MICROBIOLOGICAL ENGINEERING - QUESTION 28.3 : In the calculation of the growth of bacteria, absorbance, A in spectrophotometry is used. According to Beer-Lambert Law, A = e x l x c where A is the absorbance of the solution (no unit), l is the distance of light travels through the solution (in cm), e is the molar absorptivity or the molar extinction coefficient [ in L / (mol.cm) ]. For a particular solute and fixed path length : As / Ao = Cs / Co where Ao is the observed signal for a known concentration Co, and As is the observed signal for a sample concentration Cs. (a) For a cell concentration of 560 cells / mL, a spectrophotometre gives an absorbance reading of 1.0. A mixture of concentration 3600000 cells / mL can be diluted in several operations, with each operation having a dilution of 1:20. How many dilutions should be made so that the concentration of this mixture can be calculated within a range of A = 0.0 to 1.0. (b) In another experiment, a sample tube of 1 cm in width is used. Let A = 0.06 and e = 0.0012 ml / (cell.cm). Find the cell concentration of the sample.

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