I have stored biofilms before for later study. I found if the biofilms stay in stagnant liquid for extended periods there is a build-up of free planktonic cells and a lack of replenished biofilm cells. As you know, biofilms develop best when there is a constant flow of new media over them such as in a flow cell. If you are growing them in well plates I would recommend using only freshly grown biofilms for your studies to ensure accuracy of results.

Yes, it is possible, but the way you do it depends on the (future) intended use. Also you can save the biofilms, per se, or biofilm fractions (for example, matrix, matrix components, etc...)

I think there is no point to store the biofilm for longer period for two resons

1-if you store it fresh it just keep growing and the results will be changed

2-if you store it in stain Gram stain or Live/Dead stain it will effect your results

Unless you will sperate the biofilm compound such as EPS etc in this case is easy to store in -80

Hope my comment is helpful :)

This depends upon what parameter or model you are testing. Biofilms grow at different rates, under different conditions and with different nutrient requirements. It is best to examine the biofilm promptly instead of storing it. Biofilms are complicated because as soon as you attempt to freeze the instant of the experiment you are modifying the results of the experiments. Practically, I have snap frozen biofilms (liquid nitrogen), scraped off cells and added glycerol and frozen at -80C. These experiments were variously for 16S rRNA typing, protein analysis, recreation of specific biofilms. In order to examine biofilm inhibition over a longer time period I have recently used slow growing marine organisms at low temperatures (10C), dilute broth concentrations and no agitation to form visible biofilm (8-10 days).

It would be better to test on fresh biofilms instead of storing. When you do store then they still grow and the conditions becomes different.

Hi Karthikeyan,

You can use the same biofilms continously. Maybe you can check this simple protocol:

Article Synthetic DNA molecules in biosensing of biofilms

Good luck,