TUTORIAL

CELL
PERMEABILITY
Diffusion , Osmosis, and Biological Membranes
This tutorial should be completed AT HOME BEFORE you come into
the laboratory .

Obiectives
When you have finished this tutorial, you should be able to:
1. State in what respects diffusion and osmosis are similar
phenomena and in what respects they differ .
2. Given two solutions of different concentrations separated by a
differentially permeable membrane,
(a) predict in which direction there will be net movement of H2O,
(b) define the relationship between the solutions as
hyper-osmotic, hypo-osmotic, or iso-osmotic.
3. Given a cell--the contents of which are of known concentration--
suspended in one of the above solutions,
(a) predict in which direction there will be net movement of H2O,
(b) define the relationship of the solution to the cell as
hyper-osmotic, hypo-osmotic, or iso-osmotic :
(c) define the relationship between the solution and the cell as
hypotonic, hypertonic, or isotonic .
4. Explain how the forces of osmosis, facilitated diffusion, and
active transport work .' .
5, Describe how a membrane may be differentially permeable to
various molecules.
6. Describe why surface area affects the rate of solute penetration.
- 'DIFFUSION may be defined as the movement of molecules from a
region of higher concentration to one of lower concentration . This
is brought about by tho random motion of the molecules. Therefore,

o
diffusion is a temperature dependent process.
' .Fill
Fill in the two drawings (c,d) showing what would happen to the
of perfume with the passage of time .

OSMOSIS is merely a special form of diffusion. It is primarily

the diffusion of WATER across a differentially permeable membrane .

A differentially permeable membrane is one which will let

some oleculespas while restrictingthepas geofothermolecules.
 Ir. this and subsequent examples, concentrations are- given in
mo It s . This allows us to make direct comparisons in concentrations .
One mole of any substance contains the same number of molecules as
one axle of any other substance . Working with percent, while it
may seem simpler, does not allow us to make such comparisons.
For example:

a.

"solution" of "solution" of solution of

45% beach balls 45% sand particles 4A N.Cl
_8 x 1012 M -8 x 10 -9 M N8 M

If these three "solutions", all at 45% concentration, were

separated by a differentially permeable membrane, in which
direction would the H2O flow?
Check ans. 1

Since osmosis primarily concerns the movement of water

molecules, we are especially interested in the concentration of
water molecules in the solution. When we use molar concentrations,
we can determine the relative concentration of water molecules as
well as of solute particles.

A. B.
For example, container A has 6 x 1023 particles dissolved in
one liter of solution, while container B has only 3 x 1023 particles

A one molar solution has one mole (6 x 1023 particles) of solute

dissolved per one liter of solution .
dissolved per one liter of solution. The concentration of glucose
particles is greater in A than in B. The concentration of WATER
MOLECULES, however, is greater in B than in A!
If the contents of containers A and B were separated by a
differentially permeable membrane, in which direction would
osmosis occur, i.e ., in which direction would the water move'
Check an, 12

Osmotic Pressure
The greater the difference in concentration between the two
solutions, the greater the tendency of the water to move . This
difference in concentration is caused by the NUMBER of particles and
not by the KIND .
This difference may be measured and is called the OSMOTIC
PRESSURE of the solution . In the following diagram, the water
molecules in the right chamber tend to pass through the membrane
and attempt to equalize the water concentration on both sides.

A.

When the weight of the water in the left column exerts a

pressure just equal to that resulting from the tendency of water to
move in, there is no further net change and water moves back and
forth with equal speed. The pressure of the column of water is
termed the osmotic pressure of the solution . It may be defined as
the pressure which exists in a solution because of the tendency of
water molecules to move into that solution in response to an
osmotic gradient .
The solution on the left half of container A has a greater
osmotic pressure than the solution in the right half of container A
since water molecules tend to move from the right to the left .
 a. b.

1> a . In case a, which has the greater osmotic pressure, the cell
or the surrounding medium?

In which direction would water move?

I>b . In case b, which has the greater osmotic pressure, the cell

or the surrounding medium?

In which direction would water move?
Check ans . 8 If right, go to Frame 7 .

If wrong, go to Frame 6 .

Remember : It is the NUMBER of particles and not the kind

which determines the osmotic properties of the solution . A 0.5 M

solution of a molecule which dissociates (like KC1 or NaCI)

produces twice the number of particles as does a 0 .5 M solution of
a molecule that does not dissociate (like glucose) .

7 Osmotic Relationships

SOLUTIONS may be described in terms of their OSMOTIC

RELATIONSHIPS WITH OTHER SOLUTIONS .
.
have Let us suppose that we

differentially permeable membrane bag filled with a salt solution.

We drop it into three solutions :

I
a. b.
 ® Draw an arrow in each case indicating the direction in which
there would be a net movement of water. The answers are below .
In case (a), the water would tend to move from the outside
solution into the bag. In a case like this, the outside solution is
said to be hypo -osmotic to the contents of the bag, i.e ., the
concentration of water molecules outside the bag is greater than
the concentration of water molecules inside the bag.
In case (b), the water would tend to move from the bag to the
outside solution . Here, the outside solution is said to be
hyper- osmotic to the contents of the bag. (The concentration of
H2O molecules outside the bag is less than the concentration of
H2O molecules inside the bag.)
In case (c), there would be no net movement of water because
the concentration of H20 molecules is the same inside and outside
the bag. This solution is said to be iso-osmotic to the contents
of the bag.
Label the following solutions as hypo-osmotic, hyper-osmotic,
I> or iso-osmotic to this membrane-bound solution .

.75M .003M .04m .09M 1.38M

Sucrose Glucose NaCl Cl Sucrose

r
a. . b. c. d. e.

Check ans. 4
 LIVING SYSTEMS : THREE FACTORS INFLUENCING
TRANSPORT OF MATERIALS ACROSS A MEMBRANE

9 Permeability of the membrane

Membranes of living cells are difforentially permeable . To a
certain extent, the capacity of the membrane to restrict the passage
of certain molecules can be described in terms of structural
features, e.g ., proteir-lipid composition, "pores" in the membrane
with varying charge distribution. These features limit the movement
t
of water and solutes along concentration or electrical gradients.
This is the case in simple osmosis, described above. Even after
accounting for these factors, however, slight chemical differences
in different molecules frequently show up as large differences in
ease of penetration. It is apparent that no single factor
determines the permeability'of any membrane to a given substance.
Which of the following can NOT be explained in terms of
simple osmosis?

moves out
H2O moves in H2O moves IN

"an
protein salt

Q.

Check ans. 7

i
Charged particles will tend to move in a direction to equalize
charge differences just as other particles move to equalize
concentration differences.
 Lets stop for a few seconds. So far we have
discussed ONLY THE MOVEMENT OF WATER THROUGH
A MEMBRANE. The SUBSTANCES we have been
considering are unable to pass through the
membrane . Now we shall also be dealing with
COMPOUNDS or PARTICLES THAN CAN PENETRATE THE
MEMBRANE Diagrams a and c in Frame 8
illustrated examples of PARTICLE AND WATER
MOVEMENT through a membrane .
Let us suspend a cell in a solution of glycerol which is
iso-osmotic.

® If the glycerol could not permeate the cell, would you expect
a net movement of water? (Check ans. 2]
But, the cell is permeable to glycerol and the glycerol begins
to diffuse into the cell under the influence of a concentration
gradient (i .e ., outside the cell, the glycerol concentration is
~ 0.3 M and inside the cell it is zero). Each glycerol molecule that

enters the cell contributes to the total number of solute particles

in the cell . This will have the following effect (s) :
® (Check the correct choice(s) .]
a. Osmotic pressure of the cell increases .
b. Osmotic pressure of the cell decreases
c . Osmotic pressure of the cell remains constant
regardless of the influx of glycerol .
d. Water enters the cell .
e. Water leaves the cell .
f. There is not net movement of water.

Check ans . 2
 Tonicity
A cell may swell or shrink even when placed in an iso-osmotic

solution (i .e ., a solution with ORIGINALLY the SAME NUMBER of

particles and water molecules as found in the cell) . This occurs
when the membrane is permeable to the particles in the suspending
solution . The particles will move along a concentration gradient
into the cells (as illustrated below) .

NaCl cannot get out . Glycerol

goes in ;therefore H 20 goes in .

As the particles (i .e ., glycerol) enter the cell and the internal

concentration of total particles (both glycerol and NaCl) increases,
the water flows in immediately to compensate .

In addition to talking about hypo-osmotic , hyper-osmotic , or

iso-osmotic solutions which deal with the concentration of water

and its movement along a concentration gradient, we need another
term to describe the BEHAVIOR OF THE CELL IN VARIOUS SOLUTIONS . To
do this, we use the term TONICITY, TONICITY describes the behavior

of the cell in these various solutions .

a. A hypertonic solution is one which will cause a cell to

shrink .

What is actually causing the shrinkage?

Check ans.

The rate-limiting step in the process is the rate of glycerol

pen etration This step consequently detemines the rate of water
penetration and thus swelling of the cell . [You will determine
this in the laboratory . See Tutorial BI
 b. A hypotonic solution is one which will cause a cell to_ swell .
What is actually causing the swelling?

Check ans. 6b
c. An isotonic solution is one which will cause no change in
the volume of the cells.
®Why is there no change in volume?

Check ans. 6c

Lets stop again for a few seconds .

In summary, remember that :
-OSMOTIC : refers to the movement of
water molecules based upon the
initial number of particles in
the solution .

-'TONIC : refers to the behavior of the

cell in various solutions.

Label the following solutions hypotonic, hypertonic, or

® isotonic to these hypothetical "cells".

Now, label each solution as hypo-osmotic, hyper-osmotic, or

iso-osmotic.
 Facilitated Diffusion and Active Transport :
Forces Acting upon the Substance

We have examined what occurs when simple osmosis is the driving

force behind the movement of substances into and out of the cell .
Additional more efficient transport systems involving "carrier"

proteins or. cell membranes are known to exist . These carriers pick
up materials from the inner or outer membrane surfaces and carry them

across at a faster rate than would occur through simple osmosis .

FACILITATED DIFFUSION is one such type of movement . It DOES NOT

ALTER THE FINAL EQUILIBRIUM! which could be attained even in the
absence of a "carrier system", but it does SPEED UP the attainment of

the equilibrium . Facilitated diffusion does not depend upon

metabolic energy, but the solute carrier-complexes must have

sufficient [kinetic] energy to be able to penetrate the membrane .

ACTIVE TRANSPORT is another type of movement in which a carrier
may be involved . However, it is distinquished from facilitated

diffusion since the passage of the solute through the membrane occurs
AGAINST APPARENT CONCENTRATION AND ELECTROCHEMICAL GRADIENTS at the
EXPENSE OF METABOLIC ENERGY , such as ATP .

The cellular mechanisms of facilitated diffusion and active

transport are not clearly understood .

Label each of the following as simple osmosis or active transport

b. red blood cell c. red blood cell

H2O ,
H2O
0 .2 M 0 .2 M
glycerol glycerol
0 .4 M 0 .3 M
NaCl NaC1

d. How can you distinguish simple osmosis from facilitated

diffusion!

Active transport is affected by metabolic poisons, whereas

facilitated diffusion is not .
13 1 Surface Area
Another factor to consider when dealing with membrane
permeability is a simple question of surface area . Whatever process
is occurring (osmosis, facilitated diffusion, or active transport),
it will be limited by the area of the membrane which is available to
penetration.'
In which of the following membrane-bound figures of the same
total volume would you expect the most rapid osmosis? Why?

0.0 0000 .
0000
0000
00 0000
6.

Check ans. 10

14 n
Summary of Transport in Living Systens
When youare concerned with brane transport, you must
then consider these three factors :
1. The differential permeability of the membrane determines which
substances can pass through and which are excluded. This may
vary with the physiological condition of the membrane, from
cell to cell, and from organism to organism .
2. the f2M which drives the substance--osmotic (chemical
concentration and electrical gradients;, facilitated
diffusion, or active transport .
3 : the surface area of exposed membrane which is available for
penetration.
Little can be inferred unless all factors are accounted for .
In next week's laboratory, you will examine .-,.ore closely some of the
ways in which membrane permeability phenomena can be studied . Be
sure that you see just how each experimental observatin provides
information . As you do the exercises in the laboratory, try to
think of alternative questions which might be asked or procedures
which might be followed
 ANSWER SHEET Tutorial A

1. a - b - c
2. No ; , a,d (If you answered e, think about this . Glycerol has
entered the cell ; therefore its concentration of particles has
risen. If water leaves the cell, its concentration of particles
will go even higher,)
3. Diffusion

4. a. hyper-osmotic b. hypo-osmotic
C. iso-osmotic (remember--2X as many particles in NaCl)
d. hyper-osmotic e. hyper-osmotic
5. s. simple osmosis b. active transport
c. simple osmosis (Glycerol is leaving the cell in response to
a concentration gradient and water follows. See Frame 10
again if you need help .)
d. An equilibrium state would be reached faster through
facilitated diffusion.
Cell shrinks in a hypertonic solution because water leaves .
Cell swells in a hypotonic solution because water enters .
Cell remains the same in an isotonic solution since the
osmotic pressure of the internal and'external solutions are
identical . No net movement of water.
7. a,c
surrounding medium, H2O will move out of the cell .
b. cell, H2O will move into the cell (If wrong, see Frame 6)
9. a. hypotonic (cell swells) ; iso-osmotic (original concentration
of H2O molecules the same inside as outside) .
b. hypotonic (cell swells) ; hypo-osmotic (original concentration
of H2O molecules greater outside than inside) .
c. hypertonic (cell shrinks) ; iso-osmotic (original
concentration of H20 molecules the same inside as outside) .
.10 . C, more surface area in c than in a or b. .
 ANSWER SHEET .
11 . Osmosis occurs in a,b,c. However, there is no NET movement in c.

Explaination . H20 molecules constantly move in both

directions through a differentially permeable membrane . NET
movement will always be from an area of greater concentration
of water molecules to an area of lesser concentration of
water molecules, i.e ., along a concentration gradient .
12 . from b to a.
FOOD FOR THOUGHT '
You and a friend are broiling steak for di^.;:- . Your
friend salts his(her) steak and pops it into the broiler.
After browning it on both sides, he(she) broils it to medium rare .
You brown your steak on both sides, then salt and broil it to
medium rare .
Can you predict a difference in the juiciness of the two steaks?

[If unsure, check Frame 10 .]

Tutorial A
SUMMARY
INANIMATE SYSTEMS

1. Diffusion is the movement of molecules from a region of higher

concentration to one of lower concentration, brought about by
the random motion of the molecules.
2. Osmosis is primarily the diffusion of water across a
differentially permeable membrane .
3. Osmotic pressure is the pressure in a solution which exists
because of the tendency of water molecules to move into that
solution in response to an osmotic gradient .
4. Solutions may be described in terms of their osmotic '
relationship with other solutions :
Hypo -osmotic is a term to describe a situation where the osmotic
pressure outside of the membrane is less than the osmotic
pressure inside of the membrane, i.e ., there is a greater
concentration of H20 molecules outside than inside .
Hyper osmotic is a tern to describe a situation where the
external osmotic pressure is greater than the internal osmotic
pressure, i.e ., there is a greater concentration of H20
molecules inside than on the outside.
Iso-osmotic is a term to describe a situation where the osmotic
pressure of the internal and external solutions are identical .
 SUMMARY-continued
LIVING SYSTEMS -- THREE FACTORS INFLUENCE TRANSPORT OF MATERIALS
ACROSS A MEMBRANE

1. permeability of the membrane is dependent upon membrane

. structure (pores, lipid composition, charge distribution).
Tonicity is a term used to describe the behavior of a cell in a
given solution . .
a. hypotonic--cell swells
b. hypertonic--cell shrinks
c. isotonic--no change in cell volume
2 . Forces acting on the substance :
a. osmosis--movement of water molecules based upon the initial
number of particles in the solution and in
accordance with concentration gradients.
b. facilitated diffusion--movement in accordance with
concentration gradients but more rapid.
c. active transport--movement against concentration and
electrochemical gradients. Requires metabolic
energy.
3. Surface area--rate of penetration dependent upon surface area of
the membrane through which penetration occurs .