Diffusion across Biological Membranes: A simulation

Introduction Diffusion is the process by which collisions between molecules cause to spread apart. This movement is described as movement from an area of greater concentration to an area of lower concentration. Hence, diffusion continues until the molecules are equally distributed. This is to ensure that molecules have reached a state of equilibrium. Diffusion occurs spontaneously, no energy is involved. In cells, Diffusion occurs through cell membrane which is selectively permeable. This means that some molecules can pass through the membrane while other molecules cannot. Molecules that move across the membrane through diffusion are either hydrophobic or small and uncharged. For example, H2O, O2 and CO2. On the other hand, those molecules which did not make through the membrane such as large polar molecules must be transported through protein mediated mechanisms to cross the membrane. Hence, this experiment is focused on diffusion and required to create a own model cell by pouring distilled H2O, starch, iodine and glucose in tied dialysis tubing to test for substance which diffuse through the membrane and which one could not due to its size.

Materials and Methods Materials: Distilled H2O Starch solution Iodine solution Glucose solution Test tube (3-4/group) Test tube rack Marking tape Benedicts test solution Hot water bath Dialysis tubing: 1 inch flat width, pre-soaked Dialysis tubing: 3 inch flat width, pre-soaked Breaker 3ml pipette

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Methods: Tract the substance Step 1: Labeled 4 test tubes as Distilled H2O, Starch, Glucose and Iodine. Step 2: Add 2ml of distilled H2O, Starch, Glucose and Iodine in the respective test tubes and add 2ml of Iodine indicator into all 4 test tube. Step 3: Swirl the test tubes lightly to ensure the Iodine indicator and 4 solutions mixed well. Observed the colour change closely as shown in table 1.0 Step 4: After Iodine test, rinse the test tube with distilled H2O to ensure all traces of the solution is being washed away and neutralize any charges present. Step 5: Repeat step 2 with 2ml of Benedicts solution and swirl the test tubes lightly and observe the colour change closely as shown in table 1.0

Cell simulation Step 6: Take 1 inch dialysis tubing and fasten one end of the tube tightly. Step 7: Pour full of iodine solution and full of glucose solution and fasten the other end tightly as well. Step 8: After which, run 1 inch dialysis tubing under distilled water. Step 9: Take 3 inch dialysis tubing an repeat step 6. Step 10: Pour starch solution almost full and submerge 1 inch dialysis tube into 3 inch dialysis tube. Fasten the other end tightly. Again rinse 3 inch dialysis tube with distilled H2O. Step 11: Fill the beaker with H2O and submerge the dialysis tubing in and set aside for 1 hour. Step 12: After 1 hour, observe the colour change.

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Results Chemical Test Result

Table 1.0, Chemical test results.

Table 1.0 shows the result of distilled H2O, Starch, Glucose and Iodine performed using Benedict solution and Iodine test. The left column is the 4 different substances which are used to represent cellular components. The right column is the 2 indicators used; Iodine indicator and Benedicts. Referring to the table, when both indicators were added to distilled H2O no change occurred. When starch was added to iodine indicator dark blue precipitate was formed. Similarly when starch was added to Benedicts and left it in water bath 3minutes, blue precipitate was formed. When glucose was added to iodine indicator, no change occurred. On the other hand, when glucose was added to Benedicts and left it in water bath 3minutes, brick red precipitate was formed. Lastly, when iodine was added to benedicts with 3 minutes of water bath there were no change occurred.

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Cell Simulation

Figure 1 Design of own cell simulation

Drew the design of our cell simulation as seen on figure 1, and build the cell using 1 inch and 3 inch dialysis tube consist of iodine and glucose and starch respectively. The dialysis tube was submerged in a beaker filled with water. Both dialysis tubes were rinsed by distilled H2O before placing them inside and left the cell model aside for one hour. After one hour, the 3 inch dialysis tube which carried starch turned dark blue precipitant.

SSSSSSS SSSSSSS gigigigig gigigigig H2O s s s s

Si iS i iSi SiS

gigigi H2O s i s s i
Final Stage

Figure 2, Diffusion of molecules before and after.

From figure 2, we can observe diffusion taking place.S represent starch, i represents iodine and g represent glucose. As initially, 1 inch dialysis tube consists of iodine and glucose and 3 inch dialysis tube consists of starch. However, after 1

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hour, the iodine molecules got diffused into 3 inch dialysis tube and also the water in the beaker turned slightly cloudy in the final stage.

Figure 3, the current location of each substance after1 hour

As changes were observed due to diffusion to confirm, chemical test was conducted with solution outside the cell, inside of cell and within organelle. Firstly, extracted solution outside of cell and did Iodine test and Benedicts test. Iodine test was conducted to check for the presence of starch by adding 2 ml of iodine indicator into 2 ml of extracted solution outside of cell. However, there were no visible changes observed. Subsequently added 2ml of benedicts solution to 2ml of extracted solution outside of cell and placed them in water bath for 3minutes. The result obtain was cloudy Therefore assume there was a presence of glucose while absence of starch and iodine as shown in figure 3. Secondly, extracted solution inside of cell and did the same tests as earlier to detach the presence of starch, iodine and glucose. 2ml of the solution was added to 2ml of iodine indicator to test for presence of starch. Dark blue colour change was observed through the test. Subsequently, 2ml of benedicts solution was added with 2ml of solution extracted inside of cell to test for presence of glucose and there was no visible changes observed. Hence, there were starch and iodine present inside of cell as shown in figure 3 Lastly, extracted solution within organelle by opening up the tube and added 2 ml of starch solution to 2 ml of extracted solution. Blue colour was observed during the test. Subsequently, 2ml of benedict solution added to 2ml of extracted solution and no visible changes observed. Hence, there was presence of iodine and no presence of glucose or starch as shown in figure 3.

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Discussion From the cell simulation modal made, it has explained about the aspects of diffusion and indicator solution (chemical testing) to test for particular substances. From the observation made, iodine and glucose have greater concentration due to small molecular size and also they are monomers. It means their molecules are smaller. Thus, it diffused out to create a state of equilibrium. On the other hand, starch molecules did not move by diffusion across a membrane into or out of the cell due to the large molecular side. This is because many repeated glucose subunits are found in starch structure. Starch is a polymer. This is the reason why the water in the beaker or 1 inch dialysis tube did not turn dark blue as no starch molecules diffuse out. However, the liquid outside of cell was cloudy. When iodine indicator and Benedicts test, tested for presence of iodine and glucose respectively, the colour change was not very distinctive. Hence, we came to a rationale that more time could have needed for glucose and iodine achieve diffusion as 1 hour was not enough to complete as perhaps molecules on one side of a membrane become trapped by binding to macromolecule. On the other hand, perhaps water in the beaker could have been too diluted so could not detect the presence of glucose and iodine using indicators. This process is termed as osmosis. In osmosis, molecules also travel from higher concentration to lower concentration. However, osmosis only occurs in water. Water molecules are polar and small enough to pass through the membrane. Therefore, from this experiment, we can understand that water is essential for cells survival. However, too much of water will result in hypotonic. cells is placed with a lower concentration, then osmotic pressure of the extracellular fluid is less than intracellular fluid. As a result, water flow into the cell causing it to swell and undergo lysis (burst). Likewise, if a cell is placed into a solution with higher osmotic concentration than the intracellular fluid such as salt solution. The osmotic pressure of the extracellular fluid will exceed that of the intracellular fluid casing the cell to shrink and crenate. Hence, its hypertonic. This explains why when a small dose of fertilizer can enhance plant growth but over fertilization can kill the plant. Fertilization has salt and salt dehydrates the cytoplasm in the cell and would dry up making the cells shrivel up and die. Iodine indicators and Benedicts solution proved that when a substance reacts with its indicator solution, a reaction will occur and cause a colour change. In this case, iodine test used to test for the presence of iodine in the solution and Benedicts solution is use to detect glucose. It is important for Benedicts solution to be heated as its a way to prove that it is the mixing of the two that creates the bright orange colour. If the benedicts solutions were to be heated separately or the solution to be
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heated there will not be any results produced to test the presence of glucose. The indicator solution will react only if the substance is meant for is present, then it will react. Else there would not be any changes to observe. Indicators come in handy when dealing with unknown samples of ingredients to prove what a substance is. A negative control is used yield a negative result. This is to ensure that all the reagents are pure and there is no contamination that will give a false result. Conclusion In conclusion, we can conclude that diffusion occur spontaneously and molecular size and charges matters when molecule pass thought cell membrane unless its molecular size is small and its a monomer diffusion will occur spontaneously. However, if the molecular size is big and it might also hinders the passage way for smaller molecules to diffuse to a higher concentration to lower concentration to bring it to an equilibrium state. We also concluded that cell should maintain a isotonic environment to conduct its activity smoothly else in hypertonic or hypotonic, it will burst or shrivel and eventually kills the cell. Therefore, diffusion and osmosis are types of passive transport and happens spontaneously.

References
http://www.sfponline.org/uploads/336/DiffusionLabReport.pdf http://faculty.buffalostate.edu/wadswogj/courses/BIO211%20Page/lectures/lab%20pdf's/Diffusion %20lab%2006a.pdf http://www.studymode.com/essays/Diffusion-Through-A-Membrane-Lab-462261.html http://answers.yahoo.com/question/index?qid=20100615085037AAQJsXx http://www.indiana.edu/~nimsmsf/P215/p215notes/LabManual/Lab5.pdf

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