I believe you could use F4/80pos (mouse macrophage marker), CD3neg (remove T cells), CD19neg (remove B cells), CD11cneg (remove DC), CD49b (remove NK), Gr1neg (remove neutrophils and monocytes subcells).
F4/80+ cells from CD11b+ gated cells can serve as a direct staining panel. CD206 or CD68 can also be effectively used. Certain subpopulations of monocytes can also be F4/80 positive therefore i would suggest to include a monocyte marker (but not Ly6c or CCR2 since it can also be found on macrophages at low levels) to discriminate between monos and mqs. Good luck!
I agree that you should use a monocyte marker but F4/80 is not expressed by all splenic macrophages. Red pulp macrophages (F4/80++CD11b+ highly autofluorescent), marginal zone macrophages (CD209b+F4/80-), marginal zone metallophillic macrophages (CD169+F4/80-), plus monocytes and monocyte-derived macrophages (F4/80+CD11b+).
http://www.ncbi.nlm.nih.gov/pubmed/20133793
Thank you very much Andre Fernandes and Güneş Esendağlı. I'll talk to my boss what you guys have suggested.
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