Flow cytometry to assess biochemical pathways in heat-stressed Cronobacter spp. (formerly Enterobacter sakazakii)
Flow cytometry to assess biochemical pathways in heat-stressed Cronobacter spp. (formerly Enterobacter sakazakii)
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DOI：
10.1111/j.1365-2672.2011.05075.x
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发表时间：
2011-09-01
影响因子：
4
通讯作者：
Jordan, K.
中科院分区：
文献类型：
--
作者：
Arku, B.;Fanning, S.;Jordan, K.
目的：使用流式细胞术（FC）为基础的方法与四个选定的荧光探针相结合，生化途径激活后的适应克罗诺菌属。致死热应激的影响。这种方法评估的生理变化，在四个菌株的克罗诺菌属。方法和结果：使用市售的活/死的活力评估荧光探针，活，受伤或死亡的细菌细胞进行了研究。使用染料标记的探针3，3 '-二己基羰花青碘化物评价细胞呼吸和膜电位，使用荧光素二乙酸酯（FDA）探针评价代谢活性，使用羧基荧光素二乙酸琥珀酰亚胺酯探针测量细胞内pH变化，并使用氢乙啶荧光探针测量活性氧。适应致死性热应激诱导的生理变化，可能会提高生存的克罗诺斯spp.结论：这些数据表明，在原位评估的生理行为的致死性应激细胞使用多参数FC是一个有用的，快速和敏感的工具，研究和评估的活力和克罗诺斯细胞的生理状态。这项研究表明，FC是一个有价值的工具，在生理方面的研究增加生存，因为亚致死适应热。
Aims: Using a flow cytometry (FC)-based approach in combination with four selected fluorescent probes, the biochemical pathway activated following the adaptation of Cronobacter spp. to lethal heat stress was investigated. This approach assessed the physiological changes induced in four strains of Cronobacter spp.Methods and Results: Using the commercially available live/dead viability assessment fluorescence probes, live, injured or dead bacterial cells were studied. Cellular respiration and membrane potential were evaluated using the dye-labelled probe 3,3'-dihexylocarbocyanine iodide, metabolic activity was evaluated using a fluorescein diacetate (FDA) probe, intracellular pH changes were measured using a carboxy-fluorescein diacetate succinimidyl ester probe, and reactive oxygen species were measured using a hydroethidine fluorescent probe. Adaptation to lethal heat stress induced physiological changes that potentially improve the survival of Cronobacter spp.Conclusions: These data showed that in situ assessment of physiological behaviour of lethally stressed cells using multiparameter FC is a useful, rapid and sensitive tool to study and assess the viability and physiological state of Cronobacter cells.Significance and Impact of the Study: This study shows that FC is a valuable tool in the study of physiological aspects of increased survival because of sublethal adaptation to heat.