Dear all -
we would like to produce and study biofilms of Pseudomonas aeruginosa. We are not well equipped, only a simple microscope. I wonder if it could be possible to make estimations on the drug (peptide) activity on the biofilm by using a plate reader to grow the biofilm and somehow use this setup to follow the live and dead cell staining?
Thank you very much for your response!
Have a nice day
Kornelius
Hi Kornelius Zeth
Yes, this is actually a common and well developed approach. There are many variations that have been adapted but most involve growing biofilms in microtiter plates, washing and removing cell biomass, and then staining and quantifying biofilm biomass. This can be tricky with certain phenotypes of very thin/fragile pellicle biofilms, but you can scan the literature for variations of this assay to suit your specific needs.
https://app.jove.com/t/2437
Another very simple method that requires no specialized equipment is a glass-bead based assay. Glass beads can be sterilized easily and dropped on top of pellicle biofilms to measure their tensile strength. Alternatively, surface-attached biofilms can be grown on glass beads, then subjected to disinfectants and are then easily manipulated for downstream microscopy, viability experiments, etc.
Article The Bead Assay for Biofilms: A Quick, Easy and Robust Method...
Article Three biofilm types produced by a model pseudomonad are diff...
Best wishes with the research ahead!
ACA
Kornelius Zeth, have you heard of Congo red staining? Congo red stains only the the extracellular matrix (like biofilm and capsule) and does not stain the cells. Simply, plate the bacteria on LB+congo red, LB+congo red+your peptide antibiotic. You should be able to visulalize the difference in the biofilm formation capability of the bacteria in presence/absence of your drug. Use the drug in lower concentrations, so that you do not kill the bacteria and still observe the effect of the antibiotic on the biofilm formation capability. Hope it was helpful.
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