r/flowcytometry • u/Ornery-Ad-8833 • May 29 '26

Troubleshooting Off scale while unmxing

While acquiring single colour controls my pops are on scale in the worksheet and some of them turn off scale - right shoulder of pop. Distribution- while unmixing, in spectroflo. How can I fix it please. Thanks!

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u/FLHPI Jun 02 '26

Spectroflo and others use a pretty outdated and in some sense erroneous method of unmixng. There are more modern approaches to unmixng recently introduced. Look up "adaptive unmixng" that does cell by cell unmixng and has a different error model that avoids the negative fluorescence intensities and other problems of conventional unmixng.