Is there a technique to measure the biofilm formation in the Hungate tubes of autotrophically growing anaerobes?
There should not be a difference in the method of detection for aerobes or anaerobes.
Generally biofilm measure was using Imaging and automated cell counting are the most common methods of biofilm quantification. Furthermore, the use of stains or fluorescent markers, in order to more accurately identify cells of interest and distinguish from culture debris, allow for easier and increased accuracy of cell counting and data interpretation. There are various methods to detect biofilm production like Tissue Culture Plate (TCP), Tube method (TM), Congo Red Agar method (CRA), bioluminescent assay, piezoelectric sensors, and fluorescent and light microscopic examination and please find the attached my recent publication.
I don't think being an anaerobe is a problem, as long as you provide the right conditions. Therefore, the standard method (where you stain the biofilm with crystal violet) would work. Did you try it?
Sally S Abulaila to grow the bacteria autotrophically I should apply gas, this can not work in our 96-well plate method
Sara Al Sbei No, I didn't mean the 96-well micro-titer plate method. The method implemented in our lab does not stick only to the 96-well plate. You can grow the bacterium of interest in tubes as well (under the conditions you want) and after the 24 h incubation, the washing steps won't affect the biofilm formed in any way.
An idea is to start with several triplicates, stop one triplicate at each measuring time, and then measure the OD for them. At end of the experiment, you will then have one triplicate for the final biofilm formed.
Since you are using tools designed for anaerobic culture, there should be no difference between aerobes and anaerobes detection methods.
Now it depends on what you want to study and measure, for example: if you want to measure cell biomass (use CV), cell metabolic activity (XTT) or cell viability (MTT) and measure Optical Density of them. For each study you must choose the necessary and adequate reagent for the studied cells.
Cordially.
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