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White Blood Cell Differential Count

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					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
        NEUTRO
        EOSINO
        BASO
        LYMPHO
        MONO
   BLOOD ROUTINE TEST
   INFORMATION :
   HEMOGLOBIN
   CELL PRODUCTION
   CELL FUNCTION
   HEMATOLOGIC+OTHER SYSTEM+PROGNOSIS
   RESPONSE+RECOVERY
     Process of blood routine test


1.   Specimen collection
    Venipuncture
    Capillary Puncture
2.   Hematology Automation
3.   WBC differential count
        Specimen collection



Blood container:
purple cap

Anticoagulant reagent:
EDTA-K2 1.5-2.0mg/ml
Hemocytometer
         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
too acidic suitable   too basic
      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
high HCT




           small angle

low HCT




           large angle
tail   body   head
        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.
        Microscopic exam

 10×  (low fold): overall smear quality,
  rouleaux, agglutination or parasites
 100× (oil Len): WBC Diff, RBC
  morphology
tail   body   head
                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
Morphology of WBC
 in peripheral blood
normal peripheral blood
        smear
Stab neutrophil

         Diameter:12-16

         Cytoplasma:pink

         Granules: primary
                   secondary

         Nucleus: dark purple blue
                  dense chromatin
Segmented neutrophil

           Diameter: 12-16

           Cytoplasma: pink

           Granules: primary
                     secondary

           Nucleus: dark purple blue
                     dense chromatin
                     2-5 lobes
 Eosinophil
 Diameter: 14-16
 Cytoplasma: full of granules

 Granules: large refractile
  orange-red
 Nucleus: blue

  dense chromatin
  2 lobes like a pair of glass
Basophil
 Diameter: 14-16
 Cytoplasma: pink

 Granules: dark blue –
  black obscure nucleus
 Nucleus: blue
Lymphocyte
    Diameter: small 7-9
                large 12-16

    Cytoplasma: medium blue

    Granules: small agranular
               large a few primary
               granules

    Nucleus: dark blue \round
              dense chromatin
Monocyte
 Diameter:   14-20
 Cytoplasma: grey blue

 Granules: dust-like
  lilac color granules
 Nucleus: blue

   large irregularly
  shaped and folded
Abnormal changes
    of WBC
  morphology
 Left-shift and right-shift of neutrophil:
   Left-shift: non-segmented neutrophil > 5%
   Right-shift: hypersegmented neutrophil>3%
Toxic Granulation
Auer Bodies(Auer Rod)
Hypersegmentation
Anisocytosis of neutrophil
vacuolization
Degeneration of nucleus
Dohle body

				
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posted:11/6/2011
language:English
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