Department of Environmental Microbiology, UFZ - Helmholtz-Centre for Environmental Research Leipzig, Germany.
Department of Environmental Microbiology, UFZ - Helmholtz-Centre for Environmental Research Leipzig, Germany ; Institute of Environmental and Sustainable Chemistry, TU Braunschweig Braunschweig, Germany.
Front Microbiol. 2014 Jun 4;5:273. doi: 10.3389/fmicb.2014.00273. eCollection 2014.
Optical characteristics of individual bacterial cells of natural communities can be measured with flow cytometry (FCM) in high throughput. The resulting data are visualized in cytometric histograms. These histograms represent individual cytometric fingerprints of microbial communities, e.g., at certain time points or microenvironmental conditions. Up to now four tools for analyzing the variation in these cytometric fingerprints are available but have not yet been systematically compared regarding application: Dalmatian Plot, Cytometric Histogram Image Comparison (CHIC), Cytometric Barcoding (CyBar), and FlowFP. In this article these tools were evaluated concerning (i) the required experience of the operator in handling cytometric data sets, (ii) the detection level of changes, (iii) time demand for analysis, and (iv) software requirements. As an illustrative example, FCM was used to characterize the microbial community structure of electroactive microbial biofilms. Their cytometric fingerprints were determined, analyzed with all four tools, and correlated to experimental and functional parameters. The source of inoculum (four different types of wastewater samples) showed the strongest influence on the microbial community structure and biofilm performance while the choice of substrate (acetate or lactate) had no significant effect in the present study. All four evaluation tools were found suitable to monitor structural changes of natural microbial communities. The Dalmatian Plot was shown to be most sensitive to operator impact but nevertheless provided an overview on community shifts. CHIC, CyBar, and FlowFP showed less operator dependence and gave highly resolved information on community structure variation on different detection levels. In conclusion, experimental and productivity parameters correlated with the biofilm structures and practical process integration details were available from cytometric fingerprint analysis.
利用流式细胞术（FCM）可以高通量地测量自然群落中单个细菌细胞的光学特性。所得数据以细胞计量直方图的形式可视化。这些直方图代表微生物群落的个体细胞计量指纹，例如，在特定时间点或微环境条件下。目前有四种用于分析这些细胞计量指纹变化的工具，但尚未就应用进行系统比较：Dalmatian Plot、Cytometric Histogram Image Comparison (CHIC)、Cytometric Barcoding (CyBar) 和 FlowFP。在本文中，这些工具在以下几个方面进行了评估：（i）操作员在处理细胞计量数据集方面所需的经验；（ii）检测变化的水平；（iii）分析所需的时间；（iv）软件要求。作为一个说明性的例子，使用 FCM 来描述电活性微生物生物膜的微生物群落结构。确定它们的细胞计量指纹，用所有四种工具进行分析，并与实验和功能参数相关联。接种物的来源（四种不同类型的废水样本）对微生物群落结构和生物膜性能的影响最强，而在本研究中，底物（乙酸盐或乳酸盐）的选择没有显著影响。所有四种评估工具都被发现适合监测自然微生物群落的结构变化。Dalmatian Plot 被证明对操作员的影响最敏感，但仍提供了社区变化的概述。CHIC、CyBar 和 FlowFP 显示出较少的操作员依赖性，并提供了关于不同检测水平下群落结构变化的高度解析信息。总之，从细胞计量指纹分析中可以获得与生物膜结构相关的实验和生产力参数以及实际过程集成细节。
