TTC Staining of Rat Brain Tissue Slices: A Staining Procedure to Differentiate Viable and Necrotic Tissue in Tissue Slices After Ischemia-Reperfusion Injury
Published: April 30, 2023
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 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.
開示
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 |
| 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 |
