Improved Compensation in Flow Cytometry

Conventional compensation of flow cytometry (FMC) data of an N-stained sample requires additional data sets, of N single-stained control samples, to estimate the spillover coefficients. Single-stained controls however are the least rigorous controls because any of the multi-stained controls are closer to the N-stained sample. We have developed a new, optimization-based, compensation method that is able to use not only single- but also multi-stained controls to improve estimates of the spillover coefficients. The method has been demonstrated on a data set from 5-stained dendritic cells (DCs) with 5 single-stained and 8 multi-stained controls. This approach is practical and leads to significant improvements in FCM compensation.

For further details about this method, please refer to the original publication by I.Sugár, J. González-Lergier, and Stuart C. Sealfon

Improved Compensation in Flow Cytometry by Multi-Variable Optimization
Cytometry, Part A, 2011

Download the compensation files referred to in the publication.