In my experiment I am currently growing two strains of bacteria, and in order to test them in my experiment I have to grow the biofilms in 15 mL falcon tubes. I haven't yet found a great assay to determine cell proliferation. I have been running CV staining of biofilms then de-staining cells, and transfering that solution to plates to collect abs. I am open to any suggestions.
I was thinking about getting a OD 600 on the media that I pull off the tubes after 24 hr incubation, but was concerned that I wouldn't be able to quantify the biofilm growth. Thank you for any help you can give.
Dear Bryce, did you perform your biofilm experiment in flat bottom 96-well plates? You can obtain quantitative data by measuring both @600 nm bacterial growth and @595 nm after crystal violet stainig biofilm formation. Surface of falcon tubes or glass tubes can affact biofilm formation of your strains.
Dear Mert, thank you for your response to my question. I have been growing bacteria in both 96 well plates, and falcon tubes. I have been able to get OD600 results in the 96 well plates, but was looking for a way to quantify growth in falcon tubes. I was aware that glass is difficult for biofilms to form on, but haven't heard anything about falcon tubes. What affects the biofilms in falcon tubes? We've been using falcon tubes because one step in the experiment requires that the biofilms be lowered approx 10-12 cm below water. What tube would you recommend?
Dear Bryce, the attached paper could be helpfull. Both bacterial surface properties and material surface properties (ionic, hydrophobic, surface charge etc.) play role in biofilm formation. We carry out experiments with staphylococci on the basis of their biofilm formation on different surfaces and their surface properties (surface zeta potential, hydrophobicity, etc.). Surface roughness is another parameter. I think the surface of falcon tubes so smooth, this can affect biofilm formation. Maybe you can try to use polystyrene tubes.
Best regards.
Thank you for both of these papers they helped clarify the problem with using plastics. I really appreciate your help and knowledge! Thanks again! Mert Sudagidan
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