Flow Cytometry Panel Designer
Multi-laser excitation and emission spectral overlap validation with spillover compensation warnings.
How it works
Each fluorophore is matched to available laser lines and detector filters, and pairwise spectral overlap is computed from excitation and emission profiles. Combinations whose emission overlap would demand heavy compensation are flagged, since large compensation values inflate spread and degrade resolution of dim populations.
Frequently asked questions
How do I design a multicolour flow cytometry panel?
Assign your brightest fluorophores to the least abundant or most dimly expressed targets, and dim fluorophores to highly expressed markers. Spread antigens across different lasers where possible to minimise spillover, avoid pairing fluorophores with heavily overlapping emission on the same laser, and always include single-stain compensation controls plus fluorescence-minus-one controls for any gate you intend to draw.
What is spillover and why does compensation matter?
Spillover is signal from one fluorophore detected in another fluorophore’s channel, because emission spectra are broad and overlap. Compensation mathematically subtracts that contribution. It corrects the median position of a population but cannot remove the added measurement spread, so a heavily compensated pair produces wider distributions and can obscure genuinely dim populations.