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Purpose:
- to establish the percentage distribution of the
different leukocytes
Nucleated RBC under the Microscope
Equipment:
Microscope with oil immersion objective Corrected WBC Count
Stained slide
Formula:
Procedure: WBC count x 100
1. Under LPO, slect area where cells are evenly Corrected WBC count =
distributed 100 + number of NRBC counted
2. Place a drop of cedar wood oil and examine under oil
immersion lens.
3. Count 100 leukocytes in the smear. Microscopic view of 1 large square (Area for counting WBCs
under LPO)
Suitable and unsuitable regions for observation of stained
smear
Computation:
Cells counted in 4 large squares
WBC/cu mm =
of blood no of squares x depth of counting x dilution***
counted* chamber**
Small lymphocyte
- small lymphocytes are slightly
larger than erythrocytes (about
11 um in diameter and circular
in shape. Their cytoplasm is
scanty and clear blue, having no
granules. The nucleus stains
deeply due to condensation of
chromatin. In healthy adults,
30-45% of leukocytes are
classed as lymphocytes.
Large Lymphocyte
- large lymphocytes are round
and twice as large as the
erythrocyte (about 15 um in
diameter). The nucleus is large
and the cytoplasm plentiful.
The nuclear chromatin structure
and the coloring of the
cytoplasm are similar to those
of small lymphocytes, except
that a few azurophilic granules
are present in the cytoplasm.
Polymorphonuclear neutrophil
- the size, cytoplasm and
nuclear chromatin structure of
polymorphonuclear neutrophils
are similar to those of
neutrophilic band granulocytes.
However, the nucleus is
segmented into three lobes by 2
3. shake, discharge 3 drops, then fill the counting
filaments. In healthy adults,a
chamber
bout 50% of leukocytes are
4. count (10x objective) the cells of the 4 large corner
polymorphonuclear neutrophils.
squares (1mm each)
Most polymorphonuclear
5. calculate the leukocyte count on the basis of cells
counted, area counted and the dilution
neutrophils have tow or three
lobes.
Eosinophil
- eosinophils are slightly larger
than neutrophils. The cytoplasm
contains large orange-red
granules. The nuclear chromatin
structure is similar to that of
neutrophils but the nucleus is
only segmented into two
swollen oval lobes of equal size
by a filiform strand. In the
normal blood, about 3% of
leukocytes are eosinophils. The
level increases in cases of
parasitic diseases, such as
ancylostomiasis, allergic
reactions, such as bronchial
asthma, and chronic
granulocytic leukemia.
- fin -
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