Experimental Detection and Visualization of the Extracellular Matrix in Macrocolony Biofilms.
Experimental Detection and Visualization of the Extracellular Matrix in Macrocolony Biofilms.
复制标题
大菌落生物膜中细胞外基质的实验检测和可视化。
DOI：
10.1007/978-1-4939-7240-1_11
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发表时间：
2017
影响因子：
--
通讯作者：
R. Hengge
中科院分区：
文献类型：
--
作者：
D. Serra;R. Hengge
通过采用复杂的三维形态，根据它们的细胞外基质组成而变化，大菌落生物膜提供了一个独特的机会来询问特定基质成分在形成生物膜结构中的作用。在这里，我们描述了两种针对大肠杆菌优化的方法，这些方法得益于大菌落的形态学和高水平的结构组织，从而深入了解淀粉样蛋白卷曲和纤维素（大肠杆菌在生物膜中的两种主要生物膜基质元素）的产生和组装。第一种方法，即大菌落形态测定法，是基于curli和纤维素（单独或组合）在大肠杆菌大菌落中产生特定形态和刚果红染色模式的能力，然后可以将其用作生产这些基质成分的直接视觉读数。第二种方法包括对大菌落进行薄切片，再加上淀粉样蛋白卷曲体和纤维素的原位染色以及显微成像，可以获得大菌落内两种基质元素空间排列的精细细节。除了目前用于大肠杆菌和相关的curli和生产纤维素的肠杆菌科外，这两种方法都有可能适用于其他细菌物种。
By adopting elaborate three-dimensional morphologies that vary according to their extracellular matrix composition, macrocolony biofilms offer a unique opportunity to interrogate about the roles of specific matrix components in shaping biofilm architecture. Here, we describe two methods optimized for Escherichia coli that profit from morphology and the high level of structural organization of macrocolonies to gain insight into the production and assembly of amyloid curli and cellulose-the two major biofilm matrix elements of E. coli-in biofilms. The first method, the macrocolony morphology assay, is based on the ability of curli and cellulose-either alone or in combination-to generate specific morphological and Congo Red-staining patterns in E. coli macrocolonies, which can then be used as a direct visual readout for the production of these matrix components. The second method involves thin sectioning of macrocolonies, which along with in situ staining of amyloid curli and cellulose and microscopic imaging allows gaining fine details of the spatial arrangement of both matrix elements inside macrocolonies. Beyond their current use with E. coli and related curli and cellulose-producing Enterobacteriaceae, both the methods offer the potential to be adapted to other bacterial species.
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影响因子：
11.3
作者：
Mann EE;Wozniak DJ
通讯作者：
Wozniak DJ
影响因子：
10.5
作者：
Vlamakis, Hera;Aguilar, Claudio;Kolter, Roberto
通讯作者：
Kolter, Roberto
影响因子：
3.7
作者：
Reichhardt C;Jacobson AN;Maher MC;Uang J;McCrate OA;Eckart M;Cegelski L
通讯作者：
Cegelski L
影响因子：
11.1
作者：
Richter AM;Povolotsky TL;Wieler LH;Hengge R
通讯作者：
Hengge R