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What is the difference between compensation and spectral unmixing?

Posted June 22, 2020


Both compensation and spectral unmixing are processes aiming to remove the interfering signals from other fluorescent molecules, which can usually be achieved automatically by appropriate software. Compensation uses the spillover matrix while spectral unmixing uses reference spectra for calculation, which are in fact mathematically similar. The main difference between these two methods is that spectral unmixing handles data from more detectors than compensation, and separates the individual spectral fingerprints rather than intensities at a specific wavelength.

Additional resources

Fundamentals of Flow Cytometry

Cell Meter™ FITC-Annexin V Binding Apoptosis Assay Kit *Optimized for Flow Cytometry*

Picot, J., Guerin, C. L., Le Van Kim, C., & Boulanger, C. M. (2012). Flow cytometry: retrospective, fundamentals and recent instrumentation. Cytotechnology, 64(2), 109-130.

Nolan, J. P., & Condello, D. (2013). Spectral flow cytometry. Current protocols in cytometry, 63(1), 1-27.