Abstract

Traditionally, flow cytometers acquired data using the same number of detectors as fluorochromes being measured in the experiment. More recently, spectral flow cytometers utilize a larger number of detectors than fluorochromes. This seemingly small difference opens the door to a wide variety of mathematical tools for the calculation of the true fluorochrome abundances from the raw detector values as compared with traditional compensation. This review will provide a brief overview of the mathematics and theory underlying traditional compensation and unmixing focusing on the differences between them and the additional information provided by unmixing approaches.

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