A novel FACS-based method to isolate enriched populations of neurons, astrocytes, and microglia

We developed a novel FACS-based method to isolate enriched populations of neurons, astrocytes, and microglia. Whole mouse brains were digested, and populations of neurons, astrocytes, and microglia were subsequently enriched by FACS using a combination of positive gating on cell-type specific markers and stringent negative selection against markers of oligodendrocytes and endothelial cells. We confirmed a strong enrichment of corresponding cell-type specific genes in isolated populations and a reduction in gene markers of endothelial cells and oligodendrocytes, demonstrating that highly pure populations of neurons, astrocytes, and microglia were obtained. Overall design: CNS cells from untreated wildtype mice were isolated using fluorescent activated cell sorting.