Use of Ratiometric Probes with a Spectrofluorometer for Bacterial Viability Measurement
Use of Ratiometric Probes with a Spectrofluorometer for Bacterial Viability Measurement
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DOI：
10.4014/jmb.1804.04048
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发表时间：
2018-11-01
影响因子：
2.8
通讯作者：
Lencel, Philippe
中科院分区：
文献类型：
--
作者：
Cleach, Jerome;Watier, Denis;Lencel, Philippe
微生物存活力的评估在许多工业领域中是有用的。已经开发了大量与使用荧光探针相关的方法，包括荧光测定法、荧光显微镜和细胞计数法。本研究采用微体积荧光分光光度计测定大肠杆菌的膜电位变化。为了评价该装置的灵敏度，测定了E.使用离子载体试剂：羰腈3-氯苯腙人工破坏大肠杆菌。用两种比率法测定膜电位：罗丹明123/4 '，6-二脒基-2-苯基吲哚组合和JC-10比率探针。利用这些方法研究了冷冻对大肠杆菌的影响。大肠杆菌，并与常规计数法进行了比较。结果表明，这是有益的，使用这种紧凑，易于使用，和廉价的荧光分光光度计，以评估细菌细胞的活力通过其膜电位。
Assessment of microorganism viability is useful in many industrial fields. A large number of methods associated with the use of fluorescent probes have been developed, including fluorimetry, fluorescence microscopy, and cytometry. In this study, a microvolume spectrofluorometer was used to measure the membrane potential variations of Escherichia coli. In order to estimate the sensitivity of the device, the membrane potential of E. coli was artificially disrupted using an ionophore agent: carbonyl cyanide 3-chlorophenylhydrazone. The membrane potential was evaluated using two ratiometric methods: a Rhodamine 123/4',6-diamidino-2-phenylindole combination and a JC-10 ratiometric probe. These methods were used to study the impact of freezing on E. coli, and were compared with the conventional enumeration method. The results showed that it was beneficial to use this compact, easy-to-use, and inexpensive spectrofluorometer to assess the viability of bacterial cells via their membrane potential.