It strikes fear into the hearts of new cytometrists. Compensation. More fights have started over the proper way to compensate at meetings than anything else. This article will strive to shed some light on the principles of compensation, and equip you with the tools necessary to achieve compensation mastery for your research experiments.
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Compensation is a critical topic for flow cytometry, yet it is poorly understood. Within a flow cytometer, the appropriate ranges of excitation and emission wavelengths are selected by bandpass filters. But when emission spectra overlap, fluorescence from more than one fluorochrome may be detected. To correct for this spectral overlap, the fluorescence compensation process ensures that the fluorescence detected in a particular detector derives from the fluorochrome that is being measured.
This article discusses the principles of compensation to help those who use flow cytometry to master it for research experiments.
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