Biofilm localization in the vertical wall of shaking 96-well plates.
Biofilm localization in the vertical wall of shaking 96-well plates.
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DOI：
10.1155/2014/231083
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发表时间：
2014
期刊：
影响因子：
3.2
通讯作者：
Mergulhão FJ
中科院分区：
文献类型：
--
作者：
Gomes LC;Moreira JM;Simões M;Melo LF;Mergulhão FJ
具有96个威尔斯孔的微量滴定板由于其高通量、低成本、易于操作以及易于应用几种分析方法来评估不同的生物膜参数而越来越多地用于生物膜研究。这些方法提供了关于在每个孔中形成的生物膜的大量信息，但缺乏细节，即关于生物膜的空间位置。该位置可以通过使用光学和电子显微镜的显微镜观察来获得，但是这些技术具有较低的吞吐量和较高的成本，并且受到设备可用性的影响。这项工作描述了一种差分结晶紫（CV）染色方法，使大肠杆菌生物膜的空间位置的确定形成在振荡96孔板的垂直壁。结果表明，生物膜在壁面上分布不均匀，在气液界面附近细胞聚集较密集。扫描电子显微镜和相关性的结果得到证实，被发现之间的生物膜的积累和壁剪切应变率由计算流体动力学。开发的方法是更快，更便宜，并具有更高的吞吐量比现有的方法可用于空间定位的生物膜在微量滴定板。
Microtiter plates with 96 wells are being increasingly used for biofilm studies due to their high throughput, low cost, easy handling, and easy application of several analytical methods to evaluate different biofilm parameters. These methods provide bulk information about the biofilm formed in each well but lack in detail, namely, regarding the spatial location of the biofilms. This location can be obtained by microscopy observation using optical and electron microscopes, but these techniques have lower throughput and higher cost and are subjected to equipment availability. This work describes a differential crystal violet (CV) staining method that enabled the determination of the spatial location of Escherichia coli biofilms formed in the vertical wall of shaking 96-well plates. It was shown that the biofilms were unevenly distributed on the wall with denser cell accumulation near the air-liquid interface. The results were corroborated by scanning electron microscopy and a correlation was found between biofilm accumulation and the wall shear strain rates determined by computational fluid dynamics. The developed method is quicker and less expensive and has a higher throughput than the existing methods available for spatial location of biofilms in microtiter plates.