Page 1 – Detecting Diffusion Detecting Diffusion Group Leader: ____________________ Materials Technician: _____________________ Recorder: ________________________ Materials Technician: ______________________ Problem: How can you determine whether solutes are diffusing across a membrane? Background A cell membrane is a selectively permeable barrier. Some particles pass through the cell membrane while other particles are held back. Solutes that can move across the membrane generally do so by diffusion. When solutes diffuse they move randomly from an area of higher concentration to an area of lower concentration. When there is no longer a difference in the concentration of solutes in neighboring areas dynamic equilibrium is reached. Solutes continue to move randomly but there is no longer a net movement from high to low concentration. In a cell, diffusion of certain solutes can be facilitated by protein channels that act like tunnels through the cell membrane. Ions and small molecules often move back and forth between the cytoplasm and extracellular matrix using these channels. Conversely, water can move back and forth across the cell’s membrane. When water diffuses, it is called osmosis. This is the movement of water from an area of low solute concentration to an area of high solute concentration. Figure 1 - Diffusion of particles across a selectively permeable membrane In this lab, you will use dialysis tubing to represent the cell membrane. Dialysis tubing is a synthetic membrane used to filter wastes from the blood of a person who has experienced kidney failure. The tubing has small openings, or pores, that allow the passage of relatively small molecules such as water and most ions. Large molecules with a high molecular mass are restricted from passage. Under most conditions, the cutoff is between 12,000 and 14,000 daltons. Other factors such as molecule shape, pH of the solution, and tension on the membrane may affect its permeability. Safety Considerations Iodine solution can irritate the eyes and skin and can stain clothing. Wear gloves, safety goggles, and a laboratory apron while handling any solution that contains iodine. Rinse off any solution that spills on your skin or clothing. Do not direct the points of the scissors toward yourself or others. Use the scissors only as instructed. Page 2 –Detecting Diffusion Pre-Lab Questions Read the entire laboratory, then answer these questions on a separate sheet of paper to be submitted prior to beginning the laboratory. 1. Research the molecular sizes of glucose and starch molecules. Which is larger? Predict which, if either, of these molecules would be able to travel through the pores in dialysis tubing? 2. Recall what occurs when starch solution and iodine solution are added together. How will you know if these two substances come together in this laboratory? 3. Using your answer from #2 above, how will you know whether starch has diffused across the membrane? How will you know whether iodine has diffused across the membrane? 4. How will you be able to tell whether glucose has diffused across the membrane? Materials dialysis tubing Scissors metric ruler 250-mL beakers tubing clamps, string, or twist ties 10-mL graduated cylinder 15% glucose//1% starch solution iodine solution forceps glucose test strips Procedure 1. Obtain and wear goggles, a lab apron, and if you are sensitive to iodine, then you should wear nitrile or latex gloves. 2. Cut a 15-cm length of dialysis tubing. Soak the tubing for one minute in a 250-mL beaker filled with 50 mL of water. 3. Remove the tubing from the water. Fold up 1 cm of the tubing at one end. Use a tubing clamp, twist tie, or piece of string to tightly seal the folded end. 4. Roll the unsealed end of the tubing between your fingers until it opens. Pour 3 mL of 15% glucose/1% starch solution into the tubing. 5. Fold down 1 cm of the tubing at the open end Use a second tubing clamp, twist tie, or piece of string to tightly seal this end. 6. Use tap water to gently but thoroughly rinse the outside of the tubing. Be sure to rinse the clamps or twist ties as well. Do not squeeze the dialysis tubing. Page 3 –Detecting Diffusion 7. Place the tubing in the 250-mL beaker. Fill the beaker with enough water to completely submerge the tubing. 8. Add 4 drops of iodine solution to the water in the beaker. Dialysis tubing with 15% glucose and 1% starch solution 9. Record your initial observations. Use a glucose test strip to measure the glucose concentration in the beaker. Record your results in the data table. 10. Wait 10 minutes, and then record your final observations. 11. Use a forceps to remove and dispose of the tubing as instructed by your teacher. 12. Use your data to answer the Analysis Questions. Data & Results Inside Tubing Color Initial Final Other observations: Is starch Is iodine Is glucose Color present? present? present? Outside Tubing (Beaker) Is starch present? Is iodine present? Is glucose present? Page 4 –Detecting Diffusion Analysis After completing the laboratory answer these questions on a separate sheet of paper. 1. With respect to diffusion, infer what happened to the iodine, starch, and glucose between your initial observations at the start of the experiment and your final observations at the end. 2. Were the predictions that you made for Pre-Lab question #1 accurate? Use what you know about the structure of starch and glucose molecules to explain your results. 3. What substance other than iodine, starch, or glucose moved across the membrane? In which direction did this substance move, and why? 4. Red blood cells are placed in water that has been distilled so that there are no solutes in the water. Are the red blood cells likely to swell up or shrink? Why? 5. The function of the kidney organ in humans is to remove wastes and excess fluid from the blood. If a person experiences kidney failure, they may require dialysis treatment to perform these functions. Research kidney dialysis treatment and explain how dialysis tubing would be used for this treatment. 6. A student performing this lab observed the solution outside of the tubing turning black. What might have happened? 7. Design an experiment to determine if the concentration of glucose inside the dialysis tubing has an effect on its diffusion rate. References: "Diffusion." Unit 4 - Correia Life Science. Web. 11 Oct 2010. <http://www.bio.miami.edu/~cmallery/150/memb/c8.7x11.diffusion.jpg> How Does Dialysis Work? Northwest Kidney Centers, 2014. Web. 13 Dec. 2014. <http://www.nwkidney.org/dialysis/startingOut/basic/howDialysisWork.html>. Kaplan Test Prep. Dialysis tubing in beaker. Learningpod. Learningpod, Inc., 18 Feb. 2013. Web. 13 Dec. 2014. <https://www.learningpod.com/question/ap-biology-question-below-refer-to-the-figure-below-in-which-a-dialysistubing-bag-is-filled/e2b7ced6-5c3f-46b0-961c-39bca96dc6b1>. Miller, Kenneth R., and Joseph S. Levine. Biology-Laboratory Manual A. Teacher ed. Boston: Pearson Prentice Hall, 2010. Print.