Stereotactic Atlas-Guided Laser Capture Microdissection of Brain Regions Affected by Traumatic Injury

Abstract

The ability to isolate specific brain regions of interest can be impeded in tissue disassociation techniques that do not preserve their spatial distribution. Such techniques also potentially skew gene expression analysis because the process itself can alter expression patterns in individual cells. Here we describe a laser capture microdissection (LCM) method to selectively collect specific brain regions affected by traumatic brain injury (TBI) by using a modified Nissl (cresyl violet) staining protocol and the guidance of a rat brain atlas. LCM provides access to brain regions in their native positions and the ability to use anatomical landmarks for identification of each specific region. To this end, LCM has been used previously to examine brain region specific gene expression in TBI. This protocol allows examination of TBI-induced alterations in gene and microRNA expression in distinct brain areas within the same animal. The principles of this protocol can be amended and applied to a wide range of studies examining genomic expression in other disease and/or animal models.

Association between RMTg Neuropeptide Genes and Negative Effect during Alcohol Withdrawal in Mice

“…Laser capture microdissection (LCM) enables precise access to brain regions using anatomical markers for identification [ 58 ]. To ensure accurate dissection and high-quality RNA, our protocol included rapidly freezing the brain in liquid nitrogen then sectioning it into 30 μm thick coronal slices with a cryostat based on stereotaxic coordinates.…”