A comparison of spectral unmixing to conventional compensation for the calculation of fluorochrome abundances from flow cytometric data

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.