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9 Differential

The document outlines the objectives and procedures for creating an ideal blood film, including the importance of differential leukocyte counts for diagnosing infections. It details the ideal characteristics of a blood film, common errors, and the clinical significance of various leukocyte types. Additionally, it provides information on equipment needed and methods for counting and analyzing white blood cells.

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mohawktagra
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22 views35 pages

9 Differential

The document outlines the objectives and procedures for creating an ideal blood film, including the importance of differential leukocyte counts for diagnosing infections. It details the ideal characteristics of a blood film, common errors, and the clinical significance of various leukocyte types. Additionally, it provides information on equipment needed and methods for counting and analyzing white blood cells.

Uploaded by

mohawktagra
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PDF, TXT or read online on Scribd
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Objectives:

1. To make ideal blood film.


2. To mention ideal character of blood film.
3. To recognize common errors of blood
film.
4. To fix and stain this film.
5. To determine the percent of each
category of leucocytes.
6. To mention the name of increase and
decrease of each type of leucocytes.
7. To mention some cases lead to increase
or decrease of each type of leucocytes.
 Determination of the total leukocytic
count is an important clinical
measurement, but a better diagnosis is
obtained by making a differential
leukocytic count. In a differential count
the percentage of each type of
leukocyte in the total W.B.Cs population
is determined . Each type of leukocyte
performs a different function against
infection, and each disease causes
different responses by the leukocytes.
Theory :
 making blood film in order to spread a
drop of blood on a slide to give thin (one)
layer of blood in order to count and
differentiate WBCs.
 The object of this experiment is to
determine the relative distribution of the
various types of W.B.Cs and to evaluate
the relative ratio or percent of each in the
whole blood .
Equipments :
 Microscope, slides, blood lancet, alcohol,
leishman’ s stain (the leishman powder is
dissolved as 0. 15 gm powder in 100 ml
methyl alcohol) or Geimsa stain (10%),
Cedar oil.
Procedures:
1. obtain a drop of blood by finger puncture. Place a
small drop on one end of a clean glass slide.
2. Hold a second slide (the spreader) at an angle of
45‫ ە‬to the last slide and move it toward the drop of
blood .allow the blood to spread along the edge of
the spreader slide, then move the spreader in a
smooth, fast motion to the other end of the first
slide.This motion will deposit a thin, evenly spread
film of blood across the slide. Allow the slide to air
dry.
3. Using dropper, cover the film with leishman s stain
(about 15 drops) and leave for 1-2 minutes during
which the methyl alcohol will fix the cells.
4. Dilute the stain on the slide by the same amount of
dist, water (15 drops) and leave for about 10-15
minutes during which the cells will be stained.
Blood film
Air dryness

Fixation by methyl alcohol staining

staining

Leishman’s 15%

Geimsa 10%
Ideal characters of blood film
1. Thick from one end and thin at the other
end with feathered tail.
2. Away from edges of the slide about 2 mm.
3. Long at least 2 cm .
4. Smooth even free from holes , streaks and
waves.
Errors in blood film
1. Jerky movement (wavy blood film).
2. Short blood film.
3. Very thick or very thin blood film.
4. Not thick at head.
LEUKOCYTES

(A) Granulocytes (B) Agranulocytes

Neutrophil ---- 55-70 % Lymphocyte ------- 23%

Eosinophil ---- 2%

Monocyte --------- 4%
Basophil ----0- 1%
5. Wash the film with distilled water, blot
gently with filter paper to dry the
bottom of the slide while the top is air
dried.
6. Examine the film, first under the low
power and then under oil immersion .
several techniqes are used:
a-Cross sectional method.
b-Battelment method.
c-Square method.
d-Straight edge method.
a-Cross sectional method.

b-Battelment method.

c-Square method.

d-Straight edge method.


May give false increase in neutrophils
(neutrophilia), as during spread of film
neutrophils tend to migrate tward the margin
and lymphocytes tend to condense in the center
The most preferable ones are (Cross
sectional method) and (Battelment
method).
7-Count the number of each type of
W.B.Cs on the slide recording each on a
tally counter 5-digits.
8-After counting l00 cells, express the
results in percentages.
Clinical significance
Neutrophilia = in pyogenic (suppurative) infection
as in acute appendicitis .
Basophilia = in allergic diseases an leukaemia .
Eosinophilia = in allergic & parasitic diseases,
scarlet fever .
Lymphocytosis = in choronic diseases as T.B .
Monocytosis = in malaria, healing processes and
chronic diseases .
Neutrophilic leukopenia = immunosuppression
protozool infection, mal nutrition & aplastic anaemia .
Lymphocytopenia = Administration of glucocorticoid
drugs .
Neutrophil lymphocyte ratio (N/ L ratio)

 In carnivora and human N/L ratio is 2 : 1


or 3: 1 while in bovine and poultry it
equals 1: 2.
 In camel the ratio is approximately 1:1.

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