Determination of total red blood Cells count

ByDeepak Rajput

Aim: Determination of total red blood corpuscles (RBC) count

REFERENCE:

1. Haematology, Practical Human Anatomy And Physiology, S.R. Kale et al., Nirali Prakashan, Eight Edition, 2002, pp. 9-16

Background:

Red blood cell count is an enumeration of red cells or is any erythrocyte count.
The red blood cells or erythrocytes are circular, biconcave, non-nucleated cells containing
hemoglobin and are embedded in blood plasma. After birth, bone marrow is the main site of
the formation of red blood corpuscles, which are involved in acting as a carrier of oxygen and
carbon dioxide. RBCs also maintain the ionic balance of the human physiological system and
maintain the viscosity of blood. Various pigments like bilirubin and biliverdin are derived from
RBC after their degradation. The basic principle is that the blood specimen is diluted (usually
200 times) with red cell diluting fluid, which does not remove the white blood cells but allows
the red cells to be counted under magnification in a known fluid volume. Finally, the number
of cells in undiluted blood is calculated and reported as the number of red cells/µl of whole
blood. Blood cell counts can be performed using the hemacytometer.

Significance: The red cell count is the number of red cells in one cubic millimeter of blood.
The normal values of the red blood cell count are:

Woman: 4-5.5 million per cubic millimeter
Men: 4.5-6.0 million per cubic millimeter.
Infants: 5- 6.5 million per cubic millimeter.

Variations in normal values are observed in pregnancy, severe burns, and diseased conditions,
and it also depends upon altitude. It drops below normal values in anemia and leukemia and
rises above those in polycythemia and dehydration. Therefore, the red cell count is useful in
diagnosis.

REQUIREMENTS: Neubauer chamber, RBC pipette, Coverslip, RBC diluting fluid, Needle,
spirit, cotton.

PROCEDURE:

  1. Sterilise the fingertip with a cotton plug soaked in spirit and let it dry.
  2. Take a bold prick with a needle to have a free blood flow and draw the blood in an RBC
    pipette up to 0.5 mark.
  3. Dip the RBC pipette in red blood cell diluting fluid and suck up diluting fluid up to the 101 mark.
  4. Rotate the pipette equally in your hands to mix the solution well by swirling.
  5. Take the hemocytometer and place it on the flat surface of the workbench. Place the cover slip on the counting chamber.
  6. Allow a small drop of diluted blood from the pipette to sweep into the counting chamber by capillary action. Ensure there is no air bubble, and the counting chamber must not be flooded.
  7. Leave the counting chamber on the bench for Three minutes to allow the cells to settle. Observe the cells by placing the counting chamber on the mechanical stage of the microscope.
  8. Focus on the center room of the chamber and start counting the cells from the upper left corner. It is advisable to complete all counts of the four squares and then move to the center square, the fifth square to be counted.

DATA ANALYSIS: No. of cells X Dilution factor X Depth factor X Total ruled area / Area
count

Where; Dilution factor = 200; Depth factor = 10; Total ruled area = 25; Area count = 5

RESULT: The number of red blood cells present in one µl of blood sample is___________

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Editorial Note

This article has been carefully researched and written by Deepak Rajput with a focus on accuracy, clarity, and evidence-based healthcare information. Pharmaacademias.com

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