When setting up multicolor flow cytometry experiments proper compensation is extremely important since these experiments provide complex data. Compensation helps correct spectral overlap to match the various fluorophores used during cell staining, after which the data becomes easier to interpret. Compensation using cells for single color staining provides autofluorescence levels that are the same as those obtained during multicolor staining and is independent of the antibody host or isotype. However, valuable cellular material and antibodies targeting dimly expressed antigens or rare cellular populations create difficulties when using this approach. Furthermore, native cells are difficult to standardize and introduce additional variability. Compensation procedures using antibody capturing beads overcome some of these limitations. However, many bead kits are host specific and do not cover the full range of isotypes. In addition, high backgrounds upon violet laser or red laser excitation are observed for the vast majority of capture bead kits. As a result, Spherotech offers the COMPtrol line of antibody capture beads.
The COMptrol beads offer:
- low autofluorescence regardless of excitation wavelegth or detection bandpass
- and the enormous breadth of compatible hosts and isotypes makes COMPtrol capture beads a truly universal compensation tool.
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SpheroTechnical Note #18 will provide the following information: