I wonder what are the best options of live/death dyes for gram negative bacteria culture that can be done in a high throughput fashion.
we have used SytoxGreen and it seems to work quite well, at least with E. coli However, there are several ways that such dyes can be misleading. For examples, cells may be permeabilsed to the dyes but not actually dead. Also, antimicrobials that kill cells by binding cellular DNA will block dye signals, so that cells that are actually dead appear to be alive. It is more tricky than it appears, but certainly a dye approach is a sensibly idea for HT screening.
- Dear Hanh you may select on of the several dyes used in microbiology briefed at the following link. https://www.researchgate.net/publication/260433689_Labtop_for_Microbiology_Laboratory?ev=prf_pub
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we have used SytoxGreen and it seems to work quite well, at least with E. coli However, there are several ways that such dyes can be misleading. For examples, cells may be permeabilsed to the dyes but not actually dead. Also, antimicrobials that kill cells by binding cellular DNA will block dye signals, so that cells that are actually dead appear to be alive. It is more tricky than it appears, but certainly a dye approach is a sensibly idea for HT screening.
What is a dead bacteria, that's the question! You can use the Dibac for the loss of membrane potential, the Live/Dead Baclight kit (Syto9+PI) for the loss of membrane integrity, Ethidium Bromide for the efflux pump, FDA for the esterase activity...
Dear Hanh,
You can use Propidium Iodide dye for the analysis of bacterial cell viability.
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