Introduction

Multicolor flow cytometry is widely used to sort subpopulations of cells. Antibodies and/or viability markers are used to tag proteins as the basis for sorting. In multicolor flow cytometry, compensation is required because fluorescence from more than one fluorochrome may be detected due to the overlap between the emission spectra. Compensation is generally performed with single color compensation beads or cells. The WOLF Cell Sorter has two modes of compensation available to the user (automatic and manual). Here, we demonstrate the use of the WOLF Cell Sorter’s ability to compensate 3 colors using FITC, PE and PE-Cy5 labeled beads, and sort these beads after compensation.

Methods

Calibration beads from Spherotech were used for the compensation and bulk sorts. Beads were FITC, PE or ...

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