I have cover slip sized glass slide with some Au/Ag nano-particles spot having diameter of around 50 micro meter. I want to determine the viability of the Biofilm around that spot. How can I determine the viability of Biofilm without extracting Biofilm or bacterial cell from the glass slide.
You can perform an XTT assay - a colourimetric assay based on metabolic activity of the cells present.
You could also do confocal/fluoresence microscopy with a live/dead stain to see which/how many cells are live versus dead.
Dear Daniel,
I am considering live/dead staining but this will only give me information about membrane integrity. I wanted to perform celltiter blue or XTT assay but the problem is i do not want to analyse the complete cover slide, but only the area covered by NPs. I believe by any colourimeteric assay i have to check it for complete cover slide ( for e.g. in 6 well plate )
Hi Priyanka,
Indeed live/dead works on the basis of membrane integrity, but there are no other ways I know of to look specifically at viability other than in this manner for a specific area. You would stain for the whole biofilm, and visualise the area of interest compared with the area not covered. It's an indication of viability either by direct or indirect disruption of the membrane to kill the cells.
Have a look at promega who offer viability assays, but as far as I'm aware, it would be similar to that of an xtt, in that you'd be looking at the whole biofilm, not a specific area.
I also know no other ways Indeed live/dead works on the basis of membrane integrity.
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