A microplate spectrofluorometric assay for bacterial biofilms
A microplate spectrofluorometric assay for bacterial biofilms
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DOI：
10.1007/s10295-006-0086-3
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发表时间：
2007-01-01
影响因子：
3.4
通讯作者：
Madhyastha, M. S.
中科院分区：
文献类型：
--
作者：
Burton, E.;Yakandawala, N.;Madhyastha, M. S.
建立了一种荧光光谱测定法，用于定量生长在微滴板上的细菌生物膜。该方法涉及用小麦胚芽凝集素- alexa Fluor 488偶联物对革兰氏阴性和革兰氏阳性细菌形成的生物膜进行染色，该偶联物选择性地结合生物膜中的n-乙酰氨基葡萄糖残基。用荧光板阅读器测定染色生物膜的荧光。该方法与广泛使用的微孔板比色法进行了比较，该比色法涉及对革兰氏阴性和革兰氏阳性细菌形成的生物膜进行结晶紫染色。两种方法之间存在较强的线性相关（r(2)=0.99/0.94）。与比色法相比，荧光光谱法具有更高的灵敏度和特异性，为细菌生物膜的定量和表征提供了更好的选择。
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.