GSM6704115: SingleMicroglialCells, NCvsHFD, rep4; Mus musculus; RNA-Seq
Mice were anesthetized and blood was collected by ventricular puncture. Mice were then perfused with phosphate-buffered saline (PBS). For microglia isolation, the hypothalamus, cerebellum, hippocampus, and cortex were dissected from the brain and manually dissociated in HBSS buffer. A cell suspension was prepared with a continuous 37% Percoll gradient at 2300 rpm for 30 min without brake, then cells were washed with 1x PBS and resuspended in FACS buffer and non-specific binding to Fc receptors was blocked by incubation with anti-CD16/32 antibody (BD Pharmingen, Cat# 553141) and the FACS antibodies BV711 conjugated anti-CD11b and Pacific blue conjugated anti-CD45 antibodies (Cat# 101241, Cat# 103126, BioLegend).  The CEL-Seq2 method was used for single cell sequencing pair-end multiplexing run, with 50-75 bp read length