Diffusion and Osmosis

Introduction:

Atoms and molecules are the building blocks of cells. Both have kinetic energy and are constantly in motion. They continually bump into one another and bounce off into new directions. This action results in two important processes, diffusion and osmosis.

Diffusion is the random movement of molecules from an area of higher concentration of those molecules to an area of lower concentration. Cells have selectively permeable membranes that only allow the movement of certain solutes. Diffusion is vital for many of lifeís functions in a cell. It allows oxygen and carbon dioxide exchange in the lungs and between the bodies of intracellular fluid and cells. Diffusion also aids in the transport of nutrients and water in the xylem and phloem of plants. In those plants, it permits for the absorption of water into roots. An example of this process is the diffusion of a smell in a room. Eventually dynamic equilibrium will be reached. This means that the concentration of the molecules carrying the smell will be approximately equal through out the surrounding enclosed area and no net movement of the molecules will occur from one area to another.

Osmosis is special kind of diffusion. It is the diffusion or movement of water through semi-permeable membranes from a region of higher water potential (hypotonic solute) to a region of lower water potential (hypertonic solute). Water potential is the measure of free energy of water in a solution. There are three types of solutions. Isotonic solutions have an equal concentration of solute on both sides of the membrane, and dynamic equilibrium has been reached in the solution. Hypertonic solutions have a higher concentration of solute on one side of the membrane than the other. Hypotonic solutions are the opposite of hypertonic solutions. A solute is what is being dissolved by the solvent (water is the most common solvent) in a solution.

Water will always move from an area of higher water potential to an area of lower water potential. An important factor effecting of diffusion and osmosis is water potential. Water potential measures the tendency of water to leave one place in favor of another place. Water potential is affected by two physical factors. One factor is the addition of solute, which lowers the water potential. The other factor is pressure potential. An increase in pressure raised the water potential. The water potential of pure water at atmospheric pressure is defined as being zero. The Greek letter psi is used to represent water potential. The following formula can be used for calculations:

ψ (Water potential) = ψp (Pressure potential) + ψs (Solute potential)

Movement of water into and out of a cell is influenced by the solute potential on one side of the cell membrane relative to the other side. Plasmolysis is a phenomenon in walled plant cells in which the cytoplasm shrivels and the plasma membrane pulls away from the cell wall when the cell loses water to a hypertonic environment. This leads to a loss of turgor pressure (the force directed against a cell wall after the influx of water and the swelling of a walled cell due to osmosis) and eventual death of the plant. If water moves into the cell, the cell may lyse, or burst (in animal cells, plant cells are equipped to handle large intakes of water). Water movement is directly proportional to the pressure on a system. Pressure potential is usually positive in living cells and negative in dead ones.

Diffusion and osmosis are not the only processes responsible for the movement of ions or molecules in an out of cells. Active transport is process that uses energy from ATP to move substances through the cell membrane. Normally, active transport moves a substance against its concentration gradient, that is to say from a region of low concentration to an area of higher concentration.

Hypothesis:

Osmosis and diffusion will continue until dynamic equilibrium is reached and net movement will no longer occur. Diffusion is effected by the solute size and concen-tration gradient across a selectively permeable membrane. Water potential greatly determines the results in sections of the experiment.

Materials:

Exercise 1A

For this exercise, the following materials are required: a 30 cm of 2.5 cm dialysis tubing, 250 ml beaker, distilled water, funnel, 2 dialysis tubing clamps, 15 ml of 15% glucose/1% starch solution, 4 pieces of glucose tape, 4 ml of Lugolís solution (Iodine Potassium-Iodide or IKI), a timer, paper and pencil.

Exercise 1B

This exercise of the experiment requires six strips of 30 cm dialysis tubing, 250 ml beaker, 12 dialysis tubing clamps, funnel, six cups, distilled water, an electronic balance, timer, paper towels, and about 25 ml of each of these solutions: distilled water, 0.2 M glucose, 0.4 M glucose, 0.6 M glucose, 0.8 M glucose, and 1.0 M glucose. For recording results, paper and pencil are necessary.

Exercise 1C

This part of the experiment requires a large potato, potato corer (about 3 cm long), 250 ml beaker, paper towels, scale, six cups, knife, paper, pencil and about 100 ml of each of these solutions: distilled water, 0.2 M glucose, 0.4 M glucose, 0.6 M glucose, 0.8 glucose, and 1.0 M glucose.

Exercise 1D

This section requires a calculator, paper, pencil, and graphing paper.

Exercise 1E

This section of the experiment requires paper, pencil, paper towels, onionskin, dye, microscope, slide, cover slip, salt water (15%), and tap water.

Methods:

Exercise 1A

First, soak the dialysis tubing in distilled water for 24 hours. Before handling the tubing, wash dirty hands thoroughly to prevent getting oils on the dialysis tubing and changing the results. Remove the tubing and tie off one end using the clamp. To use the clamp, twist the end of the bag several times and then fold it onto itself. Next, open the other end of the tubing by rubbing the end between two fingers. Fill it with the glucose/starch solution using a funnel. Use the glucose tape by dipping it into the solution. Record the color change of the tape and the color of the bag. Tie of the end with the tubing clamp. It is necessary to leave space for expansion but no air. Fill the beaker with distilled water and add the 4-ml of Lugolís solution. Record the color change. Use glucose tap to test for any glucose in the water. Record these results. Set the dialysis tubing in the beaker and let it sit for about 30 minutes. Remove the bag and record the change in water and bag color. Use the last two pieces of glucose tape to measure the glucose in the water and bag. Record results.

Exercise 1B

First, soak the dialysis tubing for about 24 hours. Again be sure to cleanse hands. Tie off one end of each tube with the clamps. Next, fill each tube with a different solution (distilled water, 0.2 M glucose, 0.4 M glucose, 0.6 M glucose, 0.8 glucose, and 1.0 M glucose) with the funnel and tie off the end again leaving empty space, but no air. Weigh each bag separately on the electronic balance and record the masses. Soak the bags in separate cups filled with distilled water for about 30 minutes. Remove the bags and gently blot dry with paper towel. Reweigh, and record the mass.

Exercise 1C

First, slice the potato into to 3-cm discs. Use the potato corer to core out 24 cores. Weigh 4 cores together and record their mass. Fill each cup with one of the following solutions: distilled water, 0.2 M glucose, 0.4 M glucose, 0.6 M glucose, 0.8 glucose, and 1.0 M glucose. In each cup put 4 potato cores, and allow them to sit over night. Take out the cores and blot them dry. Again weigh them on the electronic scale. Record the change in mass. Calculate the information for the table. Compare the results with another group.

Exercise 1D

First, determine the solute potential of the glucose solution, the pressure potential, and the water potential. Graph the information given about the zucchini cores.

Exercise 1E

Prepare a wet mount slide of dyed onion skin. Observe under a light microscope and sketch how the cells appear. Add a few drops of the salt solution using a paper towel to wick the solution under the slip. Observe how the cells are effected and make another sketch.

Results:

Exercise 1A

 

Table 1: Change of Color of Dialysis Tubing and Beaker

Solution Color

Presence of Glucose (Glucose Tape)

Initial

Final

Initial

Final

Dialysis Bag

15% Glucose/1% Starch

Milky White

Midnight Blue

Algae Green

Mahogany

Beaker

Water + IKI

Amber

Rusty Amber

Pear Green

Olive Green

Initial Glucose Tests Final Glucose Tests

 

Which substance(s) are entering the bag and which are leaving the bag? What experimental evidence supports your answer? Iodine Potassium Iodide and water enter the bag. This is proven by the color change (starch test) and the increase in the size of the bag. Glucose left the bag and this is proven by a positive test on the surrounding water.

Explain the results you obtained. Include the concentration differences and membrane pore size in your discussion. The results show that the water, glucose, and IKI molecules were small enough to pass through the selectively permeable membrane. The starch didnít leave the beaker because its molecules were too large to pass through the selectively permeable membraneís pores.

Quantitative data uses numbers to measure observed changes. How could this experiment be modified so that quantitative data could be collected to show that water diffused into the dialysis bag? The bags could be massed before and following their immersion in the solution. The volume of the solution in the beaker could be found before and after the immersion of the bag by using a graduated cylinder.

Based on your observations, rank the following by relative size, beginning with the smallest: glucose molecules, water, IKI, membrane pores, and starch molecules. The smallest substance was water, then the IKI molecules, glucose, the membrane pores, and the largest substance was the starch molecules.

What results would you expect if the experiment started with a glucose and IKI solution inside the bag and only starch and water outside? Why? Based on the size of the molecules, the glucose and IKI would move out of the bag and the water would go in. The large starch molecules would be left in the beaker.

Exercise 1B

Table 2: Dialysis Tubing Mass Change Results: Individual Data

Contents of Dialysis Tubing

Initial Mass (g)

Final Mass (g)

Mass Difference (g)

Percent Change in Mass

a) Distilled Water

26.0

26.2

0.2

.77%

b) 0.2 M

27.0

27.5

0.5

1.85%

c) 0.4 M

25.0

25.6

0.6

2.4%

d) 0.6 M

27.9

31.4

3.5

12.54%

e) 0.8 M

28.3

32.0

3.7

13.07%

f) 1.0 M

28.4<