Flow cytometry is extensively used for the immune-profiling of leukocytes in tissue during homeostasis and inflammation. The multiparametric power of using fluorescently conjugated antibodies for specific surface and activation markers provides a comprehensive profile of immune cells. This chapter describes the identification and characterization of myeloid populations using flow cytometric analysis in an acute model of resolving inflammation. This model allows the examination of heterogenic populations across different systemic and tissue locations. We describe tissue processing, antibody staining, and analysis, which include a newly described viSNE tool to generate two-dimensional clustering within myeloid populations. We also reference the use of transgenic reporter mice on specific myeloid cells that provides enhanced specificity and profiling when defining myeloid heterogeneity.
Methods in molecular biology (Clifton, N.J.)
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Kennedy Institute of Rheumatology, Oxford University, Oxford, UK. email@example.com.