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  Section: Biotechnology Methods » Cell Biology and Genetics
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Estimation of Number of Erythrocytes [RBC] in Human Blood

Cell Biology and Genetics
  Cell Cycles
  Meiosis in Flower Buds of Allium Cepa-Acetocarmine Stain
  Meiosis in Grasshopper Testis (Poecilocerus Pictus)
  Mitosis in Onion Root Tip (Allium Cepa)
  Differential Staining of Blood
  Buccal Epithelial Smear and Barr Body
  Vital Staining of DNA and RNA in Paramecium
  Induction of Polyploidy
  Mounting of Genitalia in Drosophila Melanogaster
  Mounting of Genitalia in the Silk Moth Bombyx Mori
  Mounting of the Sex Comb in Drosophila Melanogaster
  Mounting of the Mouth Parts of the Mosquito
  Normal Human Karyotyping
  Black and White Film Development and Printing for Karyotype Analysis
  Study of Drumsticks in the Neutrophils of Females
  Study of the Malaria Parasite
  Vital Staining of DNA and RNA in Paramecium
  Sex-Linked Inheritance in Drosophila Melanogaster
  Preparation of Somatic Chromosomes from Rat Bone Marrow
  Chromosomal Aberrations
  Study of Phenocopy
  Study of Mendelian Traits
  Estimation of Number of Erythrocytes [RBC] in Human Blood
  Estimation of Number of Leucocytes (WBC) in Human Blood
  Culturing Techniques and Handling of Flies
  Life Cycle of the Mosquito (Culex Pipiens)
  Life Cycle of the Silkworm (Bombyx Mori)
  Vital Staining of Earthworm Ovary
  Culturing and Observation of Paramecium
  Culturing and Staining of E.coli (Gram’s Staining)
  Breeding Experiments in Drosophila Melanogaster
  Preparation of Salivary Gland Chromosomes
  Observation of Mutants in Drosophila Melanogaster
  ABO Blood Grouping and Rh Factor in Humans
  Determination of Blood Group and Rh Factor
  Demonstration of the Law of Independent Assortment
  Demonstration of Law of Segregation

To count the erythrocyte cells, or RBC, diluting fluid was used. Diluting fluid for RBC is an isotonic solution. This solution keeps the RBC cells unhemolyzed. The RBC were counted using an improved neubauer chamber, which has an area of 1 square millimeter and depth of 0.1 mm. It is made up of 400 boxes. The cells in the 4 corner boxes and 1 box in the middle were counted.

Reagents Required
  • EDTA: Ethylene diamine tetra acetic acid is used as an anticoagulant.
  • RBC diluting solution: This is prepared by dissolving 0.425 gms of NaCl in 50 mL of distilled water.
  • Syringe
  • Needle
  • Alcohol
  • Vials
  • RBC pipette
  • Cotton
  • Neubauer counting chamber
  • Cover glass
  • Microscope
Using a syringe, venous blood was drawn and poured into vials containing anticoagulant (EDTA) and mixed well. With the help of RBC, pipette blood was drawn up to the 0.5 mark. The tip of the pipette was cleaned with cotton and RBC diluting fluid was drawn up to the 101 mark. The pipette with blood and fluid was shaken well, avoiding bubbling, and kept aside for 5 minutes. The counting chamber was charged using the posture pipette. After charging the chamber, the cells were allowed to settle down to the bottom of the chamber. The chamber was placed on the stage of the microscope and using a 45X objective, the RBC cells in the smallest square were counted.

The normal range of RBC in an adult male is 5–6 million and in a female it is 4−5.05 million/mm3. In the case of polycythemia, the number of RBC increases. This disease is associated with heart disease. A low count of RBC results in anemia. Polycythemia may be pathologic due to the tumor in bone marrow, and the number of red cells may reach 11 million per cubic mm of blood.

The present determination of total RBC count shows about ____/mm3 of blood, which is nearly equal to normal range.


Number of cells in 1 mm3 of blood = Number of cells counted × dilution factor × depth factor / Area of chamber counted = Number of cells counted × 200 × 10
1/5 = Number of cells counted × 10,000


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