Do I need to do a live and dead dye on my sample to eliminate dead cells? What will happen if I don't?

The autofluorescence of dead cells will be very strong, and if dead cells are not excluded with dead-activated dye, clusters of cells distributed along the diagonal line may appear in the scatter plot of the analyzed results, which will affect the experimental results. Therefore, in order to ensure the accuracy of the experimental results, for tissue samples other than spleen, it is generally recommended to add dead-activated dye to exclude dead cells. In the case of blood samples, dead cells may also not be excluded if the sample is fresh and multiple staining of only a few cell surfaces is done, but this requires preexperimentation to determine that there are not many dead cells in the sample or that they will not interfere with the results.

The dead cell population of the available data can be analyzed to see if the distribution of these cells in the fluorescence channel and the corresponding proportions are essentially the same as the live cell population; if they are consistent, the dead and live cell staining can be dispensed with; if they are not, the addition of live and dead dyes is recommended.

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