Examination of Erythrocytes | Massachusetts Institute of Technology - Edubirdie

University:
Massachusetts Institute of Technology
Course:
HST 198 | Health Sciences and Technology
Academic year:

2024
Views:

350

Pages:

2
Author:

mason mc44

EXAMINATION OF erythrocytes Practicum a. Tools and materials Materials: Capillary blood, cotton, 70% alcohol, Hayem solution, sterile lancet Equipment: Improved Neubauer Hemocytometer, Microscope b. Principle The blood will be thinned by the addition of Hayem's reagent, in an isotonic environment other than cells erythrocytes will lyse and can be easily counted under a microscope. c. Work procedures 1. Preparation of the counting booth a) Prepare the counting chamber and cover glass in a clean and dry condition b) Moisten with a little water the two embankments of the counting chamber c) Place the cover glass on top of the counting chamber then slide it up and down simultaneously repeat until Newton's rings (rainbows) are formed on both embankments. 2. Dilution of blood using an erythrocyte thoma pipette a) Suction blood until the limit mark is 101 b) Clean the outside of the pipette tip from any remaining blood that is still attached c) Suction the Hayem reagent to the 101 mark, avoiding any air bubbles d) Shake the thoma pipette for 2-3 minutes then discard the first 3-4 drops e) Put it in the counting chamber by pouring 1 drop on the edge cover glass f) Incubate for 2-3 minutes to give the cells a chance to spread and settle. 3. Counting Erythrocyte cells a) Count the erythrocytes under a microscope at 40x magnification. One particular error. What is often done is to count the number of erythrocytes using a small objective lens so the error becomes bigger. b) Count the erythrocytes in 16 small boxes, with a size of 0.05 mm x 0.05 mm and count erythrocytes in 5 medium boxes with a size of 0.2 mm x 0.2 mm. c) Cells that touch the left and top border lines Erythrocyte count Number of Erythrocytes ( /µL blood) = nx Fp Vb Vb = 80 x L x W x T = 80 x 1/20 x 1/20 x 0.1 = 0.02 µL blood Fp = 100 volume of erythrocytes taken n = Number of erythrocytes (red blood cells) counted in the counting room Fp = Dilution factor Vb = Calculated plane volume Normal values: Men 4.5 – 5.5 million/µL blood Women 4.0 – 5.0 million/µL blood

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