A microplate spectrofluorometric assay for bacterial biofilms.
A spectrofluorometric assay was developed for quantification of bacterial biofilms grown on a microtiter plate. The method involved staining biofilms formed by gram-negative and gram-positive bacteria with wheat germ agglutinin-Alexa Fluor 488 conjugate, which selectively binds to N-acetylglucosamine residues in biofilms. The fluorescence of stained biofilms was measured with a fluorescent plate reader. This method was compared with a widely used microplate colorimetric assay involving crystal violet staining of biofilms formed by both gram-negative and gram-positive bacteria. A strong linear association existed between the two methods (r (2)=0.99/0.94). Being more sensitive and specific as compared to colorimetric method, the spectrofluorometric assay provides a better alternative for quantification and characterization of bacterial biofilms.
Burton
E
E
Kane Biotech Inc, Winnipeg, MB, Canada.
Yakandawala
N
N
LoVetri
K
K
Madhyastha
M S
MS
D000117:Acetylglucosamine / Q000737:chemistry; D018441:Biofilms / Q000254:growth & development*; D005456:Fluorescent Dyes / Q000737:chemistry; D006090:Gram-Negative Bacteria / Q000502:physiology*; D006094:Gram-Positive Bacteria / Q000502:physiology*; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence / Q000379:methods*
D005456:Fluorescent Dyes; D000117:Acetylglucosamine