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Leukocyte Total and Differential Count

Leukocyte Total and Differential Count. Lab 5. -100x magnification Mean of 10 fields x 100 = WBC x 10 9 /L -400x magnification Mean of 10 fields x 1,500 = WBC x 10 9 /L. WBC TOTAL COUNT Can be determined: manually by haemocytometer and microscope

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Leukocyte Total and Differential Count

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  1. Leukocyte Total and Differential Count Lab 5

  2. -100x magnification Mean of 10 fields x 100 = WBC x 109/L -400x magnification Mean of 10 fields x 1,500 = WBCx 109/L • WBC TOTAL COUNT • Can be determined: • manually by haemocytometer and microscope • automated by haematological analysers • by estimation from a blood smear

  3. -Leukocyte estimation in a blood smear (400x): Normal number Increased Decreased

  4. DIFFERENTIAL LEUKOCYTE COUNT • Can be determined: • from a stained blood smear • automated by some haematological analysers, but these need prior validation

  5. DIFFERENTIAL COUNT IN A STAINED BLOOD SMEAR: BATTLESHIP METHOD

  6. How to perfom a differential count: • Count 100-200 leukocytes, note each cellular type identified • Avoid counting the same field twice • Do not focus on one section of the smear

  7. TYPES OF LEUKOCYTES • GRANULOCYTES: Neutrophils, Eosinophils, Basophils • MONOCYTES • - LYMPHOCYTES

  8. Normal neutrophils and a lymphocyte Neutrophils are similar in most domestic species, characterised by pale cytoplasm and dark nucleus segmented into 2-5 lobes, although granules are more readily seen in ruminants.

  9. Small and medium lymphocyte, cow Small lymphocytes, dog

  10. Normal neutrophil and monocyte Monocytes are similar in most species; large cells with basophilic cytoplasm (often vacuolated or with tiny pink granules), and a pale irregularly clefted or folded nucleus with clumped chromatin.

  11. Normal eosinophil and basophil Morphology of eosinophils varies widely with species; granules are large in horses, small & numerous in ruminants, accompanied by vacuoles in dogs, and rod-shaped in cats. Basophils have purple-black granules, are numerous in large animals and may be almost absent in healthy dogs and cats.

  12. Normal WBCs in different species, some examples (1) (2) Eosinophil Eosinophil and neutrophil (4) (3) Basophil Eosinophil and neutrophils

  13. REPORTING A DIFFERENTIAL COUNT By relative % of each WBC type (INCORRECT) Ways to report the values By absolute number of each WBC type (CORRECT) (%of each cell type x total WBC number)

  14. EXAMPLE • Dog nº1 : 70% segmented and 10.0 x 109/L WBCs • Dog nº2: 70% segmented and 1.0 x 109/L WBCs • Dog nº3 : 70% segmented and 50.0 x 109/L WBCs % Segmented neutrophils Absolute segmented neutrophils x 109/L • Dog nº1 : 70 (NORMAL) 7.0 NORMAL• Dog nº2: 70 (NORMAL) 0.7 (DECREASED) • Dog nº3 : 70 (NORMAL) 35.0 (INCREASED)

  15. INTERPRETING DIFFERENTIAL RESULTS Neutrophilia Eosinophilia Basophilia Lymphocytosis Monocytosis  in a particular leukocyte Suffix “penia” (i.e. Lymphopenia) in a particular leukocyte

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