Understanding Unmixing Errors

Unmixing errors are a familiar and troublesome feature of spectral flow cytometry. They are often attributed to differences between the reference signatures measured from the single-color controls and the spectra observed in the experimental sample. This explanation is broadly correct, but it does not fully explain why some very small signature differences produce clear artifacts while others do not.

Please read our white paper Understanding Unmixing Errors in Spectral Flow Cytometry by our Principal Scientist, Nicholas Rohrbacker PhD. The paper reveals that the magnitude and visibility of an unmixing error are determined by the direction of the unmixing error, brightness of the signal, and overlap defined by the panel itself. This is contrary to current thinking of similarity as the defining characteristic of these properties.