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利用流式细胞术对水生生物膜进行表征

Characterization of Aquatic Biofilms with Flow Cytometry.

作者信息

Sgier Linn, Merbt Stephanie N, Tlili Ahmed, Kroll Alexandra, Zupanic Anze

机构信息

Department of Environmental Toxicology, Eawag - Swiss Federal Institute for Aquatic Science and Technology.

Department of Environmental Toxicology, Eawag - Swiss Federal Institute for Aquatic Science and Technology;

出版信息

J Vis Exp. 2018 Jun 6(136):57655. doi: 10.3791/57655.

Abstract

Biofilms are dynamic consortia of microorganism that play a key role in freshwater ecosystems. By changing their community structure, biofilms respond quickly to environmental changes and can be thus used as indicators of water quality. Currently, biofilm assessment is mostly based on integrative and functional endpoints, such as photosynthetic or respiratory activity, which do not provide information on the biofilm community structure. Flow cytometry and computational visualization offer an alternative, sensitive, and easy-to-use method for assessment of the community composition, particularly of the photoautotrophic part of freshwater biofilms. It requires only basic sample preparation, after which the entire sample is run through the flow cytometer. The single-cell optical and fluorescent information is used for computational visualization and biological interpretation. Its main advantages over other methods are the speed of analysis and the high-information-content nature. Flow cytometry provides information on several cellular and biofilm traits in a single measurement: particle size, density, pigment content, abiotic content in the biofilm, and coarse taxonomic information. However, it does not provide information on biofilm composition on the species level. We see high potential in the use of the method for environmental monitoring of aquatic ecosystems and as an initial biofilm evaluation step that informs downstream detailed investigations by complementary and more detailed methods.

摘要

生物膜是微生物的动态聚集体,在淡水生态系统中发挥着关键作用。通过改变其群落结构,生物膜能迅速对环境变化做出反应,因此可作为水质指标。目前,生物膜评估主要基于综合和功能终点,如光合或呼吸活性,这些并不能提供生物膜群落结构的信息。流式细胞术和计算可视化提供了一种替代的、灵敏且易于使用的方法,用于评估群落组成,特别是淡水生物膜中的光合自养部分。它只需要进行基本的样品制备,之后将整个样品通过流式细胞仪。单细胞光学和荧光信息用于计算可视化和生物学解释。与其他方法相比,其主要优点是分析速度快且信息含量高。流式细胞术在一次测量中就能提供有关几种细胞和生物膜特征的信息:颗粒大小、密度、色素含量、生物膜中的非生物成分以及粗略的分类信息。然而,它无法提供物种水平上的生物膜组成信息。我们认为该方法在水生生态系统环境监测以及作为生物膜初步评估步骤方面具有很大潜力,可为后续通过补充性更强、更详细的方法进行的详细调查提供信息。

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