Excellent Dual Dye Kit to Check Mammalian Cell Viability

National Institute of Immunology, India
Vaccine Immunology Laboratory
Postdoctoral Researcher

Overall

Quality of Results

Ease-of-Optimization

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Company:

Thermo Fisher Scientific

Product Name:

Live/Dead cell viability/cytotoxicity kit

Catalog Number:

L3224

We used this to elucidate the signaling pathway involved in toxicity of natural toxins. During the study to standardize the dose of MAPK pathway inhibitors, we have used this kit to check the viability of IMR-32 cells before and after the treatment with toxin and inhibitors. The kit is easy to optimize and clearly distinguishes live and dead populations.

Experimental Design and Results Summary

Application

Fluorescence microscopy

Starting Material

IMR-32 cell line (can be used for any mammalian cell lines)

Protocol Overview

Culture the IMR-32 cells on coverslips or in chamber slides, at 70–80% confluency treat the cells with your agent and inhibitors. After completion of incubation time, wash the cells twice with PBS and stain the cells with 100–150 µL of dye solution diluted in PBS (20 µL of 2 mM EthD-1 stock solution and 5 µL of 4 mM calcein AM stock solution in 10 mL of D-PBS cell culture grade). Incubate the cells for 30–40 minutes in dark at room temperature and again wash with D-PBS and visualized cells under fluorescence microscope.

Tips

Wash the cells properly with D-PBS before staining with dye solution.

Results Summary

Clearly distinguished live and dead cell population in the treated samples. Live cells were visualized as green dots and dead cells as red dots with bright fluorescence.

DOI or PMID #

10.1007/s12035-014-8816-4

Additional Notes

None

Image Gallery

Summary

The Good

Easy to optimize and can be used at lesser concentrations mentioned in the protocol.

The Bad

None

The Bottom Line

Excellent Live/dead Viability/cytotoxicity kit to stain the mammalian cells within 30 minutes.

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