TTC Staining of Rat Brain Tissue Slices: A Staining Procedure to Differentiate Viable and Necrotic Tissue in Tissue Slices After Ischemia-Reperfusion Injury
This video describes the 2,3,5-triphenyl-2H-tetrazolium chloride staining procedure to visualize rat brain tissue slices and differentiate viable and damaged tissue following middle cerebral artery occlusion and reperfusion.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Brain staining and slicing After the middle cerebral artery occlusion experiment, remove the brain, including the brainstem, from the skull, and wash it in ice-cold PBS. Choose the correct size of the brain stainless-steel matrix (see the Table of Materials) depending on the weight of the anim…
Representative Results
Figure 1: Matrices for the rat heart and brain slicing. (A) Rat heart, (B) rat brain.
Disclosures
The authors have nothing to disclose.
Materials
2,3,5-Triphenyltetrazolium chloride (TTC)
Sigma-Aldrich
298-96-4
Adult Rat Brain Slicer Matrix
Zivic Instruments
BSRAS001-1
Single Edge Razor Blades
Zivic Instruments
BLADE012.1
Sony Alpha a6000 Mirrorless Digital Camera
Sony
ILCE6000
Can be repalaced by any up-to-date digiatal camera
TTC Staining of Rat Brain Tissue Slices: A Staining Procedure to Differentiate Viable and Necrotic Tissue in Tissue Slices After Ischemia-Reperfusion Injury. J. Vis. Exp. (Pending Publication), e21046, doi: (2023).