Correlative Time-Resolved Fluorescence Microscopy To Assess Antibiotic Diffusion-Reaction in Biofilms
Correlative Time-Resolved Fluorescence Microscopy To Assess Antibiotic Diffusion-Reaction in Biofilms
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DOI：
10.1128/aac.00216-12
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发表时间：
2012-06-01
影响因子：
4.9
通讯作者：
Steenkeste, K.
中科院分区：
文献类型：
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作者：
Oubekka, S. Daddi;Briandet, R.;Steenkeste, K.
抗生素无法灭活生物膜中组织的体内病原体已被证明会引发慢性感染。除了涉及特定遗传或生理细胞特性的机制外，抗生素吸附和/或与生物膜成分的反应可能会降低抗生素的生物利用度，从而降低其效率。为了局部准确地评估抗生素扩散反应，我们首次使用了一套先进的荧光显微工具（光漂白后的荧光恢复、荧光相关光谱和荧光寿命成像），这些工具提供了常用的延时共焦成像方法所不具备的时空分辨率。这套技术用于表征两种金黄色葡萄球菌人类分离株形成的生物膜中荧光标记的万古霉素的动态。我们证明，在万古霉素的治疗浓度下，生物膜基质并不是抗生素扩散反应的障碍，抗生素可以通过生物结构到达所有细胞。
The failure of antibiotics to inactivate in vivo pathogens organized in biofilms has been shown to trigger chronic infections. In addition to mechanisms involving specific genetic or physiological cell properties, antibiotic sorption and/or reaction with biofilm components may lessen the antibiotic bioavailability and consequently decrease their efficiency. To assess locally and accurately the antibiotic diffusion-reaction, we used for the first time a set of advanced fluorescence microscopic tools (fluorescence recovery after photobleaching, fluorescence correlation spectroscopy, and fluorescence lifetime imaging) that offer a spatiotemporal resolution not available with the commonly used time-lapse confocal imaging method. This set of techniques was used to characterize the dynamics of fluorescently labeled vancomycin in biofilms formed by two Staphylococcus aureus human isolates. We demonstrate that, at therapeutic concentrations of vancomycin, the biofilm matrix was not an obstacle to the diffusion-reaction of the antibiotic that can reach all cells through the biostructure.