Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2018 Sep 18:(139):58513.
doi: 10.3791/58513.

Simultaneous Cryosectioning of Multiple Rodent Brains

Affiliations

Simultaneous Cryosectioning of Multiple Rodent Brains

Tabitha R F Green et al. J Vis Exp. .

Abstract

Histology and immunohistochemistry are routine methods of analysis to visualize microscopic anatomy and localize proteins within biological tissue. In neuroscience, as well as a plethora of other scientific fields, these techniques are used. Immunohistochemistry can be done on slide mounted tissue or free-floating sections. Preparing slide-mounted samples is a time intensive process. The following protocol for a technique, called the Megabrain, reduced the time taken to cryosection and mount brain tissue by up to 90% by combining multiple brains into a single frozen block. Furthermore, this technique reduced variability seen between staining rounds, in a large histochemical study. The current technique has been optimized for using rodent brain tissue in downstream immunohistochemical analyses; however, it can be applied to different scientific fields that use cryosectioning.

PubMed Disclaimer

References

    1. Lyck L, Dalmau I, Chemnitz J, Finsen B, Schroder HD. Immunohistochemical markers for quantitative studies of neurons and glia in human neocortex. Journal of Histochemistry and Cytochemistry. 2008;56(3):201–221. - PMC - PubMed
    1. Fritz P, Wu X, Tuczek H, Multhaupt H, Schwarzmann P. Quantitation in immunohistochemistry. A research method or a diagnostic tool in surgical pathology? Pathologica. 1995;87(3):300–309. - PubMed
    1. Walker RA. Quantification of immunohistochemistry--issues concerning methods, utility and semiquantitative assessment I. Histopathology. 2006;49(4):406–410. - PubMed
    1. Evilsizor MN, Ray-Jones HF, Lifshitz J, Ziebell J. Primer for immunohistochemistry on cryosectioned rat brain tissue: example staining for microglia and neurons. Journal of Visualized Experiments. 2015. - PMC - PubMed
    1. Harrison JL, et al. Resolvins AT-D1 and E1 differentially impact functional outcome, post-traumatic sleep, and microglial activation following diffuse brain injury in the mouse. Brain, Behavior, and Immunity. 2015;47:131–140. - PMC - PubMed

Publication types

LinkOut - more resources