TTC Staining of Rat Brain Tissue Slices: A Staining Procedure to Differentiate Viable and Necrotic Tissue in Tissue Slices After Ischemia-Reperfusion Injury

Abstract

Source: Liepinsh, E., et al. Rodent Heart and Brain Tissue Preparation for Digital Macro Photography after Ischemia-reperfusion. J. Vis. Exp. (2022)

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

1. After the middle cerebral artery occlusion experiment, remove the brain, including the brainstem, from the skull, and wash it in ice-cold PBS.
2. Choose the correct size of the brain stainless-steel matrix (see the Table of Materials) depending on the weight of the animals (Figure 1B). Place the brain with its ventral side up in the brain matrix.
   NOTE: When seated in the matri....

Results

Figure 1: Matrices for the rat heart and brain slicing. (A) Rat heart, (B) rat brain.

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Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
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
Surgical blade	Heinz Herenz Hamburg Germany	BS2982
Weigh tray, 70 mL	Sarsted	71,99,23,212
Thermo-Shaker	BioSan	PST-60HL-4
Toothed tissue forceps	Agnthos	11021-12
Sodium chloride	Fisher bioreagents	BP358-10
Cover Glass Forceps, Angled	Fine Science Tools	11073-10