Dear scientific researchers,
I am doing confocal imaging experiments recently and want to stain live and dead cells, but I haven't found a particularly suitable dye yet. I know there are many live and dead cell dyes on the market, but I'm not sure which one is best for confocal imaging.
My requirements for dyes are: they can clearly distinguish live and dead cells under a confocal microscope, the background signal is as low as possible, and they can't have too much impact on the cell state, otherwise it may interfere with subsequent observations.
I wonder if anyone has used live and dead cell dyes that work particularly well and can recommend them? It would be best if you could share your experience, such as whether the staining operation is difficult, how stable the dye is, and how it actually works in confocal imaging. It would be even better if you could attach relevant references.
Thank you in advance!
Binbin Lee
We can provide Calcein/PI type live and dead cell staining kits. We recommend paying attention to MCE's HY-K1094 kit product (Viability/Cytotoxicity Assay Kit for Live & Dead Cells (Calcein/PI)).
This kit is suitable for confocal microscopy staining imaging, and its mechanism of action is:
- Calcein-AM: can penetrate the membrane of living cells, and is hydrolyzed by intracellular esterases into green fluorescent products (excitation/emission wavelengths of about 490/515 nm), which only accumulate and color in living cells.
- PI (propidium iodide): cannot penetrate the membrane of living cells, can only enter dead cells and bind to DNA, and exhibit red fluorescence (excitation/emission wavelengths of about 535/615 nm).
After optimization, this kit is suitable for confocal imaging scenarios, which can realize the fluorescence distinction and visualization analysis of live and dead cells.
For live/dead cell staining in confocal imaging, the LIVE/DEAD™ Viability/Cytotoxicity Kit (Thermo Fisher, L3224) is highly recommended due to its clear distinction between live and dead cells, low background, and minimal cytotoxicity. It uses Calcein-AM to stain live cells (green) based on intracellular esterase activity, and Ethidium homodimer-1 (EthD-1) to label dead cells (red) through compromised membranes. The staining is quick (15–30 minutes), requires no fixation, and is well-suited for high-resolution confocal imaging.
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