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MICROBIOLOGICAL ENGINEERING - QUESTION 28.2 : 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 occupied 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).
Question 88 - In the calculation of the growth of bacteria, colony forming unit (CFU) in serial dilution is used. In a laboratory, viable count assay is used to estimate CFU. Formula applied is CFU / mL = (number of colonies x dilution) / (amount plated, in unit mL). Acceptable plate count is either between 20 and 200 or between 30 and 300 according to 2 different references. A wastewater sample of 200 ml is added to and mixed with 1.8 L of sterile water. Another 200 ml of the mixture is added to and mixed with 1.8 L of sterile water. (a) Calculate the dilution of first mixture and the dilution of the second mixture. (b) 100 microlitres of wastewater samples from the first mixture and the second mixture are placed separately on 2 different alga plates. The first plate has 250 colonies and the second plate has 23 colonies. Calculate the average CFU / mL.
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