int asst practical

1. 1 WBC Count Specific Equipment 1. White pipette 2. Rubber pipe 3. Plastic syringe 4. Pipette vibrator 5. Hemacytometer & cover slip (Counting chamber) 6. Microscope 7. Counter (hand tally) 8. Report form

2. WBC Count Reagent WBC diluting fluids 1. Turk’s solution Glacial acetic acid Gential violet Distilled water 2. Two % acetic acid 3. One % hydrochloric acid

3. 3 WBC Count Specimens 1. Fresh blood 2. Anticoagulated blood EDTA Double Oxalate Citrate Heparin Etc.

4. WBC Count Procedure 1. Mix the blood sample thoroughly. 2. Fill blood into white pipette at 0.5 mark. 3. Fill reagent add up into the pipette to 11 mark. 4. Shake the pipette on the vibrator for 1 min. 5. Discarded the first 3-4 drops. 6. Fill in the hemacytometer nicely. 7. Allow WBC to set down for 2-3 min. 8. Count 4 white squares under microscope 9. Calculate the WBC concentration.

5. 5 WBC Count 1 mm. W W 1 mm. R R 1 mm. R R R W W High 0.1 mm. 3 mm.

6. 6 WBC Count W 1 mm. High 0.1 mm. 1mm.

7. 7 WBC Count Calculation 1. Volume of 1 white square = 1 x 1 x 0.1 cu.mm. = 0.1 cu.mm. 2. Volume of 4 white square = 0.1 x 4 cu.mm. = 0.4 cu.mm. 3. In 0.4 cu.mm. the WBC count = N (counted No.) 4. In 1 cu.mm. the WBC count = N x 1 / 0.4 = N x 2.5 5. The dilution for WBC = 0.5 / 10 = 20 6. The final WBC count = N x 2.5 x 20 = 50 N (/cu.mm.)

8. 8 WBC Count Calculation White cell count = number of cells counted (N) x volume factor (=2.5) x dilution factor (=20) = N x 50 Normal range = 5,000 - 10,000 / cu.mm. 5 - 10 x 103 / cu.mm.