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Differential Leukocyte Count

This document provides instructions for performing a manual differential leukocyte count. Key steps include: 1. Making a well-stained thin blood film on a clean slide using fresh blood without anticoagulant or EDTA anticoagulated venous blood. 2. Examining the smear under the microscope using different magnifications and counting at least 100 leukocytes using a classification method. 3. Reporting the results as percentages of different types of leukocytes and calculating their absolute counts using a provided formula involving the total white blood cell count. 4. Reference values for the relative and absolute counts of neutrophils, lymphocytes, monocytes, eosinophils, and basophils are also listed

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Mariam Khalil
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384 views2 pages

Differential Leukocyte Count

This document provides instructions for performing a manual differential leukocyte count. Key steps include: 1. Making a well-stained thin blood film on a clean slide using fresh blood without anticoagulant or EDTA anticoagulated venous blood. 2. Examining the smear under the microscope using different magnifications and counting at least 100 leukocytes using a classification method. 3. Reporting the results as percentages of different types of leukocytes and calculating their absolute counts using a provided formula involving the total white blood cell count. 4. Reference values for the relative and absolute counts of neutrophils, lymphocytes, monocytes, eosinophils, and basophils are also listed

Uploaded by

Mariam Khalil
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Manual differential leukocyte count

Introduction:
A differential white cell count is the assigning of leucocytes to their individual categories, this
categorization being expressed as a percentage or, when the WBC is available, as an absolute
count..

Materials:
1- Clean, dust- free, grease-free glass slide (better to use glass slide with frosted end for
labeling).
2- Spreader (smooth end).
3- Fresh blood without anticoagulant or EDTA anticoagulated venous blood. (Do not use
heparinized venous blood because it leads to blue back ground).
4- Leishman’s stain

Methodology:
1- Make well-made well stained thin blood film.
2- Examine the smear systematically by using X10,X40 and X100.
3- Start counting in an ideal area (not more than 50% of RBCs are overlapped).
4- Classify at least 100 leukocytes either by battlement or longitudinal method.
5- Report results of the 100 cells classified as percentage.
6- Calculate the absolute leukocyte count by using the following formula:

Absolute leukocyte count= TWBCs count X relative count

100

1
Reference values:

Absolute count Relative count

Neutrophils 2.0–7.0 × 109/l (40–75%)

Lymphocytes 1.0–3.0 × 109/l (20–45%)

Monocytes 0.2–1.0 × 109/l (2–10%)

Eosinophils 0.02–0.5 × 109/l (2–6%)

Basophils 0.02–0.1 × 109/l (<1–2%)

*WBC Correction for the Presence of Nucleated Red Blood Cells (nRBCs)

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