Protocol to sequentially isolate mouse oligodendrocytes, microglia, endothelial cells, astrocytes, and neurons via magnetic cell sorting

Isolation of specific brain cell types is essential to study cell-type-specific differences in complex neurological diseases such as Alzheimer's disease. Here, we present a protocol for isolating oligodendrocytes, microglia, endothelial cells, astrocytes, and neurons from a single mouse brain. We describe steps for tissue homogenization, debris removal, and sequential sorting. We then detail procedures for validating cell purity and viability using flow cytometry and quantitative reverse-transcription PCR (RT-qPCR). This protocol is well suited for a range of downstream applications, including genomics, transcriptomics, and proteomics.