Evaluation of the number of positive cells from flow cytometric immunoassays by mathematical modeling of cellular autofluorescence

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Abstract
A method for automated evaluation of the percentage of positive cells from flow cytometric immunofluorescence histograms is presented. The method is based on a suitable mathematical representation of cellular autofluorescence distribution which is used to identify the negative cell distribution in the test histogram. An algorithm is developed and implemented in a computer program that quantifies the positive cells from the routinely available control and test histograms. Its accuracy is compared to that of the other currently used automated methods of analysis on the basis of a set of histograms with overlapping distributions and with known percentages of positive cells, generated by artificial mixing of real data.