Multicolour digital image analysis system for identification of bacteria and concurrent assessment of their respiratory activity |
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Authors: | Ogawa M Tani K Ochiai A Yamaguchi N Nasu M |
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Institution: | Environmental Science and Microbiology, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan. |
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Abstract: | AIMS: To develop a rapid and simple multicolour digital image analysis system for simultaneous identification of bacteria and assessment of their metabolic activity. METHODS AND RESULTS: We developed an image analyser capable of distinguishing triple-stained bacterial cells. Bacteria were stained with a nucleic acid stain, a fluorescent antibody and a fluorescent metabolic indicator for enumeration, species identification and assessment of metabolic activity. This multicolour image analyser was used to simultaneously identify Escherichia coli O157:H7 in milk samples and assess their respiratory activity. The images of the triple-stained bacteria were captured using a combination of blue light and u.v. excitation and an epifluorescence microscope and were processed by our image analyser. We found a good correlation between the counts of actively respiring (r = 0.93) and total (r = 0.94) E. coli O157:H7 measured by digital image analysis and visual observation. CONCLUSION: The multicolour digital image analysis system described here was able to quantify active pathogenic micro-organisms within 2 h. SIGNIFICANCE AND IMPACT OF THE STUDY: This multicolour image analysis allows the rapid and simultaneous quantification of bacteria, identification of species and assessment of metabolic activity. |
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Keywords: | active fluorescent staining epifluorescence microscope Escherichia coli O157:H7 fluorescent antibody staining multicolour digital image analysis rapid enumeration triple staining |
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