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2,3,5-Triphenyltetrazolium Chloride (TTC) is a fat-soluble photosensitive complex, commonly used to detect seed viability and ischemia-reperfusion injury in mammalian tissues. It acts as a proton acceptor in the pyridine-nucleotide enzyme system of the respiratory chain and reacts with dehydrogenases in normal tissues to exhibit a red color. Dehydrogenases in viable cells of seed embryo tissues can reduce TTC to insoluble, stable red triphenylformazan (TTF). If the embryo is dead or its viability is impaired, it cannot be stained or only lightly stained. Therefore, seed viability can be identified based on the staining location and intensity of the embryo.
Experimental Procedure for Brain Tissue Staining
| Step | Operation Details |
|---|---|
| 1 | Take the brain tissue sample to be tested (generally, the brain can be taken directly after anesthesia or after perfusion with normal saline). After removal, quick-freeze at -20℃ for 20~30 minutes to facilitate sectioning. |
| 2 | Section the brain tissue sample to be tested; the thickness of sections for animal samples is generally 0.5~2 mm. |
| 3 | Place the sections into TTC Stain (2%) and incubate in the dark for 25~35 minutes. |
| 4 | Transfer the sections into 4% paraformaldehyde or 10% neutral formalin for fixation for 4~24 hours. |
| 5 | Blot dry the water on the tissue surface, then use an image analysis system such as IPP to measure the cerebral infarction area and calculate the cerebral infarction volume. |
