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Protocol to isolate oligodendrocytes, microglia, endothelial cells, astrocytes, and neurons from a single mouse brain using magnetic-activated cell sorting

Samah HoumamiD, Dominika SiodłakiD, Kevin D. Pham, Casandra Salinas-SalinasiD, Sarah R. OcañasiD, Willard M. FreemaniD, Heather C. RiceiD

Abstract

The isolation of specific cell types of the brain is essential to study cell-type-specific differences in complex neurological diseases such as Alzheimers disease. This protocol isolates oligodendrocytes, microglia, endothelial cells, astrocytes, and neurons from a single mouse brain. The process involves gentle tissue homogenization, debris removal, and sequential sorting of five distinct cell types. We validate cell purity and viability using flow cytometry and RT-qPCR. This protocol is well-suited for a range of downstream applications, including genomics, transcriptomics, and proteomics. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=200 SRC="FIGDIR/small/666877v1_ufig1.gif" ALT="Figure 1"> View larger version (47K): org.highwire.dtl.DTLVardef@1426b97org.highwire.dtl.DTLVardef@1a5bb77org.highwire.dtl.DTLVardef@1b69f32org.highwire.dtl.DTLVardef@8da416_HPS_FORMAT_FIGEXP M_FIG C_FIG

PDFDOI

02 /Specification & Classification

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