I worked on antibiofilm activity of some enzymes by crystal violet assay on 96- wells , but what's the best way to destaining CV 95% ethanol or 33% glacial acetic acid and why ?
95% ethanol is much better than GAA. Gram positive cell wall has a stronger attraction for crystal violet when Gram's iodine is applied than does the Gram negative cell wall. It is able to form a complex with the crystal violet that is attached more tightly to the Gram-positive cell wall than to the Gram-negative cell wall. This complex can easily be washed away from the Gram-negative cell wall with ethanol. In the case of GAA, over-destaining has been observed. That's why ethanol is more preferred.
Hi all , i worked with klebsiella biofilm (gram negative ),but according to your experiments , ethanol 95% is better than GAA?? but if we compain them (ethanol and GAA ) can we have a better results ?
Dr.subbaiah has contributed significantly in Hydrometallurgy.Sevral plants have come up because of his effort.
Are you trying to measure biofilm mass (or cell mass in biofilm) by using crystal violet staining? I'm not fond of that method for several reasons. I've used a fluorescence-based substrate and published on it, and easy to use if you have a microplate reader and microplate washer.
I am working with anti-biofilm activity assay. Please support me a optimal protocol. Thanks very much!
Again, need to identify if you are looking for a measure of residual bacteria from a biofilm or a measure of the organic matrix that comprises a biofilm? Which is it?
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