Rat microglia isolation and characterization using multiparametric panel for flow cytometric analysis

Abstract

In the field of traumatic brain injury (TBI), characterization of microglia is traditionally done using imaging-based methods as immunohistochemistry (IHC), while flow cytometry (FC) is a supporting method that reassures the results. However, often, the flow cytometry method used, especially in the commonly used rat models, is not microglial specific, not comprehensive, and certainly not standardized. Here we describe the full, optimized procedure for isolating rat microglia, proper sample preparation for FC run and provide essential analysis guidelines. This protocol allows the specific identification of microglia, by discriminating them from other myeloid cells, with high-resolution observation of microglial subpopulations and their activation markers expression.