2.3.5-Triphenyltetrazolium chloride (TTC) is a lipid soluble photosensitive complex commonly used for detecting seed viability and detecting ischemic stem cells in mammalian tissues. It is the proton receptor of the pyridine nucleoside structural enzyme system in the respiratory chain, which can react with dehydrogenases in normal tissues and appear red. Dehydrogenases in viable seed embryo tissues and cells can reduce TTC to insoluble red stable triphenylformamide (TTF). If the embryo dies or its vitality declines, it cannot be stained or stained lightly. Therefore, the vitality of seeds can be identified based on the staining site and depth of the embryo.
Technical Parameter
1. Take the brain tissue sample to be tested (usually taken directly after anesthesia or after infusion with physiological saline), and after taking it out, freeze it at 20 ℃ for 20-30 minutes for easy slicing
2. Slice the brain tissue to be examined, with a typical animal sample layer thickness of 0.5-2mm
3. Slice into TTCStain (2%) and immerse in dark for 25-35 minutes
4. Slice and fix in 4% paraformaldehyde or 10% neutral formalin for 4-24 hours
5. Absorb clean water from the surface of the tissue, use image analysis systems such as IPP to measure the area of cerebral infarction and calculate the volume of cerebral infarction