Single cell sequencing of microglia in developing white matter

Microglia were FACS-isolated from developing mouse corpus callosum at postnatal days 0, 7, and 21, then sequenced by 10X Genomics single-cell sequencing. C57Bl6J mice were anesthetised and perfused with phosphate buffered saline. Brains were extracted and placed on ice in HBSS with HEPES buffer. Corpus callosi were dissected out and homogenized in glass dounces and filtered. Following centrifugation, cells were resuspended in 10% fetal bovine serum and run through a Percoll gradient. Cells were collected in FACS buffer, spun, and resuspended in HBSS. Myeloid cells were sorted based on expression of Cd11b.