I have used the following markers in a flow cytometry panel to assess mouse samples from bone marrow, spleen, fat pad, peyer’s patch and lymph node:
Ly6G
F4/80
CD11b
CD11c
SiglecF
FceR1a
I selected the granulocyte population in the fsc-a vs ssc-a plot. Then I gated on the viable cells and then on the singlets. To identify neutrophils I gated on fsc-a and Ly6G. Then I took the neutrophil population and divided it between siglecf and Ly6G, and labeled the double positives eosinophils. Is this correct? If so, can I use the same strategy for all the organs?
I also divided the granulocyte population between cd11c and cd11b. I then took the cd11c- and cd11b+ population and divided it between cd11b+ and f4/80. Is this the right way to gate on macrophages?
What else can I do with these markers using what gating?
Thank you!