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!

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