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Perform RBC Count

The document outlines the procedure for counting red blood cells (RBCs) using the Improved Neubauer chamber method, including the necessary materials and compositions of diluting fluids (Hayem’s and Dacie’s). It details the step-by-step process for preparing the blood sample, charging the chamber, and counting the RBCs under a microscope. Additionally, it provides the normal RBC count ranges for different demographics and discusses clinical significance related to increased or decreased RBC counts.

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ai6094810
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7 views4 pages

Perform RBC Count

The document outlines the procedure for counting red blood cells (RBCs) using the Improved Neubauer chamber method, including the necessary materials and compositions of diluting fluids (Hayem’s and Dacie’s). It details the step-by-step process for preparing the blood sample, charging the chamber, and counting the RBCs under a microscope. Additionally, it provides the normal RBC count ranges for different demographics and discusses clinical significance related to increased or decreased RBC counts.

Uploaded by

ai6094810
Copyright
© All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PDF, TXT or read online on Scribd

Aim: To perform red blood cells (RBCs) count by Improved Neubauer chamber method

Principle: The whole blood is diluted with an isotonic solution and the Erythrocytes/Red
blood cells in a portion of the diluted fluid are counted in a haemocytometer under a
microscope.
Requirement
1. RBC diluting fluid: Hayem’s or Dacie’s fluid
2. Improved Neubauer counting chamber
3. Red cell pipette
4. Cover slip
5. Tissue paper

Composition of Hayem’s fluid


Constituents Quantity Action

Mercuric chloride 0.5g  Acts as a fixative and


preservative
 Prevent growth of
bacteria

Sodium chloride 1.0g  Maintains isotonicity


 Prevents swelling or
shrinkage of RBCs

Sodium sulphate 5.0g  Maintains cell


separation
 Prevents clumping of
the RBCs
Distilled water 200ml  Acts as a solvent

Composition of Dacie’s fluid


Constituents Quantity Action

Sodium citrate dihydrate 5.0g  Acts as a


anticoagulant
 Prevents RBCs
clumping
Formalin 1ml  Acts as a fixative and
preservative
 Preserves cell shape
and prevent
hemolysis
Distilled water 100ml  Acts as a solvent

Procedure
1. Gently mix the blood in the EDTA vial
2. Draw the blood up to 0.5mark of RBC pipette. Wipe off excess blood from the sides
of the tip of the pipette
3. Dip the tip of the pipette in the RBC diluting fluid and the fluid up to mark 101.
4. Holding the pipette horizontally in its long axis, rotate it slowly to ensure thorough
mixing of the blood and diluent.
5. Place the cover slip on the ruled area of the chamber
6. Discard first 2 to 3 drops of the RBC fluid from the pipette
7. Charge the chamber by placing the tip of the pipette just under the cover slip and
allow the fluid to flow by capillary action till the counting chamber is just filled.
8. Wait for 2-5 minutes for the cells to settle in the chamber
9. Examine the central ruled area for red cell counting by examining under low power
objectives
10. Count RBCs using high power objectives in the 5 red squares and total them
together.
Calculation
RBC count = No. of cells counted(N) X Dilution Factor/ Area X depth
Where:
• Dilution factor =200
• Area of 5 square =1/5
• Depth of the chamber = 0.1mm
RBC count= NX200/(1/5X0.1)
RBC count = N X 10,000 million /cu mm
Normal range of the RBC count
1. Adult male: 4.4-6.0 million/cu mm
2. Adult female: 3.8-5.0 million/ cu mm
3. At birth: 6.0-7.0 million/cu mm
Clinical significance:
Causes of decreased RBC count
 Anaemia
 Haemodilution
Causes of increased RBC count
 Polycythemia
FIG: RBCs pipette

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