Development and laboratory-scale testing of a fully automated online flow cytometer for drinking water analysis
Development and laboratory-scale testing of a fully automated online flow cytometer for drinking water analysis
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DOI：
10.1002/cyto.a.22048
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发表时间：
2012-06-01
期刊：
影响因子：
3.7
通讯作者：
Sonnleitner, Bernhard
中科院分区：
文献类型：
--
作者：
Hammes, Frederik;Broger, Tobias;Sonnleitner, Bernhard
准确和灵敏的在线检测工具将有利于水生微生物学的基础研究和实际应用。在这里，我们描述了在线流式细胞仪（FCM）的开发和测试，在饮用水微生物学领域的特定用途。该系统采用全自动采样和荧光标记的细菌核酸与分析在5分钟的时间间隔超过24小时。实验室规模的测试表明，在广泛的浓度范围（1 × 1031 × 106个细胞mL-1）内对细菌进行灵敏的检测（误差< 5%），特别是能够跟踪水样中的逐渐变化和戏剧性事件。该系统进行了测试，细菌纯培养物以及土著微生物群落从天然水样。此外，我们证明了使用单一荧光染料（例如，SYBR绿色I）或两种染料（SYBR绿色I和碘化丙啶）的组合，从而拓宽了该系统的应用可能性。本文所述的在线FCM方法具有相当大的潜力，用于常规和连续监测饮用水，优化特定的饮用水过程，如生物过滤或消毒，以及一般的水生微生物学研究。(c)2012年国际细胞计数促进学会
Accurate and sensitive online detection tools would benefit both fundamental research and practical applications in aquatic microbiology. Here, we describe the development and testing of an online flow cytometer (FCM), with a specific use foreseen in the field of drinking water microbiology. The system incorporated fully automated sampling and fluorescent labeling of bacterial nucleic acids with analysis at 5-min intervals for periods in excess of 24 h. The laboratory scale testing showed sensitive detection (< 5% error) of bacteria over a broad concentration range (1 X 1031 X 106 cells mL-1) and particularly the ability to track both gradual changes and dramatic events in water samples. The system was tested with bacterial pure cultures as well as indigenous microbial communities from natural water samples. Moreover, we demonstrated the possibility of using either a single fluorescent dye (e.g., SYBR Green I) or a combination of two dyes (SYBR Green I and Propidium Iodide), thus broadening the application possibilities of the system. The online FCM approach described herein has considerable potential for routine and continuous monitoring of drinking water, optimization of specific drinking water processes such as biofiltration or disinfection, as well as aquatic microbiology research in general. (c) 2012 International Society for Advancement of Cytometry