B.Pharm Lab. Instruction Manuals

Pharmacology I

APHE Anatomy, Physiology, and Health Education

Pharmaceutical Analysis

Pharmacy study material

Bones and Skeleton System

Bone disease (Gout) (Rheumatoid arthritis) (Osteoarthritis) (Osteoporosis)

Cancer and music therapy

Memory of water

1.2.5 Anatomy, Physiology, and Health Education I Laboratory

Red blood cell

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

Normal ranges:

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 chamber.

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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