Ogawa M, Tani K, Yamaguchi N, Nasu M
Environmental Science and Microbiology, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan.
J Appl Microbiol. 2003;95(1):120-8. doi: 10.1046/j.1365-2672.2003.01950.x.
To develop a rapid and simple method for the assessment of metabolic activity of bacteria in natural environment.
A rapid and simple multicolour digital image analysis system for enumerating viable bacteria based on active fluorescent staining has been developed. This system can accurately differentiate actively respiring bacteria and non-respiring bacteria by distinctive colour information in the digital image captured by an epifluorescence microscope equipped with low magnification objective lens. An algorithm to distinguish bacteria from considerable detritus, which produced bright fluorescence in different colours, by colour segmentation has also been developed. This system was applied to river water samples, and the total and respiratory active bacterial counts by digital image analysis were highly related to those by epifluorescence microscopy (r2 = 0.96 and 0.93, respectively).
This system allowed the rapid and simple differentiation of bacteria from detritus and concurrent assessment of their metabolic activity, with results being available within 1 h.
The low magnification image analysis allows more rapid and simple quantification of bacteria and could considerably decrease image data amount and acquisition time. This system could be easily applied to the rapid analysis of microbes in various environments.
开发一种快速、简便的方法来评估自然环境中细菌的代谢活性。
已开发出一种基于活性荧光染色的用于计数活细菌的快速、简便的多色数字图像分析系统。该系统可通过配备低倍物镜的落射荧光显微镜拍摄的数字图像中的独特颜色信息，准确区分活跃呼吸的细菌和不呼吸的细菌。还开发了一种通过颜色分割从大量产生不同颜色明亮荧光的碎屑中区分细菌的算法。该系统应用于河水样本，通过数字图像分析得到的总细菌数和呼吸活性细菌数与落射荧光显微镜得到的结果高度相关（分别为r2 = 0.96和0.93）。
该系统能够快速、简便地将细菌与碎屑区分开来，并同时评估它们的代谢活性，1小时内即可获得结果。
低倍图像分析能够更快速、简便地对细菌进行定量，并且可以大幅减少图像数据量和采集时间。该系统可轻松应用于各种环境中微生物的快速分析。
