Protocol for isolating and culturing microglia from the adult mouse brain using a magnetic-activated cell sorting system

Microglia are the resident macrophages of the central nervous system and are highly sensitive to environmental cues, making primary culture challenging. Here, we present a protocol for isolating microglia from adult mouse brains using magnetic-activated cell sorting (MACS). The protocol includes brain sample collection, enzymatic tissue dissociation, step-by-step MACS-based microglia isolation, and the establishment of primary microglial cultures. We validate isolated cells using flow cytometry and quantitative real-time PCR (real-time qPCR) to assess microglial purity.