rb cs count

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Red cell countRed cell count

MethodsMethodsA. Manual diluting method.

Principle:Using a diluting fluid that destruct all blood cells

leaving only RBCs intact.

With the aid of a hemocytometer, counts the number of cells expressed in millions per micro liter of blood.

ReagentsReagents

Gower’s solution: Sod. Sulphate, Glacial acetic acid and D.W

Hayem’s solution: mercuric chloride, NaCl, sod. Sulphate and

D.W. Physiological saline (0.85%)

EquipmentsEquipments::

HemocytometerCover slipPipettes

ProceduresProcedures::1. Dilution:

Add 2 ml of hayem’s solution to clean and dry tube.

Add 10 µl of the sample (whole anti-coagulated blood) and mix.

Dilution factor: 1:200

2. Charging

Charge the hemocytometer between the cover slip and the counting chamber.

Leave the cells to settle down.The depth factor is 1\10

3. Counting:

Count the RBCs in 5 medium squares (4 at the 4 corners & 1 in the center) inside the central large square.

Count those cells in L-shape manner

16 Tertiary squares

Reading (counting)Reading (counting)????? Cell /square.

CalculationCalculation::Erythrocytes / µl of blood= cells counted per

square х 10 (depth) х 5 х200(dilution)

ORErythrocytes / µl of blood= cells counted per

square х 10 (depth)х 25х200 (dilution)

= …………………. Cells / µl or cells / mm3

B. Other automatic B. Other automatic methodsmethods : :

Electronic cell counterFlow cell cytometers : depends

on laser lights.

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