1.2.5 Anatomy, Physiology, and Health Education I Laboratory
Lab 7 Erythrocyte count using improved Neubauer's chamber
Developed by Prova Biswas
Theory: Erythrocytes, also known as red blood corpuscles, contain the haemoglobin (Hb)
which carries the oxygen to cells and tissues, is responsible for the red colour of blood. The life span of normal RBC
is 120 days. The RBC is produced in red bone marrow in adult. Before birth, the production of RBC (erythropoiesis) first
occurs in the yolk sac of the embryo and later in the liver, spleen, thymus, and lymph nodes of a fetus up to seven months.
Thereafter, production of RBC occurs in red bone marrow. The agents, required for the synthesis of blood cells are called
haematinic agents (iron, folic acid, vitamin B12, erythropoietin, colony stimulating factors).
Fig 1. Steps of Erythropoiesis
Male: about 5.4 million per µl or cubic mm of blood
Female: about 4.8 million per µl or cubic mm of blood
Clinical significances: The decreased count of RBC indicates several conditions like
iron deficiency anaemia, megaloblastic anaemia, pernicious anaemia, thalassemia, sickle cell anaemia, aplastic anaemia,
chronic renal failure etc. Increased RBC production (Polycythemia vera) occurs in burn, lack of oxygen during rock
climbing, respiratory disorders etc., when the viscosity of the blood increases.
Materials: 0.9% w/v NaCl (normal saline), pipette, EDTA blood, Microscope, test tube, Neubauer's
Method: 1. Make a 1:200 Dilution using EDTA Whole blood and saline 2. Put one drop of
the solution in the Neubauer's chamber, using a pipette 3. Count the center square (R marked) of the chamber.
Calculation: Cells/ cubic mm of blood = Number of cell counted x dilution factor x 5 /depth of fluid (0.1)
i.e. 10,000 times no of cells counted.
Fig 2. Improved Neubauer's chamber
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