Determination of rbc count by Pandian M

AIM
METHODS
PRINCIPLE
REQUIREMENTS
COMPOSITION
PROCEDURE
CALCULATION
NORMAL VALUES
PRECAUTIONS
CONDITIONS ALTER TOTAL RBCs COUNT
OBSERVATION/RESULT.

1. DETERMINATION OF RBC COUNT Pandian M DEPARTMENT OF PHYSIOLOGY DYPMCKOP

2. OBJECTIVE AIM METHODS PRINCIPLE REQUIREMENTS COMPOSITION PROCEDURE CALCULATION NORMAL VALUES PRECAUTIONS CONDITIONS ALTER TOTAL RBCs COUNT OBSERVATION/RESULT.

3. PRINCIPLE The blood is diluted with diluting fluid( hayem’s fluid ) (usually 200 times)with the diluting fluid, which doesn’t remove the WBCs, but allows the red cells to be counted in a known volume of fluid. Finally, the number of cells in undiluted blood is calculated and reported as the number of red cells per mm³ of whole blood.

4. Normal values In adult Males–4.5 to 6 million per cumm of blood Females—4.0 to 5.5 million per cumm of blood New borns—6 to 8 million per cumm of blood

5. RBC Diluting fluid Hayem’s fluid Composition Sodium chloride - 0.5g Sodium sulphate -2.5g Mercuric chloride -0.25g Distilled H2O -100ml Function of each components Composition Sodium chloride - maintains osmolarity Sodium sulphate -prevents aggregation of RBCs Mercuric chloride -acts as a preservative Distilled H2O -solvent

6. Other fluids… Dacie’s fluid Alternative to hayem’s fluid Composition Trisodium citrate – 3.13g Commercial formaldehyde (37% formalin)- 1.0ml Distilled water - 100ml Isotonic saline (?) Gower’s solution.

7. Different between over and under charge

8. x1 x2 x3 x4 x5

9. Actual image of RBCs after charge into chamber

10. Calculation Central Sq volume = 1mm x 1mm x 1/10 mm. Central Sq divided into 25 small Sq. therefore, dimension of each small Sq will be 1/5 mm x 1/5mm x 1/10mm. Find out the sum of all RBCs in corner 4 and central most Sq, say (X).= (x1+x2+x3+x4+x5) (X) no. of RBCs present in 5 small Sq. (X) no. of RBCs present in 1/5 x 1/5 x 1/10 x 5 cumm . (X) no. of RBCs present in 1/50mm ³. In 1/50mm³ X no. RBCs present. Therefore, in 1 cumm – 50 x (X) no of cells precent . Dilution is 1 :200. The RBC count in Undiluted blood is (X) x 50 x 200 million / cumm .

11. RULES WHILE COUNTING CELLS IN EACH SQUARE Count only the cells present inside each square Cells on the line – count only the cells that are there on the TOP and LEFT border Omit those on the RIGHT border & lower border This will avoid counting the same cell again

12. BLOOD COLLECTION Capillary/peripheral blood collection Things required: Disposable lancet, spirit, cotton Select the finger tip Clean the tip with spirit Allow it to dry Make a prick with the lancet Wipe the first drop – blood diluted with tissue fluid Allow the blood to flow freely- do not squeeze or milk the finger to get blood

13. METHOD OF DILUTION Take blood up to the 0.5 mark without any air bubble Take the dilution fluid up to the 101 mark Keep the pipette between the palms and mix the fluid The fluid in the bulb gets mixed completely 2 to 3 mins After u have to ignore few drops (from mark 1) and then charge into neubauer’s chamber.

14. CONDITIONS ALTER TOTAL RBCs COUNT Physiological Pathological

15. References MUHS Journal Text book of Practical Physiology G.K.Pal C.L.Ghai A.K.Jain