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J Neurosci Methods. 2012 Jan 15;203(1):10-8. doi: 10.1016/j.jneumeth.2011.08.045. Epub 2011 Sep 3.

FACS purification of immunolabeled cell types from adult rat brain.

Author information

  • 1Behavioral Neuroscience Branch, IRP/NIDA/NIH/DHHS, 251 Bayview Boulevard, Baltimore, MD 21224, USA. danielle.guez@yale.edu

Abstract

Molecular analysis of brain tissue is greatly complicated by having many different classes of neurons and glia interspersed throughout the brain. Fluorescence-activated cell sorting (FACS) has been used to purify selected cell types from brain tissue. However, its use has been limited to brain tissue from embryos or transgenic mice with promoter-driven reporter genes. To overcome these limitations, we developed a FACS procedure for dissociating intact cell bodies from adult wild-type rat brains and sorting them using commercially available antibodies against intracellular and extracellular proteins. As an example, we isolated neurons using a NeuN antibody and confirmed their identity using microarray and real time PCR of mRNA from the sorted cells. Our FACS procedure allows rapid, high-throughput, quantitative assays of molecular alterations in identified cell types with widespread applications in neuroscience.

Published by Elsevier B.V.

PMID:
21911005
[PubMed - indexed for MEDLINE]
PMCID:
PMC3221768
Free PMC Article

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