Absolute Eosinophil Count

Dr. Mohammed Abdul Aleem

Practical Pathology
Osmania Medical College

 The eosinophil is slightly larger than the

neutrophil, with a diameter of 1217 m.
The nucleus is usually bilobed but
occasional nuclei are trilobed.
Eosinophil granules are spherical; they
pack the cytoplasm and stain reddish
orange.
The cytoplasm is weakly basophilic,
ribosomes and rough ER are abundant.

Principle:
A sample of blood is diluted with a solution that selectively
stains the eosinophils and eliminates all other leukocytes
and erythrocytes from view.
Following mixing, the specimen is introduced into the
counting chamber and the number of eosinophils in a
known volume of blood is counted.

Blood Sample:
Direct capillary blood or EDTA anti-coagulated blood is used
for Absolute Eosinophil Count.

Method:
There are two methods:
1. Improved Neubauer chamber method
2. Fuchs-Rosenthal counting chamber method

Diluting fluid: For both methods, Dungers diluting fluid is used.

Composition:
1. Eosin 1% (aq) Solution 5 ml
2. Acetone 5 ml.
3. Distill water 100 ml.

Eosin stains the eosinophil granules bright red, water is used to lyse red
cells and acetone is meant for fixation of eosinophils.

Procedure
Suck anticoagulated blood up to mark 1 in WBC pipette
Wipe tip and outside of the pipette.
Draw diluting fluid up to mark 11 in the WBC pipette to get
dilution of 1:10.
Mix well by rotating the pipette.
Charge the Neubauers chamber after discarding 1-2 drops of
the mixture from the WBC pipette.
Allow the cells to settle down for a few minutes.
Count the eosinophils under low power (10x) in 4 large corner
squares.
Eosinophils are identified by having brightly red granules.

Calculations:
Number of eosinophils in 1 mm3 = (nx10)/4
(Dilution factor = 10)
Hence, Number of eosinophils per mm3 (l)= n 10
Number of eosinophils in 1 mm3 = n 25
(n is the total number of eosinophils counted in 4 corner
squares)

Fuchs-Rosenthal Chamber:
In this chamber, count the cells in entire 16 sq. mm area.
The depth of chamber is 0.2 mm.
Hence more cells are counted and gives more accurate
result.
Normal Range:
40-400 per microliter.

Eosinophil leucocytosis Eosinophilia

1. Allergic disorders:
i. Bronchial asthma
ii. Urticaria
iii. Hay fever
iv. Drug hypersensitivity
2. Parasitic infestations
i. Roundworm
ii. Hookworm
iii. Tapeworm
iv. Echinococcosis
3. Skin diseases
i. Pemphigus
ii. Dermatitis herpetiformis
iii. Erythema multiforme

4. Pulmonary diseases
i. Lefflers syndrome
ii. Tropical eosinophilia
5. Haematopoietic diseases
i. Chronic myeloid leukaemia
ii. Polycythaemia vera
iii. Hodgkins disease
iv. Pernicious anaemia
6. Miscellaneous conditions
i. Rheumatoid arthritis
ii. Polyarteritis nodosa
iii. Sarcoidosis
iv. Irradiation

Causes of Eosinopenia
Acute stress including trauma, surgery, burns, epileptiform
convulsions, acute infections, acute inflmmation, MI, anoxia
and exposure to cold
Cushing syndrome
Administration of various Drugs including corticosteroids and
ACTH, epinephrine (adrenaline) and other agonists,
histamine and aminophylline
Haemodialysis (during procedure)