Hi ,
I am planning to do cell proliferation assay and live/dead cell assay. I appreciate if you can share your experience. Why do we need to do both of assays to check cell viability?
Thank you.
MTT and LDH assays,( etc.)are measuring byproducts of cell death at multiple time points of your own choosing - they do not tell you exactly when cell death was completed, since most are done at either 24, 48 or 72 hours. Secondly, dyes leak out once the cell is lysed, leading to high background measurements. The easier and more accurate method to perform a live/dead cell assay is by using a system that collects live cell growth and death in real time. That way you can collect not only live/dead data, but kinetics of cell death, more accurate EC50s, and mechanism of action predictions based on shape of the growth/death curve over time, including DNA damaging agents or mitotic inhibition. When comparing proliferation between different cell lines, you can gather information on cell doubling time, adhesion strength, and kinetics of cell growth, or senescence. Yes, endpoint assays can be less expensive ( not always as in case of LDH, for example - kits costs money), but you are also limited in the amount of useful data from your assay.
Can use the IncuCyte® Cytotoxicity Assay uses the IncuCyte® Cytotox Reagents to make real time measurements of cell death based on cell membrane integrity, all within your incubator.
Hi Debby, I really appreciate your clear explanation. I am going to use PrestoBlue assay for cell proliferation.
I am new to cell culture work and I start gathering some knowledge on this. I would much appreciate if you prefer to share your email address for further contact.
Hi Manash,
Thank you for the answer. I will search about IncuCyte assay.
Since cell viability, survival and death determination or cell proliferation assays, respectively, do not show the same effects.
All processes can happen more or less independend of and/or depending on each other.
- Badges
- Science topic
- Similar topics
- Microbiology
- Assays