Compensation is a complex and very important part of flow cytometry and above all else, it must be done correctly. Fluorochromes emission spectra often "spill over" into other detector channels, thus creating a "false" electronic signals. Please see the following example figures:
In this example, the only marker the cells are labeled with is CD45 FITC. The reason we have a false signal in the PE channel is because of overlapping emission spectra. For more details on spectral emissions please see the Spectral Analysis Tools page and consult one of the tools on this page. Once proper compensation is in place, the following histograms should look similar to the ones shown below.
For additional information and details about compensation, please see one of the two tutorials below.
|Mario Roeder's Compensation Tutorial||
One of the best compensation tutorials on the web today.
|Salk Institute Compensation Tutorial||Another outstanding tutorial.|