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Osmosis, Filtration, and Active Transport

Introduction The plasma membrane of the cell helps allow it be selective about what may or not may not pass through. It allows nutrients to enter the cell and keeps unwanted substance s out. There are two types of cell transport: active and passive transport. In the experiments, both transports are used. Active transport means energy that is required while passive transport means energy that is not required. Two types of passive transport are Osmosis and Filtration. Osmosis is the diffusion of water through a semipermeable membrane and Filtration is the process by which water and solutes pass through a membrane from an area of higher hydrostatic pressure into an area of lower hydrostatic pressure. The main focus in these experiments are Osmosis, Filtration and Active Transport. In Activity 3, I hypothesize NaCl will show the most osmotic pressure since the water molecules will be able to pass through its membranes easier than the rest of the solutes. In Activity 4, I hypothesize that NaCl will have the most filtrate in its membranes than in any other solute due since filtration is form depends entirely on the pressure gradient. In Activity 5, I hypothesize that increasing the ATP in its solutes will also increase the amount of solute transported in NaCl. Material and Methods Activity 3: Simulating Osmotic Pressure Materials - PhysioEx 8.0 - Lab book - Textbook - Computer/Computer Program - Two Glass Beakers - Four Dialysis membranes: (20 MWCO), (50 MWCO), (100 MWCO), (200 MWCO) - Three Solutes: NaCl, Albumin, and Glucose - Membrane Holder - Membrane Dispenser - Solution Dispenser - Deionized water - Beaker Flush - Timer Method 1. Drag the 20 MWCO membrane between the two beakers. 2. Adjust the NaCl concentration of 8.00 mM in the left beaker and then click Dispense button. 3. Click Deionized Water under the right beaker and then click Dispense. 4. Set the timer to 60 minutes then click Start, to run the experiment. 5. At the end of the run, click Record Data button.

6. Click the membrane to return it to the membrane cabinet. 7. Repeat steps 1 through 5 with the 50, 100, and 200 MWCO membranes. 8. Perform the same experiment for Albumin and Glucose by repeating steps 1 through 6 for each solute. For Albumin, dispense 9.00 mM albumin in step 2 and for Glucose, dispense 10.00mM glucose in step 2. Activity 4: timulating Filtration Materials - PhysioEx 8.0 - Lab book - Textbook - Computer/Computer Program - Two Glass Beakers - Four Dialysis membranes: (20 MWCO), (50 MWCO), (100 MWCO), (200 MWCO) - Four Solutes: NaCl, Urea, Glucose, and Powered Charcoal - Membrane Holder - Membrane Dispenser - Memrane Residue Analysis - Solution Dispenser - Deionized water - Pressure Unit - Beaker Flush - Timer Method 1. Click and hold the mouse on 20 MWCO membrane, and drag it to the holder below the top beaker. Release the mouse button to lock the membrane into place. 2. Adjust the NaCl, urea, glucose, and powered charcoal windows to 5.00 mg/ml each and click Dispense. 3. Make sure the pressure unit on top of the beaker reads 50 mm Hg. 4. Set the timer for 60 minutes and then click Start. 5. At the end of the run, record the filtration rate and the amount of each solute present in the filtrate (mg/ml). 6. Now drag the 20 MWCO membrane to the holder in the Membrane Residue Analysis unit. Click Start Analysis to begin analysis of the membrane. Click on Record Data button to keep the data. 7. Click the 20 MWCO membrane again to automatically return it to the membranes cabinet and click Flush to prepare for the next run. 8. Repeat steps 1 through 7 three times using 50, 100 and 200 MWCO membranes. Activity 5: Simulating Active Transport Materials

- PhysioEx 8.0 - Lab book - Textbook - Computer/Computer Program - Two Glass Beakers - Four Dialysis membranes: (20 MWCO), (50 MWCO), (100 MWCO), (200 MWCO) - Three Solutes: NaCl, KCl, and Glucose - Membrane Holder - Membrane Dispenser - Memrane Builder - Solution Dispenser - Deionized Water - ATP Unit - Beaker Flush - Timer Method 1. In the Membrane Builder, adjust the number of glucose carriers and the number of sodium-potassium pumps to 500. 2. Click Build Membrane and then drag the membrane between the beakers. 3. Adjust the NaCl concentration to be delivered on the left beaker to 9.00mM, then click Dispense button. 4. Adjust KCl concentration to be delivered to the right beaker to 6.00 mM, then click Dispense. 5. Adjust the ATP Dispenser to 1.00 mM then click Dispense ATP. 6. Set the timer to 60 minutes and click Start. 7. Click Record Data after each run. 8. Click Flush on either side to clean both beakers. Repeat steps 3 through 6, adjusting the ATP concentration to 3.00 mM. 9. Click Flush to clean the beakers again. Repeat steps 1 through 6, dispensing 9.00 mM NaCl into the left beaker and 10.00 mM NaCl (replacing KCl) into the right beaker.

Results Activity 3 Results:


Osmosis Results
300 200 100 0 20 MWCO 50 MWCO 100 MWCO 200 MWCO

NaCl 272 0 0 0

Albumin 153 153 153 153

Glucose 170 170 170 0

NaCl is higher at 20 MWCO (272 Hg) while there is no Osmosis present in the other Dialysis Membranes. Albumin has Osmosis present in all the Dialysis Membranes (153 Hg) and Glucose did also except at 200 MWCO (0 Hg). Activity 4 Results:

Membrane Residue was present in all membranes. NaCl had the most filtration in all Dialysis Membranes except for 20 MWCO (0 mg/ml). Urea had two filtrations in 100 MWCO and 200 MWCO and didnt have any filtrations in 20 MWCO and 50 MWCO (0 mg/ml). Glucose had only one filtration in 200 MWCO and the rest was 0 mg/ml. Lastly, Powered Charcoal did not have any filtration in any of the dialysis membranes. Activity 5 Results:

Discussion The first experiment conducted, Simulating Osmotic Pressure (Osmosis) demonstrates how molecules can pass through the pores of most membranes. There are three different solutes: NaCl, Albumin, and Glucose and each with different pore sizes (20, 50, 100 and 200). The smallest pore size is 20 MWCO and largest pore size is 200 MWCO. The first solute tested was NaCl, the results on the table that Osmosis pressure did occur only on 20 MWCO and reached equilibrium at 272 Hg. This means that water molecules were able to pass through. However, the other water molecules were not able to pass through since the table shows it only reached 0g Hg. Albumin showed osmotic pressure in all membranes (20, 50, 100 and 200 MWCO) occurred and reached equilibrium with the same average osmotic rate of 153 Hg. Also, Glucose showed osmotic pressure in three of the four membranes (20, 50 and 100) and had the same average osmotic rate of 170 Hg. Glucose did not show any osmotic pressure in the membrane 200 MWCO since it did not reach equilibrium and the water molecules were not able to able to move between the sides of the membrane. The observed data showed that my hypothesis was incorrect when Albumin showed that it had the most osmotic pressure shown and reached equilibrium in all membranes when NaCl only showed one in stance that osmotic pressure was shown and reach equilibrium at 20 MWCO while the rest of the membranes showed no osmotic pressure. The second experiment conducted, Simulating Filtration helped explain how water and solutes can pass through a membrane from an area of higher fluid pressure into an area of lower fluid pressure. The table shows that as the MWCO increases, the filtration rate had also increased. Membrane residue was present in all membranes. The first solute tested was NaCl. In Filtrate for 20 MWCO was 0 ml/mg. The rest of the membranes (50, 100 and 200) all had the same average rate of fluid pressure which is 4.81 mg/ml. In Urea, the filtrate for membranes 20 and 50 had no fluid pressure present (0 mg/ml). However there was fluid pressure that was present in the 100 and 200 of the filtrate which both are 4.81 mg/ml. Glucose did not have any filtrates in 20, 50 and 100 but did have show fluid pressure was present in membrane 200 which was 4.39 mg/ml. Powdered Charcoal did not have any filtrates in all its membranes. The observed data shows that my hypothesis was correct about solute, NaCl having the most filtrate in its membranes than any other solute since filtration is the amount of fluids and solute form depends on the pressure gradient and on the size of the membrane pores. The last experiment conducted, Simulating Active Transport demonstrates how a cell expends cellular energy (ATP) to move substances across its membrane.