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Counting BHK Cells using a Coulter Counter

The Beckman Coulter Particle Counter was used to determine the concentration of BHK cells harvested in a test tube. 200 uL of cells were diluted in 20 mL of Isoflow. A 20 mL Blank was also created. Both the Blank and the Sample was counted 3 times. The data was averaged to account for any discrepancies in a single run. The data was Normalized by subtracting the average Blank away from the average Sample. This value was divided by 0.5 mL, as this was the volume drawn into the Coulter Counter at every run. This alone would indicate the diluted concentration in the cup. To determine the cell concentration in the test tube, the diluted concentration must be multiplied by the Dilution Factor. As we had 0.2 mL of cells in a total solution of 20.2 mL, the Dilution Factor was 101. Finally the machine was cleaned for the next person to use.