I want to test for tumor-associated macrophages, is it okay to use only the F4/80 test?

The teacher is advised to add CD45, CD11b and dead-viable dyes. On the one hand, the large variety of cells and the low percentage of macrophages in tumor samples make it difficult to find target populations. On the other hand, dead cells are easy to take up antibodies and probes, resulting in obvious non-specific staining, and dead cell autofluorescence is particularly strong, which will lead to enhanced background fluorescence, preventing us from observing weakly positive expression of some markers. Therefore, it is recommended that the teacher exclude dead cell interference by dead and live dyes, then circle CD45 positivity to find the target cell population, and finally determine the macrophage ratio by CD11b and F4/80.

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