VC grows very quickly in LB. What you are likely seeing is an effect of quorum sensing. Once the cells grow to a certain density the QS molecules will accumulate and the downstream effect will be an induction of HapR and a subsequent repression of the biofilm genes. This causes the dispersal, not the washing steps (at least in my experience). You could try a few things: Use diluted LB (1:50 dilution, make sure you add NaCl back to 100%), try replacing the media (effectively getting rid of the QS molecules) every few hours, image at an earlier timepoint (6h, 12h, etc.), or use a flow cell system to grow the biofilms so that the media is continuously replaced and QS molecules never accumulate. My recommendation is the flow cell option but it also takes an investment and a lot of set up. Also not the easiest system to work with. You might be able to set up something like a drip-flow thing using conical tubes and such but I don't have experience with that.

Hi........

vbc grow LB medium good, an d another thing you use it instead of LB in zoberl marine broth with 2 % nacl.

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hi sir,

U can try with BHI agar with 1% glucose for the bio-film studies. brain heart infusion agar gives some what static bio-film on the coverslip for both static and dynamic conditions. i used this media, got best results.

Hello researchers,

Thank you for your responses. Much appreciated.