Diffusion and Osmosis Introduction: A cell is a dynamic, living thing. To continue to live, it must continuously exchange materials with its environment, through its cell membrane. In this lab, you will learn about diffusion, one of the major ways that things get in and out of cells. Osmosis is a special example of diffusion, the diffusion of water. Materials: Methyl Orange powder Potassium permanganate crystals Solution of starch and glucose in distilled water Iodine (starch test) solution petri dish containing agar dialysis tubing string 100ml and 10 ml graduated cylinders Procedures: Part 1: Physical diffusion Diffusion is defined as the movement of molecules from an area of high concentration to an area of low concentration without the expenditure of energy. If particles are tightly packed together, their motion will cause them to collide frequently and spread apart. Two factors determine how much and easily particles will spread apart or diffuse. The first factor you will discover in the following experiment. 1. Get a Petri dish containing agar. Draw a line on the bottom of the dish, using a grease pencil, dividing the dish in half. 2. Next, draw a small circle near the outer edge of each section – see figure 1. 3. Using the flat edge of a toothpick, place a small sample of methyl orange onto the agar in one of the little circles. Place an equal sized sample of p potassium permanganate, using another toothpick, into the other circle.. 4. The molecular weight of methyl orange is 327g, and potassium permanganate is 158g. 5. Set the petri dish aside for about an hour, then measure the distance that each chemical diffused from its original edge with your ruler. Record this in your lab. 6. Measurements: a. Methyl orangedidn’t movepotassium permanganatemoved inward 2.3 cm. b. What relationship exists between rate of diffusion and molecular weight? The molecular weight of methyl orange was heavier than potassium permanganate affecting the rate of diffusion. Methyl orange did not diffuse as much as potassium permanganate. Part 2: Dialysis and Osmosis Dialysis is the diffusion of dissolved substances through a differentially permeable membrane while osmosis is the diffusion of water. In this part of the lab you will learn how these processes operate in a cell by making a cell model and subjecting it to a concentration gradient. Dialysis tubing is a cellophane membrane which has been specifically manufactured to contain microscopic pores, and thus to mimic cell membrane. 1. Obtain a 15cm length of dialysis tubing. Soak it in a beaker of tap water for about 30 seconds to soften it. Tie off one end of it very securely with string. 2. Using a funnel, fill the tubing with the glucose-starch solution to within 2cm of the top. Leavean air space in the cell. 3. Very securely tie off the top end of the tube. Check for any leaks, but avoid touching the surface of the tubing as much as possible. 4. Rinse your “cell” under the tap to remove any spilled solution on the outside. 5. Place your “cell in a 100ml graduated cylinder and just cover it with distilled water. 6. Measure 3ml of iodine solution in a 10ml graduated cylinder (Avoid contacting this solution with your hands or clothes), and pour the iodine into the water surrounding your “cell”. Note the time on the clock as you add the iodine. Wait 20 minutes. At the end of 20 minutes observe the results and record them in the data table 1. While waiting for the 20 minutes, continue with the other parts of this lab. Beginning time is: 7. While waiting, get a test tube and add about starch solution to the depth of 1cm. 8. Add one drop of IKI solution to the starch and gently mix the two together. What do you observe? The solution turned black and then faded back to clear. 9. Empty, clean, and return the test tube. 10. When the 20 minutes is up, record your observations in the data table below. Start After 20 minutes Color inside cell clear dark Color outside cell clear dark 11. Based on what you have seen in the “cell” and the test tube, what substance(s) diffused into the cell? Why do you think so? Water and iodine. I think so because the air bubble disappeared and the iodine has stained the inside of the cell a dark black. 12. What substance(s) moved out of the “cell”? Why do you think so? Glucose moved out of the cell because it diffuses from greater to less concentration. 13. You have not tested for the presence of glucose outside the cell. Why would this information be useful? It would determine if glucose has diffused to the outside of the cell. 14. Has the “cell” swollen, shrunk, or stayed the same size? Explain any change observed in terms of osmosis. It was swollen. Water moved into the cell by osmosis. 15. What can you conclude about the size of the pores in the tubing compared to the size of water, starch, glucose, and iodine particles? The size of the pores is smaller than that of starch. But greater than glucose, iodine, and water. 16. This membrane, like cell membrane, is “selectively permeable”. What does this term mean? This term means being permeable to only certain molecules and not all molecules. Before Before After After