Accurately correct for fluorochrome spillover in your flow cytometry experiments using our specialized online tool. This Flow Cytometry Compensation Calculator helps you achieve precise data analysis by accounting for spectral overlap. Simply input your uncompensated MFI and spillover percentages to get compensated results instantly.
Formula:
Flow cytometry compensation corrects for spectral overlap, known as spillover, where emission from one fluorochrome is detected in the filter meant for another. The fundamental principle is to subtract the contribution of spillover from the uncompensated signal.
For a simplified calculation of a compensated MFI in a specific channel (e.g., Channel X), considering spillover from another channel (e.g., Channel Y), the formula is:
Compensated MFI (Channel X) = Uncompensated MFI (Channel X) - ( (Spillover % from Y into X) / 100 × MFI of Channel Y )
- Uncompensated MFI (Channel X): The Mean Fluorescence Intensity measured in Channel X before any compensation.
- Spillover % from Y into X: The percentage of the signal from fluorochrome Y that bleeds into the detection channel for fluorochrome X.
- MFI of Channel Y: The Mean Fluorescence Intensity measured in Channel Y (representing the source of spillover).