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Red blood cell count

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Red blood cell count

  1. 1. By: Shefaa Adel Hejazy. Umm Al-Qura University, Makkah. Faculty of Medical Sciences. Hematology Dept. 1st Semester 1433/2012
  2. 2. o RBC is a flexible cell, biconcave disc in shape, and around 8 µm in diameter. o The number of red cells per volume of blood, measured in microliters (µL) or cubic millimeters (mm3)
  3. 3. Screening: Follow up: -Health maintenance -Response to ttt -NSAIDs -Chronic Anemia To assess degree of .. Anemia and blood loss
  4. 4. Counting of RBCs can be performed either:  Automated Method: (Electronic hematology cell counter, e.g. COULTER) OR  Manual Method: (visual using a microscope and counting chmber), which is a cumbersome and less accurate.
  5. 5. o The solution used for red cell count is Isotonic with RBCs; doesn’t lyse leukocytes. o Leukocytes are normally too few which can be identified easily and won’t be interfered with erythrocytes count.
  6. 6. o Reagents and instruments: 1. Neubauer Chamber (Haemocytometer) & coverslips. 2. RBC diluting fluid/solution. Consists of 3.2 g of Na-citrate and 1.0 ml of formaldehyde solution made up to 100 ml with D.W. o Sample:
  7. 7. Improved Neubauer Chamber
  8. 8. o A thick glass slide with H shaped moats in it. o The area between two lines of H (center) is 0.1 mm in depth. o Moat prevents mixing of 2 samples on either side of chamber.
  9. 9. o Prepare a plastic tube (labeled). o Prepare 1:200 blood dilution (4 ml of diluent + 20 µl Blood). o So, add diluent to the tube. Mix the sample (5 times); then aspirate 20µl and transfer to the tube and mix. o Clean the Haemocytometer and coverslip with 70% ethanol followed by D.W &Leave to dry. o Place a coverslip on the Neubauer chamber.
  10. 10. o o Then, fill the chamber with the diluent (10 microliter) in each side. Leave chamber in humidity (petri-dish with wet filter paper) for 1-2 min. !? o Condenser slightly lowered. Iris diaphragm should be almost closed. o Place chamber on microscope stage. Start with 10X to focus; then with 40X count RBCs.
  11. 11. o RBCs should be counted in the central square of the chamber. o Select 5 small squares (One at each corner and one in the center).
  12. 12. W W RBC W W
  13. 13. 25 small Squares = 1 mm.sq W 3 mm.sq R
  14. 14. Central large square RBCs WBCs
  15. 15. Count all cells in specified squares, and multiply by the proper conversion factor; the number of cells per cubic millimeter can be determined.
  16. 16. o Count all cells within 16 squares and those lying on middle lines, EXCEPT …
  17. 17. RBCs (1012/L) = No. of RBCs counted X Dilution X 106 Volume (µl) o Dilution = 200 o Depth of the chamber = 0.1 mm. o Volume of 5 small squares = 0.02 µl So, Red cell count/ liter = N x 0.01 x 1012 i.e. RBCs= N x 1012/L
  18. 18. o Diluent should be correct. o No overflow in the moats. o No air bubbles and debris in the chamber area. o No scratches in the ruled area of the chamber. o Pipettes used must be clean and dry.
  19. 19. Age group RBC count Adult Male 4.7 – 6.1 X 1012 /L Adult Female 4.2 – 5.4 X 1012 /L Newborn 4.4 -5.8 X 1012 /L Infant/ children 3.8 – 5.5 X 1012 /L
  20. 20.  Decreased value ( RBCs) 1. Anaemia: due to Blood loss, production of cells, destruction of cells, and dietary insufficiency. 2. Diseases which affect the Bone Marrow such as: i. ii. iii. Leukaemia MM Hodgkin’s Lymphoma 1. Subcutaneous bacterial endocarditis 2. Rheumatic fever
  21. 21.  Increased value ( RBCs) 1. Polycythaemia vera 2. Secondary polycythaemia, e.g. smokers, high altitude, cyanotic heart defects, and COPD. 3. Dehydration 4. Acute poisoning 5. Severe diarrhea
  22. 22.