Red Blood Cell Count Test | Total RBC count | Procedure and Calculations | скачать в хорошем качестве

Red Blood Cell Count Test | Total RBC count | Procedure and Calculations |

Total RBC Count - This video explains the procedure of Red Blood Cell Count practical using Neubauer Chamber or Haemocytometer with calculations. It is also known as Red Blood Cell Count t in Neubauer Chamber. Red blood cell count by neubauer chamber For detailed information visit: https://www.ebiologylab.com/experimen... Principle of Red Blood cell count Test: The total number of RBCs per mm3 of blood is determined by diluting known volume of blood 200 times with fluid, which prevents hemolysis and coagulation. The blood cells are then counted in the volume of diluted blood in Neubauer’s counting chamber or Haemocytometer. The number of RBCS per mm3 of the undiluted blood is then determined by this method. Apparatus required: 1) Neubauer Blood Counting Chamber: https://amzn.to/2S9VpR2 2) RBC Pipette: https://amzn.to/3xutNoD 3) Red Blood Cell diluting fluid: (Hayem's fluid): https://amzn.to/3vFj75c https://amzn.to/3zAE0ln 4) Compound microscope: https://amzn.to/2U6Lj3S 5) 90% ethyl alcohol: https://amzn.to/3xsJpZS 6) Sterile cotton: https://amzn.to/3iOKdEa 7) Sterile needle: https://amzn.to/2U6LuMA 8) Reagent Bottles: https://amzn.to/3xvRBZj 1.) Neubauer’s hemocytometer: The instrument is consists of a special glass slide. The counting chamber is a very heavy thick glass slide usually at the center with three platforms, which is separated by wide grooves. The central platform is 0.1 mm lower than its neighbor. On the surface of the middle platform, there are two sets of ruled area. The area has 9 large squares, each has 1 square millimeter area and is elaborately ruled. The four corner squares are divided into 16 equal sized squares. The central square is ruled into 25 groups of 16 small squares, each group is separated by triple lines. The ruled surface area is 1/10 millimeter below the inner surface of the cover glass placed over the middle platform. Thus each smallest square of the corner has a volume of 1/4 x 1/4 x 1/10 or 1/160 mm3. Whereas the smallest square in the center has a volume of 1/20 x 1/20 x 1/10 = 1/4000 mm3. This central segment is used for Red Blood Cell counting. In four big squares, leukocytes or WBCs is are counted which are placed at the four corners of the erythrocyte counting Chambers. Red Blood Cell diluting fluid: (Hayem's fluid) Mercuric chloride(Disinfectant and prevents fungal and bacterial growth)- 0.25g + Sodium Chloride(maintains osmotic pressure and prevents haemolysis)- 0.5g + Sodium sulphate(prevents rouleaux i.e., formation of rows of cells and coagulation of blood) - 2.5g + Distilled water up to final volume 100ml. The procedure of Total Red Blood Cell Count Blood Test: 1. The counting chamber is placed on the microscope stage. The illumination is adjusted and the central groups of 25 squares for RBCs are focused. One can see all the squares in alone field. 2. 1 ml of Hayem's is taken in a watch glass. 3. Collection of blood: The tip of the finger is sterilized by 90% alcohol and allowed to dry. The finger is pr.cked with a needle to produce a drop of blood of adequate size 3 to 4 millimeter in diameter. The blood is sucked up to 0.5 mark into the Red Blood Cell pipette. 4. The blood is diluted with Hayem's fluid up to 101 mark by sucking the diluting fluid by the pipette kept in a watch glass. When the blood is taken up to the mark of 0.5 the blood is diluted only 200 times. The contents inside the bulb of the pipette is mixed for 3 to 4 minutes. 5. The first two drops of diluted blood from the pipette are discarded and then this mixture is run on the hemocytometer slide on both the chambers on both sides under a special coverslip. 6. The corpuscles are allowed to settle down for 3 to 4 minutes and then the number of Red Blood Cells are counted under high power of the microscope in the 5 counting areas. 7. The white cells are recognized by the refractile appearance given to them by the diluting fluid. 8. The cells are counted under high power lens. One may count the RBCs in 5 squares under low power and then under high power for comparing the results. The cells in four groups of 5 squares each i.e., a total of 80 squares. Red Blood Cell Count Calculation: Out of these 25 squares, we have counted the number of Red blood cells only from 5 boxes. Hence, total we have counted 16 x 5 = 80 squares. It is known that that 80 small squares have a volume of 1/50 mm cube. It is also known that cell count in 1 smallest square, which has volume of 1/4000 mm cube. Let "X" be the number of RBCs in 1/50 mm cub. of diluted blood. Cells in 1 mm cub. of diluted blood are "X" x 50. Dilution employed was 0.5(Blood):100(Diluting fluid) i.e. 200. Therefore, number of cells in 1 mm cub. of undiluted blood will be "X" x 50 x 200 = "X" x 10000 i.e. = 526 x 10000 RBCs = 5,260,000 / mmcube.