Question 89 - A hemocytometer is a device that is used for counting cells. In an engineering experiment, 100 microlitres of cell suspension is diluted with 50 microlitres of Trypan blue dye. Only death cells appear blue in color when stained with the dye. There are 57 cells detected in a hemocytometer, where 5.3 % of them appear blue when the chamber of the meter is placed under a microscope. Each square of a chamber can contain 0.0001 mL of liquid. (a) Calculate the number of viable cells. (b) The cells ocupied 5 squares. Calculate the average number of viable cells / square. (c) Calculate the dilution factor of the cell suspension by using the formula : Dilution = final volume / initial volume. (d) Calculate the concentration of viable cells / mL by using the formula : Concentration = (Average number of viable cells / square) x dilution x (square / volume).
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Answer 89 - (a) Number of death cells = Total cells x % of death cells / 100 = 57 x 5.3 / 100 = 3.021 rounded to 3 cells. Number of viable cells = Total cells - number of death cells = 57 - 3 = 54 cells. (b) Average number of viable cells / square = 54 / 5 = 10.8 cells / square. (c) Dilution = final volume / initial volume = (100 + 50) microlitres / 100 microlitres = 1.5. (d) Concentration = (Average number of viable cells / square) x dilution x (square / volume) = 10.8 cells x 1.5 x (1 / 0.0001 mL) = 162000 cells / mL. The answer is given by Kang Chuen Tat; PO Box 6263, Dandenong, Victoria VIC 3175, Australia; SMS +61405421706; chuentat@hotmail.com; http://kangchuentat.wordpress.com.
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