-
not a large amount! However, C0 2 is very powerful in small
amounts. Changes in even this small amount of C0 2 is of
concern because it absorbs infrared energy (heat) that radi-
ates from the surface of the earth at night and prevents it
from leaving the atmosphere. This trapping of heat leads to
an increase in average temperature over time.
The increase in carbon dioxide is of worldwide con-
cern partly because of diffusion. Even though only some
countries are creating large amounts of C02 , it diffuses from
the areas of high concentration where it is produced to areas
of low concentration where it is not. Therefore, even areas
where there are no people, like the arctic and Antarctic, are
affected by the increase in C0 2 and global warming!
Page 4 Diffusion and Osmosis

Experimental Investigation: Diffusion

Effect of Temp erature on Rate of Diffusion 8. WaH three minutes, and observe the color through the side
of the beaker again. Record your results and answer the
Temperature is the measure of heat. Heat is pro- questions below.
duced by the kinetic energy (movement) of molecules. All
molecules have kinetic energy at temperatures above absolute
Question 3:
zero, so all materials above absolute zero produce heat. In this
In which beaker did the food coloring diffuse the fastest?
experiment, you will determine bow temperature affects the
The slowest?
rate of diffusion.

l. Obtain three 250mL beakers.

2. Label the beakers "1", ''2", and "3".

3. Fill the beaker labeled "1" three-fourths full of cold water.

4. Fill the beaker labeled "2" three-fourths full of room- Question 4:

temper ature water. Explain the differences in temperature of the three beakers in
terms of the kinetic energy of the water molecules.
5. Fill the beaker labeled "3" three fourths full of warm
water.

6. Place the beakers in order on a piece of white paper. If the

beakers are wet, dry them off before placing on the paper.

7. Add one drop of food coloring to each beaker.

Immediately after adding the food coloring, observe what
happens to the color in each beaker by looking through the
side of the beaker-not the top. Record your initial
observations in Table 1 below.

Table 1: Effect of Temperature on the Rate of Diffusion

Beaker 1 Beaker 2 Beaker 3

Water Temperature

Initial Observations

Observations After 3
Minutes
Diffusion and Osmosis Page 5

Osmosis

Osmosis is the diffusion of water across a Figure 2: Main components of a cellular membrane.
selectively permeable membrane. A selectively permeable
membrane is a barrier which only pennits certain particles-
ions, molecules, and larger compounds-to move across from
one side to the other. The selectively permeable membrane
most commonly examined in biological systems is the plasma
membrane which surrounds cells.
The Cell Membrane
To understand how osmosis works, you need to
know what makes up a plasma membrane. The main compo-
nent of a membrane is phospholipids. A phospholipid is a
specialized kind of fatty acid that only has two long lipid
chains instead of three, with the last lipid chain replaced by a
phosphate group. A phospholipid has special properties in the
p resence of water. The lipid part of the molecule is
hydrophobic (water fearing) while the phosphate part is
hydrophilic (water loving). This means that each phosphol-
ipid molecule has two reactions to water at the same time- \.4 Question 5:
is attracted and \.4 is repelled. Molecules that have this dual Why do phospholipids orient themselves so that the hydro-
reaction to water are amphipathic. phobic part is on the inside of the membrane and the hydro-
philic part is on the outside?
Because of their amphipathic nature, phospholipids
naturally form a double-layered membrane when in contact
w ith water. The hydrophobic lipid parts form the inside of the
membrane and the hydrophilic phosphate parts form the outer
and inner surfaces of the membrane. This arrangement forms
a barrier between the environment outside the membrane and
the environment inside the membrane.
The other components of a plasma membrane are
proteins and regular lipids. There are different types of pro- Question 6
teins found in plasma membranes that perform various func- Why is it important for a plasma membrane to be selectively
tions. Some proteins act as channels to allow certain materials permeable?
through the membrane, some form part of a system that al-
lows cells to communicate to each other, and others act as
structural support. The main type of lipid found in a plasma
membrane is cholesterol, which is used in part to control the
distribution of proteins across the membrane as well as for
structural support.
Page 6 Diffusion and Osmosis

Experimental Investigation : Selective Permeability

Selective Permeability
The chemical properties of the molecules that make
up the plasma membrane are what make it selectively perme-
able. Molecules that can make it across a membrane by a free-
energy process like diffusion or osmosis must be small and
not have a strong charge. Charged particles like ions and most
molecules much larger than water cannot pass freely through
the membrane. In the following experiment, you will test the
selectivity of a membrane.
8. Remove the bag and dry carefully, making sure to remove
all excess water. Weigh the bag to the nearest 0.1 g and
record in Table 2.
9. Finish Table 2 by recording your observations about any
changes in the bag and answer the questions.
Question 7:
Did the weight of the dialysis bag change? If so, by how
much?

1. Fill a 400mL beaker with distilled water.

2. Add enough drops ofLugol's Iodine to the beaker to pro-
duce a light brown color. Question 8:
How would a change in the weight of the dialysis bag be
3. Note: either wear gloves or thoroughly wash your hands evidence of osmosis?
before continuing. Take a pre-soaked piece of dialysis
tubing and create a bag by either tying one end shut with
string or using a dialysis tubing clamp. If using a clamp,
fold the last centimeter of tubing over twice and place the
clamp so that it shuts in the middle of the folds.

4. Use a pipette to transfer enough of the starch solution to fill Question 9:

the dialysis bag 3/4 full. Did starch leave the dialysis bag? How do you know?

5. Carefully remove any air that may be in the rest of the bag
and tie or clamp off the open end.

6. Rinse and dry the dialysis bag, making sure to remove all
excess water and weigh to the nearest O.lg. Record the Question 10:
weight in table 2 below. Explain why starch would or would not leave the dialysis
bag.
7. Place the bag in the beaker filled with the diluted Lugol's
Iodine solution and leave the bag in the beaker for 30
minutes before continuing to the next step. Proceed to the
next part of the lab during this time period, but be sure to
come back to this section when the time is up!

Table 2: Selective Permeability

Weight Color of Bag Color of Water in Beaker
Initial Observations

Final Observations
Page 7 Diffusion and Osmosis

Osmosis and Tonicity Question 11:

Did you know that water is called the ''universal Using the diagrams, label each cell as hypertonic, hypotonic,
or isotonic to its environment. Draw an arrow to show the
solvent''? A solvent is a substance in which another material
direction of water movement.
will dissolve. The material that dissolves is a solute.
Together, the mixture of solvent and solute are a solution.
\Vater is called the "universal solvent" because so many com- [ • -:: ~'-'-'~E '-"•'1"•"
0:: )
mon compounds dissolve in water. •
0 • • 0
Since osmosis is the diffusion of water across a
• • • • 0
membrane, like diffusion it is controlled by concentration
gradients. Unlike diffusion, which takes place in a single en- 0 . 0 a •
• 0
vironment, the selectively permeable membrane of osmosis
• Oo 00
creates two separate areas-the inside of a cell vs. its external
• o· 0 0
surroundings. Hav_ing two distinct areas to compare intro-
•
duces three possible directions for a concentration gradient: 0 • o. • 0
#1
If more water is present inside of a cell than outside,
the cell is hypotonic to its surroundings. When a cell is hypo-
tonic, there is a higher concentration of water inside the cell
than outside the cell. Water will follow the concentration gra-
dient and leave the cell, which will shrink as its volume de- 0 • 0
• 0 0 0
creases. 0
#2 • • • •
•
0 •
•
If more water is present outside of a cell than inside,
• • • ••
the cell is hyper tonic to its surroundings. When a cell is hy- 0 •
• 0 •
pertonic, there is a lower concentration of water inside the
cell than outside the cell. Water will follow the concentration 0 • 0 0 0
0
gradient and enter the cell, which will swell as its volume
increases.
#3
If the same amount of water is present outside the
cell and inside, the cell is isotonic to its surroundings. When a
cell is isotonic, there is an equal concentration of water inside
and outs ide the cell. There is no concentration gradient in
isotonic conditions, and there will be no net movement of wa-
0
• 0 • •0 • 0
ter in or out of the cell. 0 0 • •
0
• 0 •
• 0 •
0

• 0 • 0 • •
Diffusion and Osmosis Page 8

Experimental Investigation: Concentration and the Rate of Osmosis

Effect of Concentration on the Rate of Osmosis Question 12:

Which dialysis bag had the greatest weight change?
1. Fill three 400mL beakers with distilled water.

2. Note: either wear g loves or thoroughly wash your hands

before continuing. Take three pieces of dialysis tubing
Question 13:
and make bags as outlined on page 6.
Which dialysis bag had the smallest weight change?
3. Fill the first bag 3/4 full of the blue 10% sugar solution.

4. Fill the second bag 3/4 full of the green 20% sugar
solution.
Question 14:
How did the concentration of sugar in the bag affect the rate
5. Fill the third bag 3/4 full of the red 40% sugar solution.
of osmosis?
6. Rinse and dry each dialysis bag. Be sure to remove all ex-
cess water! Weigh the bags to the nearest O. lg and record
the weights as "Initial Weight" in Table 3 below.

7. Place the bags in the beakers. After five minutes, remove

the bags and dry them thoroughly. Weigh the bags and
record the data.

8. Repeat step 7 every five minutes for a total of20 minutes.

You may wish to set up the next experiment during this
time.

9. When done, discard the dialysis bags as instructed.

10. Graph your results on the following page. To make the

graph easier to interpret, convert your data from weight
into percent change using the following formula:

Weight at TimeX-Initial Weight

- - -- - - - - - - - - X 100 = Percent Change

Initial Weight

Table 3: Con centration and the Rate of Osmosis

Concentration Initial Weight 5 minutes 10 minutes 15 minutes 20 minutes

10%

20%

-W %
Diffusion and Osmosis Page 9

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Diffusion and Osmosis Page 10

Experimental Investigation: Temperature and the Rate of Osmosis

Effect of Temperature on the Rate of Osmosis Question 15:

1. Fill one 400mL beaker with room Which dialysis bag had the greatest weight change?
temperature water, and one with ice water.

2. Fill one beaker with warm water from the water bath and
leave the beaker in the bath. Be sure to label the beaker!
Question 16:
3. Note: either wear gloves or thoroughly wash your hands Which dialysis bag had the smallest weight change?
before continuing. Take three pieces of dialysis tubing
and make bags as outlined on page 6.

4. Fill each bag 3/4 full of the red 40% sugar solution.
Question 17:
5. Rinse and dry each dialysis bag. Be sure to remove all ex- How did the temperature of the water affect the rate of osmo-
cess water! Weigh the bags to the nearest O.lg and record sis?
the weights as "Initial Weight" in Table 4 below.

6. Place the bags in the beakers. After five minutes, remove

the bags and dry them thoroughly. Weigh the bags and
record the data.

7. Repeat step 6 every five minutes for a total of20 minutes. Question 18:
You may wish to set up the next experiment at this time. Would not leaving the warm beaker in the water bath have
affected your results? Why?
8. When done, discard the dialysis bags as instructed.

9. Graph your results on the following page. To make the

graph easier to interpret, convert your data from weight
into percent change using the following formula:

Weight at TimeX- Initial Weight

- -- - - - -- - - - - X 100 = Percent Change

Initial Weight

Time X is your recorded weight at 5 minutes, 10 minutes, etc.

Table 4: Temperature and the Rate of Osmosis

Temperature Initial Weight 5 minutes 10 minutes 15 minutes 20 minutes

Cold

Room

Warm
.·

Page 11 Diffusion and Osmosis

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Page 12
Environmental Focus: Osmosis
"Water, thou hast no taste, no color, 110 odor; canst not
be defined, art relished while ever mysterious. Not
11ecessary to life, but rather life itself, thou jillest us with
a g ratification that exceeds the delight ofthe senses."
Antoine de Saint-Exupery, From Wind, Sand and Stars, 1939
Water is a crucial component of every living thing.
Since osmosis is a spontaneous process, anything that affects
water concentrations around cells will change their water con-
tent. Alterations in water concentration or solute concentra-
tion change the osmotic pressure inside the cell. Osmotic
pressure depends on the strength of the concentration gradi-
ent; the larger the concentration gradient, the higher the os-
motic pressure.
Have you ever thought about why you get thirsty?
When the cells of your body don't have enough water, this
has the effect of raising their solute concentrations, increasing
their concentration gradient and osmotic pressure. This in-
crease sends signals to your brain that you translate as a sen-
sation of thirst. When you drink water, you don't feel thirsty
anymore because you've increased the water concentration
inside your cells, reducing osmotic pressure.
Animal cells can have problems when solute concen-
trations change. All that separates an animal cell from its sur-
roundings is the plasma membrane, which is permeable to
water . When the solute concentrations in a cell's environment
change, the cell cannot control the movement of water across
its membrane. In order to keep the proper balance of water,
the cell must expend energy to either increase or decrease its
water content. Otherwise, the cell can swell up until it bursts
(lysis) pr shrink (crenation).
M~y animals have specialized ways for dealing
Question 19:
Why is it dangerous to drink too little or too much water? Explain your answer with reference to lysis and crenation.
Question 20:
Once a piece ofland has been "salted", why is it so hard to make it fertile again?
Diffus ion and Osmosis
with changes in osmotic pressure. Single-celled organisms
like Amoeba have contractile vacuoles, specialized organ-
elles that collect excess water from cytoplasm and remove it
from the cell. Multicellular organisms have even more com-
plex systems for water management; these are usually called
urinary systems.
Plant cells are affected by changes in osmotic pres-
sure just like animal cells, but their plasma membranes are
surrounded by a rigid cell wall. Let's use the following exam-
ple to illustrate the effect on plant cells of changing solute
concentrations in the soil:
For thousands of years, humans have used osmosis
as a part of warfare, punishment, and as a way to have ·a fmal
say in territorial disputes. Salting the earth has been a
method of destroying the fertility of cropland and making
areas unlivable for long periods of time by severely altering ·
the solute concentration of soil and groundwater systems.
How does salting the earth turn land that was once full of
green fields into a virtual desert?
Plants get water from the soil through osmosis. The
cells in the roots of plants are usually hypertonic to ground
water, so water will freely enter the root cells. By salting the
earth, solute concentrations in the soil and groundwater are
greatly increased, and water concentrations decrease in re-
sponse. This makes the plant's root cells hypotonic to their
environment. Water won't enter the cells, instead the cells
will lose water. Unlike animal cells the plant cells won't
shrink because of their cell wall, but the plasma membrane
will shrivel, a process called plasmolysis. When this occurs
in enough cells, the plants will lose turgor pressur e and
wilt. If water concentrations do not increase, the plants will
dehydrate and die.
.·.
.