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White Blood Cell Differential Count

White Blood Cell Differential Count. Definition. The relative percentage of each type of white blood cells in peripheral blood. This experiment is a part of blood routine test. Person---5L BLOOD 2L CELL + 3L PLASMA RBC+WBC+PLT

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White Blood Cell Differential Count

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  1. White Blood Cell Differential Count

  2. Definition The relative percentage of each type of white blood cells in peripheral blood. This experiment is a part of blood routine test.

  3. Person---5L BLOOD • 2L CELL + 3L PLASMA • RBC+WBC+PLT NEUTRO EOSINO BASO LYMPHO MONO

  4. BLOOD ROUTINE TEST • INFORMATION : • HEMOGLOBIN • CELL PRODUCTION • CELL FUNCTION • HEMATOLOGIC+OTHER SYSTEM+PROGNOSIS • RESPONSE+RECOVERY

  5. Process of blood routine test • Specimen collection • Venipuncture • Capillary Puncture • Hematology Automation • WBC differential count

  6. Specimen collection Blood container: purple cap Anticoagulant reagent: EDTA-K2 1.5-2.0mg/ml

  7. Hemocytometer

  8. Principle of the experiment • Wright-Giemsa: a polychrome stain • Methanol : fixes cells to slide • methylene blue stains RNA,DNA ===Blue /purple • Eosin stains hemoglobin, eosin granules ===pink /red pH value of phosphate buffer is very important

  9. too acidic suitable too basic

  10. Procedure of the experiment • Methods of preparation • Spreader slide at 30-40 degree angle • Control thickness of the smear by changing the angle of spreader slide

  11. high HCT small angle low HCT large angle

  12. tail body head

  13. Stain of blood smear • Wright’s Giemsa stain: 1-2min • Phosphate buffer :15min • Wash with distilled water * The staining time depends on the concentration of the stain and room temperature.

  14. Microscopic exam • 10× (low fold): overall smear quality, rouleaux, agglutination or parasites • 100× (oil Len): WBC Diff, RBC morphology

  15. tail body head

  16. Observing direction: Observe one field and record the number of WBC according to the different type then turn to another field in the snake-liked direction *avoid repeat or miss some cells

  17. Morphology of WBC in peripheral blood

  18. normal peripheral blood smear

  19. Diameter:12-16 Cytoplasma:pink Granules: primary secondary Nucleus: dark purple blue dense chromatin Stab neutrophil

  20. Diameter: 12-16 Cytoplasma: pink Granules: primary secondary Nucleus: dark purple blue dense chromatin 2-5 lobes Segmented neutrophil

  21. Diameter: 14-16 Cytoplasma: full of granules Granules: large refractile orange-red Nucleus: blue dense chromatin 2 lobes like a pair of glass Eosinophil

  22. Diameter: 14-16 Cytoplasma: pink Granules: dark blue –black obscure nucleus Nucleus: blue Basophil

  23. Diameter: small 7-9 large 12-16 Cytoplasma: medium blue Granules: small agranular large a few primary granules Nucleus: dark blue \round dense chromatin Lymphocyte

  24. Diameter: 14-20 Cytoplasma: grey blue Granules: dust-like lilac color granules Nucleus: blue large irregularly shaped and folded Monocyte

  25. Abnormal changes of WBC morphology

  26. Left-shift and right-shift of neutrophil: • Left-shift: non-segmented neutrophil > 5% • Right-shift: hypersegmented neutrophil>3%

  27. Toxic Granulation

  28. Auer Bodies(Auer Rod)

  29. Hypersegmentation

  30. Anisocytosis of neutrophil

  31. vacuolization

  32. Degeneration of nucleus

  33. Dohle body

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