Immunohistochemistry of Mouse Brain Tissue Sections for Targeting Dopamine Neurons

Abstract

Source: Kummari, E., et. al., Laser Capture Microdissection - A Demonstration of the Isolation of Individual Dopamine Neurons and the Entire Ventral Tegmental Area., J. Vis. Exp. (2015)

In this video, we demonstrate the immunohistochemistry protocol for dopamine neurons in brain tissue sections for subsequent laser capture microdissection.

Protocol

All procedures involving animal samples have been reviewed and approved by the appropriate animal ethical review committee.

1. Preparation of Slides and Tissue

1. Use either Silane-prep slides or polyethylene napthalate (PEN) membrane glass slides.
2. For RNA isolation, dip slides in RNase decontamination solution, wash 3 times with RNase-free water then dehydrate with a graded series of RNase-free ethanol and vacuum dry for 30 min.
3. Cut tissue sections at 10 µm thickness using a cryostat and mount on the prepared RNase free slides. Keep the sections frozen during sectioning to preserve RNA quality. NOTE: Be sure that....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Silane prep-slides	Sigma-Aldrich, St. Louis, MO	S4651
95% Ethanol-RNase Free	Sigma-Aldrich, St. Louis, MO	E7148
100% Ethanol-RNase Free	Sigma-Aldrich, St. Louis, MO	E7023
Rnase Free Triton	Sigma-Aldrich, St. Louis, MO	T8787
Anti-Tyrosine hydroxyase antibody	EDM Millipore, Billerica, MA	Ab152
Alexa Fluor 488 goat anti-rabbit IgG	Life Technologies, Grand Island, NY	A-11008
RNaseZap	Life Technologies, Grand Island, NY	AM9780