Multicolor Panel Design Tips

Flow cytometry panel design overview

Always try to pair the brightest fluorochromes with the weakest expressing antigen. In case the expression level of an antigen is unknown, it is advisable to use brighter fluorochromes.

Avoid spillover or spectral overlap between fluorochromes by spreading them as much as possible across the spectrum.

Avoid fluorochromes that can be excited by more than one laser e.g. APC-Cy7.

Always include suitable compensation controls especially if points 3 and 4 above cannot be achieved.

It is also advisable to use FMO controls to gate differently stained sub-populations more accurately.

Consider using online multicolor panel designers provided by FACS technology companies.

A suitable 4-color panel to illustrate points 1-3. The color of the boxes correspond to the lasers that excite the respective fluorochromes.

Flow Cytometry Multicolor antibody panel

Keywords: FACS fluorophores, fluorochrome conjugates, multi color panel, flow validated antibody panel, excitation emission, FACS dyes

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