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Necrotic cells could be trypan blue positive. WST-1 Assay could reflect cell number by measuring mitochondrial dehydrogenase, therefore, you could not know how many cells die in dishes. When you define apoptosis in your experiment as caspase dependent cell death, measuring caspase activity is the best. PS Trypan Blue Staining actually is a method to see ...


Using different wavelength for reference give some advantages. You can measure a baseline of each well. All well might not have the same baseline because each well might not be the exactly same as others. In addition, you could make scratches on it, you might touch the bottom, or you get some dirt from your bench. And this can be corrected by A540 in your ...

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