JoVE Encyclopedia of Experiments
Biological Techniques
0 views • 4:54 min • July 8th, 2025
On exposure to harmful environment conditions, a subset of bacteria enter a state of low metabolic activity, evading the detrimental effects. These bacteria, called persisters, can tolerate transient exposure to high concentrations of antibiotics.
To study the effect of osmolytes - small organic solutes - on bacterial persistence, first, obtain an exponential phase bacterial cell culture with pre-existing persisters below the limit of detection, suspended in media.
Next, transfer the suspension into osmolyte-containing wells of a microarray plate. Keep a set of wells without osmolyte as control. Seal the plate with a gas-permeable membrane to allow rapid gas diffusion for cell growth and incubate.
The osmolytes in the medium at appropriate concentrations induce osmotic stress, triggering some cells to reprogram their cellular metabolism from rapid to slow growth, forming persisters.
Now, transfer the osmolyte-treated and control cells to an antibiotic-containing persister assay plate. Seal the persister assay plate and incubate.
In the presence of antibiotics, normal cells undergo cell death, whereas persisters - being antibiotic-tolerant - survive. Last, collect a small fraction o
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