It will be especially important to do this for the parameter you use to measure DNA content.Ĭonstraints fix certain aspects of the model, forcing the model to be exactly the same from file to file, if you apply a model to a group of samples. Debris can be removed by making a series of 2D gates on all combinations of parameters and removing the cells along the axes.There are now viability dyes available that can be used with cell cycle assays. Dead cells can be removed using a viability dye.Dots that are not along this diagonal have a different ratio, which is likely to be caused by agglomerates. These are cells with a consistent ratio between two measures of the forward scatter pulse. Doublets can be removed by plotting FSC-A versus FSC-H and gating on the cells along the most dense diagonal.Whatever steps you typically follow to do this can be applied to cell cycle data. Removing doublets, dead cells, and debris will improve the condition of data. This page contains suggestions for improving the fit of difficult to model cytometric data.
