The microtiter plate biofilm assay (see Basic Protocol 1) is a useful method for assessing bacterial attachment by measuring the staining of the adherent biomass. Because it utilizes a 96-well plate format, it is suitable as a tool for screening large numbers of bacterial strains or species.
What is bacterial biofilm formation?
Biofilm formation is a process whereby microorganisms irreversibly attach to and grow on a surface and produce extracellular polymers that facilitate attachment and matrix formation, resulting in an alteration in the phenotype of the organisms with respect to growth rate and gene transcription.
What is biofilm testing?
Biofilms are formed when free-swimming, planktonic microbes adhere to one another and to a surface, and are often resistant to antibiotics. MBEC assays can be used to determine the minimum concentration of a test compound that will penetrate and eradicate microbial biofilms.
How is biofilm formation measured?
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.
What is microplate method?
Microplate is a polystyrene plate consisting of 96 micro wells of either U- or V-shape. Grouping is carried out in micro wells. This method is sensitive and ideal for large number of samples (see Figure 787.1).
How do you know if you have biofilms?
What are the signs that a biofilm has developed? The wound that has been infected with bacteria forming a biofilm may be much slower to heal or not heal at all, and may not improve with standard antibiotics. It may look sloughy or have an unpleasant smell.
How can biofilm formation be prevented?
Chemical modifications are the main strategy for biofilm prevention on indwelling medical devices. Antibiotics, biocides, and ion coatings are commonly used chemical methods of biofilm prevention. They prevent biofilm formation by interfering with the attachment and expansion of immature biofilms.
What is a microplate used for?
The microplate is designed to measure absorbance in small volumes. It is typically used to determine the concentration of nucleic acids and proteins. It contains 16 spots which can be loaded with 2 microliters of different samples or replicates for subsequent absorbance measurements.
How is the microtiter plate assay used for biofilm formation?
However, a number of other colorimetric and metabolic stains have been reported for the quantification of biofilm formation using the microtiter plate assay. The ease, low cost and flexibility of the microtiter plate assay has made it a critical tool for the study of biofilms. Downloadvideo file. (24M, mov) Protocol 1. Growing a Biofilm
What is the microtiter dish assay used for?
The microtiter dish assay is an important tool for the study of the early stages in biofilm formation, and has been applied primarily for the study of bacterial biofilms, although this assay has also been used to study fungal biofilm formation.
Which is the best microtiter dish for biofilm formation?
Dilute the over night culture 1:100 into fresh medium for biofilm assays. A standard biofilm assay medium for P. aeruginosais M63 minimal medium supplemented with magnesium sulfate, glucose and casamino acids (see Table).
How is the extent of biofilm formation measured?
In this assay, the extent of biofilm formation is measured using the dye crystal violet (CV). However, a number of other colorimetric and metabolic stains have been reported for the quantification of biofilm formation using the microtiter plate assay.