The simplest is morphological detection, Hoechst staining, observation with a fluorescence microscope, very cheap, more than 200 pieces of dyes bought, can not be used to death, plus if your apoptotic morphology is good, the images are beautiful The cycle is also short, and the results can be seen directly. There is also a DNAladder that everyone chooses. The steps are very simple. However, it may take a lot of effort to make a beautiful LADDER. There is also a flow cell to detect it. It is not expensive. Kinds should be used very much.
To determine whether there is apoptosis, Annexin v-PI can be double-stained, because this method is sensitive to early apoptosis. If no obvious apoptosis is detected, it may not be detected.
After measuring apoptosis with Annexin v-PI double staining, other methods can be used to verify according to the specimen or sample, but I don't think it is necessary. Many high-quality papers use only one method to measure apoptosis. Of course, you should also be based on your next experimental needs. If you want to further determine whether the apoptosis is the mitochondrial pathway or the death receptor pathway, you can detect the corresponding caspase or the kit for selecting the mitochondrial pathway.
Normal cells and early and mid-term apoptosis cells can be stained by Hoechst.
The Hoechst staining of normal nuclei is round, pale blue with deep blue particles inside;
The apoptotic cell nucleus is bright blue due to concentration, or the nucleus is lobulated and fragmented.
The morphological changes of nuclear chromatin during apoptosis are divided into three phases:
The nucleus of stage I is rippled or creased, and some chromatin is concentrated;
The chromatin of the stage IIa nuclei is highly coagulated and marginalized;
Phase IIb nuclear cleavage into fragments, producing apoptotic bodies
Shanghai Biotechnology Co., Ltd. is mainly engaged in apoptosis detection kits, human apoptosis inhibitory factor kits and other species, welcome to inquire.
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