For biofilm assay in gram negative bacteria, which approach should I follow?

Dear everyone, i would like ask about biofilm formation technique for Staphylococcus  pseudintermedius. As I did biofilm technique using the tryptic soy broth+ 1 percent of glucose in tissue culture plate. When I stained with crystal violet 0.1 percent and washed after ten minutes there is observably biofilm as known this bacteria formed biofilm, what is your suggestion?

Hi There,

I am completely new to research area of biofilm, I am wondering what is the best method to grow biofilm in the lab using E.coli (if possible). Secondly how can I track or monitor the growth of biofilm using any feasible and straight forward method.

Any reference or suggestion would be highly appreciated.

Thanks in advance,

Kind Regards,

Anil

If I am interested in disrupting the biofilm, why does it matter that my treatment would also affect the planktonic cells? Sure, it isn't technically a biofilm as it is found in nature, but neither is a biofilm that is growing in isolation from other microbes found in vivo either.

If I am to centrifuge my 96-well plate, for example, then wash my plate and apply treatment, wait 24h, repeat wash, and apply resazurin, what kind of lashback and criticisms would I receive from the biofilm community?

I have to prepare biotic surfaces to study adherence of the bacteria and the biofim formation on it. Can anyone recommend a simple method?

I tried to prepare biotic surfaces of epithelial cells from buccal cavity. According to the procedure, the cells dried, but the cells shrunk. Any suggestions?

We are studying biofilm formation on differently coated 15mm diameter uPVC discs - these are opaque and may be white or black in colour depending on coating. The current setup includes forming biofilms on discs, rinsing to remove non-adherent cells, fixing each disc to a glass slide and then adding a drop of LIVE/DEAD BacLight mixture (Invitrogen L7012) to the centre of the disc. After incubation, the excess stain is rinsed off, a coverslip is placed on top of the disc and the edges of the coverslip fixed to the underlying slide with nail polish. After drying, the whole piece is placed in the inverted microscope coverslip side down. When imaging it is possible to see a hue of green or red depending on filter but not to resolve cells, and it is unclear whether this is just autofluorescence of the disc or coating. The opacity of the discs seems to block much of the light.

Thank you for your assistance and advice.

I am trying to read a very thin biofilm on a glass substrate using the ATR, but I do not get any spectrum, but when I try to read the same thin biofilm on a substrate grown on stainless steel I get a considerably decent spectrum (although the intensity is very low considering the biofilm is very thin). My question is is this because the glass is IR transparent and steel is opaque, or is there more fundamentals why is this. Can the FTIR settings be adjusted accordingly. Please if anyone can advice me on this. Thanks.

I am currently investigating biofilm formation in Salmonella isolates using 96-well crystal violet assays. The protocol I follow utilizes multichannel pipettes (opposed to other rinsing and removal methods I have read about others using). From 1:100 diluted overnight broth plates are inoculated and incubated (at various temperatures), broths are removed after and wells are washed using MQH2O until clear. I then stain using 0.1% CV for approx. 15mins (using a higher volume than the broth; e.g. 100ul broth and 125ul 0.1% CV), then washing wells once more after. after drying for 48h or more I solubilize the dye using 30% acetic acid and record the OD600. when comparing the results to a blank negative control of 30% acetic acid (approx. 0.05 OD600) calculations are showing very high biofilm formation (high OD600 in contrast to the negative control) when the lab isolates are supposed to have for example low formation or even none. I suspect there is insufficient pipetting of planktonic cells/ CV leaving a coating which is skewing OD600 measurements. This issue is consistent across most replicates and temperatures. I would love to hear any insight, thank you.

I have previously used LB broth medium in biofilm inhibition experiments against P. aeruginosa. However, I could not detect an effective biofilm formation in the negative control wells. Which medium would you recommend for sufficient biofilm formation in P. aeruginosa?

Thank you.

Hey everybody, I need help with a project I am working on with biofilm and Ti02 nanoparticle antimicrobial activity.

I need to know what are the culture requirments for Strep vridians, and what biofilm assay can I utilize to measure both strep viridians and pseudomonas aeruginosa.

please let me know! Thank you!