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    Lab 1

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    joenica
    This experiment determines the fragility of red blood cell membranes when exposed to NaCl solutions of varying concentrations. Red blood cells placed in hypertonic solutions will shrink and may burst due to osmosis, while those in hypotonic solutions will swell and burst. The experiment subjects red blood cells to NaCl solutions from 0.85% to 0%, then measures the amount of haemolysis at each concentration. This osmotic fragility curve indicates the concentration of solutes inside red blood cells.

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    Lab 1

    Uploaded by

    joenica
    245 views4 pages
    This experiment determines the fragility of red blood cell membranes when exposed to NaCl solutions of varying concentrations. Red blood cells placed in hypertonic solutions will shrink and may burst due to osmosis, while those in hypotonic solutions will swell and burst. The experiment subjects red blood cells to NaCl solutions from 0.85% to 0%, then measures the amount of haemolysis at each concentration. This osmotic fragility curve indicates the concentration of solutes inside red blood cells.

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    This experiment determines the fragility of red blood cell membranes when exposed to NaCl solutions of varying concentrations. Red blood cells placed in hypertonic solutions will shrink and may burst due to osmosis, while those in hypotonic solutions will swell and burst. The experiment subjects red blood cells to NaCl solutions from 0.85% to 0%, then measures the amount of haemolysis at each concentration. This osmotic fragility curve indicates the concentration of solutes inside red blood cells.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as DOCX, PDF, TXT or read online from Scribd
    Download as docx, pdf, or txt
    245 views4 pages

    Lab 1

    Uploaded by

    joenica
    This experiment determines the fragility of red blood cell membranes when exposed to NaCl solutions of varying concentrations. Red blood cells placed in hypertonic solutions will shrink and may burst due to osmosis, while those in hypotonic solutions will swell and burst. The experiment subjects red blood cells to NaCl solutions from 0.85% to 0%, then measures the amount of haemolysis at each concentration. This osmotic fragility curve indicates the concentration of solutes inside red blood cells.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as DOCX, PDF, TXT or read online from Scribd
    Download as docx, pdf, or txt
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    LAB 1 : OSMOTIC FRAGILITY

    OBJECTIVES: To determine the fragility of the erythrocyte membrane against the haemolytic effect of NaCl solutions of varying concentrations

    INTRODUCTION: Osmosis is the diffusion of a solvent through a semipermeable membrane from a region of low solute concentration to a region of high solute concentration. The semipermeable membrane is permeable to the solvent, but not to the solute, resulting in a chemical potential difference across the membrane which drives the diffusion. That is, the solvent flows from the side of the membrane where the solution is weakest to the side where it is strongest, until the solution on both sides of the membrane is the same strength (that is, until the chemical potential is equal on both sides). Red blood cells are bound by a membrane which allows water to pass through while generally restricting the solutes. This process, called osmosis, causes cells to shrink due to loss of water when placed in hypertonic solutions. On the other hand, red cells absorb water (called endosmosis) when in a hypotonic medium. This results in swelling and ultimately haemolysis as the cell bursts. The osmotic fragility test uses this fact to determine the concentration of solute inside the cell by subjecting it to the haemolytic effects of solutions of different concentrations.

    MATERIALS: 11 test tubes, NaCl (0.85%, 0.75%, 0.65%, 0.55%, 0.50%, 0.40%, 0.35%, 0.20%, 0.10% and 0%), Distilled water, Blood, Pipette, Spectrophotometer and cuvettes and Centrifuge, PROCEDURE: 1. 11 test tubes prepared and labeled as follows: 0.85%, 0.75%, 0.65%, 0.55%, 0.50%, 0.40%, 0.35%, 0.20%, 0.10% and 0% NaCl. 2. 5 ml of NaCl pipetted in each test tube according to the label and 5 ml distilled water pipetted in test tube labeled 0% NaCl. 3. 10 L blood (with added heparin or sitrat which act as anti coagulant) was pipetted at each test tube. The content of the test tube mixed carefully and the test tube sentenced to resting period of 20 minute at room temperature. 4. Test tubes shaked carefully and centrifuged for 9 minute at the velocity of 2000 rpm. 5. Supernatant transferred to labeled cuvettes. 6. Using supernatant from the test tube which is labeled 0.85% NaCl as reference (0% haemolysis) OD of each supernatant calculated using spectrophotometer with the wavelength of 540nm. 7. Percentage of haemolyis calculated using the following formula: OD Sample X 100% OD of the sample with 100% lysis 8. OD recorded and percentage of NaCl converted to mole/L and mOsm/L. 9. A graph percentage of haemolysis vs. concentration of NaCl plotted. (This graph is known as osmotic fragility curve).

    QUESTIONS 1. A test tube with blood in it has a particular solution added to it. After several minutes, the solution is not clear anymore, but becomes red. Which solution was added to the blood to obtain this result? (a) 0.9 % saline (b) 5 % saline (c) Distilled water

    2. A 0.8% saline solution would be _____ to the cytosol of a cell (a) hypotonic (b) hypertonic (c) isotonic 3. If you placed a peeled apple or potato in a 5 % salt solution, it would: (a) gain weight (b) lose weight (c) stay the same weight

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