Diffusion Through a Membrane State Lab

Part 1 Build a model cell and demonstrate diffusion across

a selectively permeable membrane

Lugols solution (essentially iodine)

*Binds to helical structure of starch (iodines

are the faded purple spheres above), turning
the substance or solution a blue/black, but
only binds because structure is helical.
Therefore, will it bind
glucose?
explain
No,to
it isnt
helical and
therefore
the solution will not turn blue-

Diffusion Through a Membrane State Lab

Part 1 Build a model cell and demonstrate diffusion across

a selectively permeable membrane

Lugols solution (essentially iodine)

What is lugols iodine solution?

Conclusion: Lugols solution is a starch

indicator

Diffusion Through a Membrane State Lab

Part 1 Build a model cell and demonstrate diffusion across

a selectively permeable membrane
1. Making the model cell
Dialysis tubing (see below)
A synthetic membrane that has microscopic pores (holes) that only let
molecules of a certain size or smaller to pass. The one we are using only allows
monomers or smaller molecules to pass.

Diffusion Through a Membrane State Lab

Part 1 Build a model cell and demonstrate diffusion across

a selectively permeable membrane
1. Making the model cell
Dialysis tubing
A synthetic membrane that has microscopic pores (holes) that only let
molecules of a certain size or smaller to pass. The one we are using only allows
monomers or smaller molecules to pass.

If you soak it in water, you

will be able to open up the
tubing to put solution
inside. Before you can do
this, however, you must

Diffusion Through a Membrane State Lab

Part 1 Build a model cell and demonstrate diffusion across

a selectively permeable membrane
1. Making the model cell
Dialysis tubing
A synthetic membrane that has microscopic pores (holes) that only let
molecules of a certain size or smaller to pass. The one we are using only allows
monomers or smaller molecules to pass.

Diffusion Through a Membrane State Lab

Part 1 Build a model cell and demonstrate diffusion across

a selectively permeable membrane
1. Making the model cell
Dialysis tubing
A synthetic membrane that has microscopic pores (holes) that only let
molecules of a certain size or smaller to pass. The one we are using only allows
monomers or smaller molecules to pass.
Glucose, starch, water

Then put
glucose and
starch
solution in
bag

Artificial Cell

Diffusion Through a Membrane State Lab

Part 1 Build a model cell and demonstrate diffusion across

a selectively permeable membrane
2. Place the model cell into a solution containing lugols
(iodine).
Predict what should happen:
1. The glucose should diffuse out down its

I-

I-

Glucose, starch, water

I-

Artificial Cell

concentration gradient and the iodine should

diffuse in down its concentration gradient since
both can pass through the membrane pores. The
starch should stay in too large to pass through
pores. Water should diffuse in and out direction
will depend on how much solute you dissolved in
2. How
will you observe that the iodine
the
water.

I-

went
Thein?
iodine should bind to the helical starch structure

causing the solution inside the dialysis tubing to turn

blue-black.

3. How will you observe that the starch did

not diffuse
The outside
out?
should remain amber (the color of
I-

lugols) since there is no starch present to react with

it.

4. How do we observe that glucose diffused

out?

Diffusion Through a Membrane State Lab

Part 1 Build a model cell and demonstrate diffusion across

a selectively permeable membrane

Use Benedicts solution

- Benedicts is a molecule that reflects blue light that will react with
monosaccharides, lactose and maltose and convert into a molecule
that reflects yellow/orange light. The reaction is endergonic. How we
canYou
we must
get it heat
to happen?
the Benedicts in the presence of the sugars. Input
ENERGY!

1. Add a few
drops of
benedicts to
the solution

2. MUST HEAT!!

3. Turns orange
if
monosaccharid
es, lactose or
maltose is
present

Diffusion Through a Membrane State Lab

Part 1 Build a model cell and demonstrate diffusion across

a selectively permeable membrane
2. Place the model cell into a solution containing lugols
(iodine).
How do we observe that glucose diffused out?

I-

II-

Glucose, starch, water

I-

Artificial Cell

Take a sample of the outer solution into a

clean test tube, add a few drops of benedicts,
and heat it up. If it turns orange then it
diffused out.

I-

I-

Time to do it!
I-

Diffusion Through a Membrane State Lab

Part 1 Build a model cell and demonstrate diffusion across

a selectively permeable membrane

II-

Glucose, starch, water

Q. You have a solution containing 1mM catalase. The pH of the solution is around
6.8 and the salt (NaCl) concentration is around 400 mM (milli Molar). However, you
have previously determined that this protein functions best at a pH of 5.7 and a salt
concentration of 250 mM. Explain how you can get your protein into the desired
conditions without changing the concentration of the enzyme much.
Take a sample of the outer solution into a
clean test tube, add a few drops of benedicts,
IIand heat it up. If it turns orange then it
diffused out.
I

Artificial Cell

I-

Time to do it!
I-

Diffusion Through a Membrane State Lab

Part 1 Build a model cell and demonstrate diffusion across

a selectively permeable membrane
Q. You have a solution containing 1mM catalase. The pH of the solution is around
6.8 and the salt (NaCl) concentration is around 400 mM (milli Molar). However, you
have previously determined that this protein functions best at a pH of 5.7 and a salt
concentration of 250 mM. Explain how you can get your protein into the desired
conditions without changing the concentration of the enzyme much.

Diffusion Through a Membrane State Lab

Part 1 Build a model cell and demonstrate diffusion across

a selectively permeable membrane
Q. You have a solution containing 1mM catalase. The pH of the solution is around
6.8 and the salt (NaCl) concentration is around 400 mM (milliMolar). However, you
have previously determined that this protein functions best at a pH of 5.7 and a salt
concentration of 250 mM. Explain how you can get your protein into the desired
conditions without changing the concentration of the enzyme much.
Catalase solution
with pH 6.8 and
400mM NaCl
Aqueous solution with
pH 5.7 and 250 mM
NaCl

A. Use a dialysis tube. Put your protein

solution into the dialysis membrane and tie it
off. Place the membrane into the desired
solution. Salt will diffuse out and H+ will
diffuse in. Protein will stay inside as it is too
large to fit through pores

This is called Buffer

Diffusion Through a Membrane State Lab

Part 2 Demonstrate diffusion of water across a real
membrane (osmosis)
You want to observe the effect of
salt on osmosis across the plasma
membrane of red onion cells under
the light microscope. How will you
do this?
You look at the cells in water first.
control group
This is your______________.
All of the cells should beturgid
_____________.
What will you do next?

Red onion cells in water (turgid) Add salt to the solution that the
cells are in. What would you
expect to happen and why?

Diffusion Through a Membrane State Lab

Part 2 Demonstrate diffusion of water across a real
membrane (osmosis)

Add salt
(NaCl)

Red onion cells in water (turgid)

plasmolysis

Osmosis (diffusion of water) should occur from inside the cell [high] to outside [low] since the
salt CANNOT cross the membrane and will cause the water concentration outside the cell to
now lower than insideyou should observe plasmolysis. How do we make these cells turgid

Diffusion Through a Membrane State Lab

Part 2 Demonstrate diffusion of water across a real
membrane (osmosis)

Remove
salt (NaCl)

Red onion cells in water (turgid)

plasmolysis

Put them back into tap water (get rid of the salt).

Diffusion Through a Membrane State Lab

Part 2 Demonstrate diffusion of water across a real
membrane (osmosis)

Remove
salt (NaCl)

Red onion cells in water (turgid)

plasmolysis

Remember: This is a real cell membrane (phospholipid bilayer with integral membrane
protein), which will only allow small hydrophobic molecules to pass (you should know
why). Sodium chloride (Na+, Cl-) is charged and hence hydrophilic.