BIOL-111: Lab 8 – Membrane Structure & Transport

Use your lab handout and other resources provided to complete the following worksheet activity.

Part 1: Label the membrane components.

Use the following words to label the cell membrane components and structures:

(Image modified from Cengage Learning- 2011, 2015, 2019)

Carbohydrate groups Glycolipid Integral Proteins Phospholipid Polar head

Cholesterol Hydrophobic region of protein Peripheral Protein
Cytoskeleton Hydrophilic region of protein Phospholipid hydrophobic tails

A. F.
B. G.
C. H.
D. I.
E. J.

What is the fluid mosaic model for the plasma membrane? Why is the membrane fluid?

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Part 2: Membrane Transport – Diffusion and Osmosis

Complete the following crossword puzzle to get familiarized with the diffusion and osmosis terms.

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Part 3: Diffusion Experiments

Experiment #1 – Brownian Motion
Watch the following video about Brownian motion  Brownian Motion Video

1. What is Brownian motion?

2. When looking at the small particles in the video, do the particles move in random directions?

3. Do particles of different size move at the same or different speeds?

Experiment #2 – Diffusion Rate

You move on to study diffusion of particles with different molecular weights. You take Petri dish filled
with agar, make two holes in the agar and fill one hole with methylene blue and the other one with
potassium permanganate.
Potassium Permanganate (PP) = Molecular Weight = 158
Methylene Blue (MB) = Molecular Weight = 320
Agar gel = gelatin consisting of 98.5% water and freely permeable to PP and MB.

Watch the experiment video  Diffusion Rate Video

Here are your experiment results. Carefully observe your results and answer the questions below.

(Images courtesy of Dr. Rohrer, 2019.)

1. Which substance diffused faster?

2. Was it the substance with the higher or lower molecular weight?

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3. Based on this result, what is the relationship between molecular weight and rate of diffusion?

Experiment #3 – Diffusion of Gases

In the next experiment, you are studying the diffusion of gases. You are going to observe the diffusion
of ammonium hydroxide (NH4OH) and hydrochloric acid (HCl) in a glass tube. The molecular weight for
NH4+ ions is 18 and for Cl- ions is 35.5.

1. Based on this information, what would your hypothesis be?

You place a small amount of each substance at opposite ends of a glass tube. Each gas diffuses towards
the center of the tube. HCl will form H+ and Cl-. When the NH4+ and Cl- meet, they will produce NH4Cl,
which forms a white precipitate. The results of your experiment are shown below.

(Images courtesy of Dr. Rohrer, 2019.)

2. Based on the results shown here, which substance will diffuse faster, particles with lower
molecular weight or particles with high molecular weight?

3. Is this result consistent with your observations of the rate of diffusion in the agar plate?

Part 4: Osmosis Experiments

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Experiment #4 – Diffusion across semi-permeable membrane- Effect of varying concentration.

After learning about the process of osmosis, the diffusion of a solvent, you now move the test the
concept. With this experiment you will see how the relative osmolarity of the solutions on either side
of a membrane affects the rate of osmosis. You fill dialysis bags with either sucrose solutions of
varying concentration or water and then immerse the bags in either water or sucrose solution. At the
end, you will measure the movement of water by measuring changes in the weight of each bag. You
can find more details about the experiment in the lab handout.

Watch the following video about a similar experiment set up.  Diffusion across dialysis bag video.

Below you can see the setup for the lab experiment:

(Images courtesy of Dr. Rohrer, 2019.)

After performing the experiment, you obtain the following results:

  1 2 3 4
Bag 1 = dH2O Bag 2 = 15% sucrose Bag 3 = 30% sucrose Bag 5 = dH2O
Time Beaker = dH20 Beaker = dH20 Beaker = dH20 Beaker = 30% sucrose
(min.) Mass ∆ Mass Mass ∆ Mass Mass ∆ Mass Mass ∆ Mass
(g) (g) (g) (g) (g) (g) (g) (g)
0 10.1   10.0   10.0   10.0  
15 10.05 0.05 10.3 0.3 10.6 0.6 9.3 -0.7
30 10.0 0.0 10.6 0.6 11.2 1.2 8.7 -1.3
45 10.0 0.0 10.9 0.9 11.8 1.8 8.2 -1.8
60 10.0 0.0 11.2 1.2 12.2 2.2 7.9 -2.1
(Table modified from Rohrer, William and Zanes, Everett. Bio111-Lab 8-Diffusion Lab Handout. Union County College. Cranford, NJ.)

Using the change in mass calculated in the table above, answer the following questions:
1. How is the water movement in each of the bags? (Is water moving out or inside of the bag).
Bag 1:

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Bag 2:
Bag 3:
Bag 4:

2. Determine the tonicity of the solution (isotonic, hypotonic, hypertonic):

Bag 1:
Bag 2:
Bag 3:
Bag 4:

3. What can you conclude from this experiment?

Osmometer Demonstration.

For the following experiment, you

will measure the movement of
water across a semi-permeable
membrane using two osmometers.

The osmometer on the left contains

15% sucrose solution and the one
on the right contains 30% sucrose
solution. Water will move by
osmosis from the beaker into the
bag and then move up the capillary
tube. The capillary tube has marks
that allow you to measure the
volume of fluid that enters.

1. In which osmometer, do you

predict a faster rise in the water
level?
(Images courtesy of Dr. Rohrer, 2019.)

Part 4: Hemolysis Test.

Human red blood cells are sensitive to osmotic changes. For example, when they are placed in a
hypotonic solution they undergo hemolysis. The hemolysis test allows us to see if the plasma

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membrane of a red blood cell is intact or has been ruptured by osmosis. If a suspension of cells are
intact (isotonic or hypertonic solution), it will look cloudy. A suspension of hemolyzed cells will have a
clear appearance and will allow light to pass through.

Hemolysis Test:
After suspending red blood cells in
solution, hold up the test tube
containing the blood suspension in
front of a page of printed text.
 If the text IS NOT legible
(solution cloudy) = the blood
cells are intact (solution is
isotonic or hypertonic).
 If the text IS legible (solution
clear) = hemolysis has
occurred (solution is
hypotonic).
(Images courtesy of Dr. Rohrer, 2019.)
Watch this video to see what happens to the red blood cells during hemolysis.  Hemolysis Video.

Experiment #5 – Alcohol Diffusion

For the next experiment, you will use the hemolysis test to determine whether three different alcohols,
which differ in their lipid solubility, differ in their ability to cross the plasma membrane.

The partition coefficient compares the solubility of a substance in oil to its solubility in water. The
partition coefficient will be larger for substances that are more lipid soluble. This means that molecules
considered to be relatively non-polar and more hydrophobic will have larger partition coefficients than
relatively polar and more hydrophylic molecules.

1. Which has the most carbon and hydrogen atoms relative to the number of oxygen atoms?

2. Based on this, which is most non-polar and has the largest partition coefficient?

3. Based on the partition coefficient and your knowledge of the structure of each alcohol, which
alcohol would you predict has the greatest effect on the membrane?

Tube Alcohol Formula Partition Time until hemolysis

Coefficient (Seconds)
1 Methyl alcohol (methanol) CH3OH 0.010 17

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2 Ethyl alcohol (ethanol) C2H5OH 0.036 9

3 Propyl alcohol (propanol) C3H7OH 0.156 4

(Table modified from Rohrer, William and Zanes, Everett. Bio111-Lab 8-Diffusion Lab Handout. Union County College. Cranford, NJ.)

The three videos presented below show the hemolysis test for the three alcohols listed in the table
above. You will see that the solution becomes cloudy when blood is added, which makes it difficult to
read the text in the back of the tube. After few seconds, the solution becomes clear, making it easier to
see the text in the back. It is not easy to determine exactly when the solution becomes clear. The
table shows data collected from previous semesters. Click on each image to watch the video.

Methyl Alcohol Ethyl Alcohol Propyl Alcohol

(Videos
courtesy of Dr. Rohrer, 2019.)

4. Which alcohol diffuses across the membrane fastest?

5. your results support the expected relationship between lipid solubility and membrane
permeability?

6. Based upon your results, will Hexanol (C6H13OH) cross the membrane faster, or slower, than
propanol (C3H7OH)? Briefly explain your answer (Why yes or no?)

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Experiment #6 – Covalent and Ionic Compounds and Their Impact in Osmotic Pressure.
In this last experiment you are going to compare the response of cells to glucose and NaCl solutions
which have the varying molar concentration. Ionic and covalent compounds differ in the number of
solute particles that they form in a solution, and thus, this affects their impact on the osmotic pressure
of the solution. For example, covalent compounds don’t ionize with water, while ionic compounds can
ionize in solution producing 2 or more particles. Therefore, each type of compound with have a
different effect in the osmotic pressure of a solution.

For this experiment you are going to use the Hemolysis Test to determine the concentration of glucose
(covalent compound) and of NaCl (ionic compound) that results in an isotonic solution, respectively.
Following the protocol in your lab handout, you obtain the following results:

(Images courtesy of Dr. Rohrer, 2019.)

Based on these results, complete the following table by addressing the questions below:
1. Is it cloudy or clear? (based on the description provided)
- Type in the table: Cloudy or Clear
2. Are the cells intact or has there been hemolysis? (your conclusion based on the description and
your knowledge about the Hemolysis Test)
- Type in the table: Intact cells (if no hemolysis) or Lysed cells (if hemolysis)
3. Is the solution hypertonic, isotonic, or hypotonic? (your conclusion based on your
understanding of osmolarity and its effect on cells)
- Type in the table hypertonic, isotonic or hypotonic.
- Refer to your lab handout for more details if needed.

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Tube Number and Molarity of the solution

Decreasing solute concentration

Tube 1 Tube 2 Tube 3 Tube 4 Tube 5 Tube 6 Tube 7

Solution 1 1 1 1 1 1 1
/6 M /8 M /10 M /12 M /14 M /16 M /18 M

Glucose

NaCl

(Table modified from Rohrer, William and Zanes, Everett. Bio111-Lab 8-Diffusion Lab Handout. Union County College. Cranford, NJ.)

Based on your table, respond the following questions:

1. At what concentration does Glucose becomes isotonic in solution?

2. At what concentration does NaCl becomes isotonic in solution?

3. Which isotonic solution (Glucose or NaCl) has the higher solute concentration? Briefly explain
your answer.

4. If you had to prepare an isotonic solution of KCl, what molar concentration should it have based
upon your results?

5. If you had to prepare an isotonic solution of maltose, what molar concentration should it have
based upon your results?

References:
Biology Corner. https://www.biologycorner.com/worksheets/diffusion_osmosis_crossword.html
Dr. Rohrer, 2019 materials.

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Cengage Learning-Chapter 5. 2011, 2015, 2019.

Rohrer, William and Zanes, Everett. Bio111-Lab 8-Diffusion Lab Handout. Union County College. Cranford, NJ.)

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