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Total leukocyte count - TLC

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Total leukocyte count - TLC

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Total leukocyte count - TLC

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  1. 1. TOTAL LEUCOCYTE COUNT Dr. Shameera Begum
  2. 2. TOTAL LEUCOCYTE COUNT  Number of white blood cells in 1 µl or 1 cubic mm of blood  Normal range Age group Cells / cu.mm Adults 4000-11,000 At Birth 18,000 ± 8000 1 yr 6,000 - 15000 2 – 6 yr 5,000 - 15000 6-12yr 5000-13,000 Pregnancy Upto 15,000
  3. 3. LEUCOCYTOSIS TLC > 11,000/MM3 Physiological  At birth  Pregnancy  High temperature  Severe pain  Muscular exercise Pathological  Infections  Malignancy like leukemia  Severe haemorrhage
  4. 4. LEUCOPENIA TLC < 4000 CELLS /CU.MM  Bacterial infections(typhoid)  Viral infection (Hepatitis, Influenza)  Protozoal infection(Malaria)  Megaloblastic anemia  Bone marrow depression as in aplastic anemia, drugs, radiation
  5. 5. METHODS  Visual haemocytometer method  Electronic method
  6. 6. HEMOCYTOMETER  Hemo: blood  Cyto: cell  Meter: measurement/counter It is an instrument used to count the blood cells.
  7. 7. SPECIMENS  EDTA blood  Capillary blood
  8. 8. REQUIREMENTS  Microscope  Hemocytometer  WBC diluting fluid
  9. 9. It includes: a) Neubauer’s slide b) Cover slip c) RBC pipette d) WBC pipette
  10. 10. NEUBAUER’S SLIDE  It is the name given to a thick glass slide .  In the centre of the slide, there is an H- shaped groove.  On the two sides of the central horizontal bar, there are scales for counting the blood cells  The depth of the scales is 1/10mm or 0.1mm.
  11. 11. NEUBAUER’S CHAMBER  Neubauer’s slide with a cover slip over it, is called a Neubauer’s chamber  Four corner squares are meant for WBC counting.  Total = 64 small squares
  12. 12. WBC PIPETTE  It has a white bead  It has graduations upto mark 11
  13. 13. WBC (TURK’S) DILUTING FLUID: It is prepared as follows:  a) Glacial acetic acid: 2.0 ml  b) 1 % (w/v) gentian violet: 1.0 ml  c) Distilled water: 97 ml
  14. 14. PRINCIPLE  The glacial acetic acid lyses the red cells while the gentian violet slightly stains the nuclei of the leukocytes.  The blood specimen is diluted 1:20 in a WBC pipette with the diluting fluid.  The cells are counted under low power of the micro scope by using a counting chamber.  The number of cells in undiluted blood are reported per cu mm (µl) of whole blood
  15. 15. PROCEDURE  Draw blood up to 0.5 mark of a WBC pipette.  Carefully, wipe excess blood outside the pipette by using cotton.  Draw diluting fluid up to 11 mark.  Mix the contents in the pipette and after five minutes by discarding few drops, fill the counting chamber and allow the cells to settle for two to three minutes.
  16. 16. FOCUSING  4X to see the general formation of slide.  10X for WBC counting
  17. 17. COUNTING RULE  Do not count cells touching  Bottom line  Right line  This is to avoid double counting.
  18. 18. DILUTION FACTOR 0.5 part of blood is mixed in 10 parts of fluid So, 1 part of blood is in 20 parts of fluid Thus, dilution factor for WBC counting is 20.
  19. 19. CALCULATION OF THE VOLUME OF WBC SQUARES
  20. 20. No. of cells counted x dilution WBC count (per cubic mm) = ---------------------------------------- Volume of chamber N x 20 = --------------- 4 x 0.1 = N x 50 cells / cubic mm
  21. 21. ELETRONIC METHOD  Coulter – Automated haemanalyser Advantages Easy and rapid method Time saving Very large number of cells are counted rapidly High level of precision Disadvantages Costly Calibration error Nucleated RBCs are counted as leucocytes Platelet clumps counted as leucocytes
  22. 22. OTHER USES OF NEUBAUER CHAMBER  RBC count  Platelet count  CSF cell count  Sperm count
  23. 23. AUTOMATED HAEM ANALYZER
  24. 24. Thank You

Editor's Notes

  • other uses of neubauer , pictures
  • Description

    Total leukocyte count - TLC

    Transcript

    1. 1. TOTAL LEUCOCYTE COUNT Dr. Shameera Begum
    2. 2. TOTAL LEUCOCYTE COUNT  Number of white blood cells in 1 µl or 1 cubic mm of blood  Normal range Age group Cells / cu.mm Adults 4000-11,000 At Birth 18,000 ± 8000 1 yr 6,000 - 15000 2 – 6 yr 5,000 - 15000 6-12yr 5000-13,000 Pregnancy Upto 15,000
    3. 3. LEUCOCYTOSIS TLC > 11,000/MM3 Physiological  At birth  Pregnancy  High temperature  Severe pain  Muscular exercise Pathological  Infections  Malignancy like leukemia  Severe haemorrhage
    4. 4. LEUCOPENIA TLC < 4000 CELLS /CU.MM  Bacterial infections(typhoid)  Viral infection (Hepatitis, Influenza)  Protozoal infection(Malaria)  Megaloblastic anemia  Bone marrow depression as in aplastic anemia, drugs, radiation
    5. 5. METHODS  Visual haemocytometer method  Electronic method
    6. 6. HEMOCYTOMETER  Hemo: blood  Cyto: cell  Meter: measurement/counter It is an instrument used to count the blood cells.
    7. 7. SPECIMENS  EDTA blood  Capillary blood
    8. 8. REQUIREMENTS  Microscope  Hemocytometer  WBC diluting fluid
    9. 9. It includes: a) Neubauer’s slide b) Cover slip c) RBC pipette d) WBC pipette
    10. 10. NEUBAUER’S SLIDE  It is the name given to a thick glass slide .  In the centre of the slide, there is an H- shaped groove.  On the two sides of the central horizontal bar, there are scales for counting the blood cells  The depth of the scales is 1/10mm or 0.1mm.
    11. 11. NEUBAUER’S CHAMBER  Neubauer’s slide with a cover slip over it, is called a Neubauer’s chamber  Four corner squares are meant for WBC counting.  Total = 64 small squares
    12. 12. WBC PIPETTE  It has a white bead  It has graduations upto mark 11
    13. 13. WBC (TURK’S) DILUTING FLUID: It is prepared as follows:  a) Glacial acetic acid: 2.0 ml  b) 1 % (w/v) gentian violet: 1.0 ml  c) Distilled water: 97 ml
    14. 14. PRINCIPLE  The glacial acetic acid lyses the red cells while the gentian violet slightly stains the nuclei of the leukocytes.  The blood specimen is diluted 1:20 in a WBC pipette with the diluting fluid.  The cells are counted under low power of the micro scope by using a counting chamber.  The number of cells in undiluted blood are reported per cu mm (µl) of whole blood
    15. 15. PROCEDURE  Draw blood up to 0.5 mark of a WBC pipette.  Carefully, wipe excess blood outside the pipette by using cotton.  Draw diluting fluid up to 11 mark.  Mix the contents in the pipette and after five minutes by discarding few drops, fill the counting chamber and allow the cells to settle for two to three minutes.
    16. 16. FOCUSING  4X to see the general formation of slide.  10X for WBC counting
    17. 17. COUNTING RULE  Do not count cells touching  Bottom line  Right line  This is to avoid double counting.
    18. 18. DILUTION FACTOR 0.5 part of blood is mixed in 10 parts of fluid So, 1 part of blood is in 20 parts of fluid Thus, dilution factor for WBC counting is 20.
    19. 19. CALCULATION OF THE VOLUME OF WBC SQUARES
    20. 20. No. of cells counted x dilution WBC count (per cubic mm) = ---------------------------------------- Volume of chamber N x 20 = --------------- 4 x 0.1 = N x 50 cells / cubic mm
    21. 21. ELETRONIC METHOD  Coulter – Automated haemanalyser Advantages Easy and rapid method Time saving Very large number of cells are counted rapidly High level of precision Disadvantages Costly Calibration error Nucleated RBCs are counted as leucocytes Platelet clumps counted as leucocytes
    22. 22. OTHER USES OF NEUBAUER CHAMBER  RBC count  Platelet count  CSF cell count  Sperm count
    23. 23. AUTOMATED HAEM ANALYZER
    24. 24. Thank You

    Editor's Notes

  • other uses of neubauer , pictures
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