Quantitative and real time detection of pulsed electric field induced damage on Escherichia coli cells and sublethally injured microbial cells using flow cytometry in combination with fluorescent techniques
Quantitative and real time detection of pulsed electric field induced damage on Escherichia coli cells and sublethally injured microbial cells using flow cytometry in combination with fluorescent techniques
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DOI：
10.1016/j.foodcont.2010.10.006
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发表时间：
2011-03
期刊：
影响因子：
6
通讯作者：
Wei Zhao;Ruijin Yang;Howard Q. Zhang;Wenbin Zhang;Xiao Hua;Yali Tang
中科院分区：
文献类型：
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作者：
Wei Zhao;Ruijin Yang;Howard Q. Zhang;Wenbin Zhang;Xiao Hua;Yali Tang
以往大多数研究脉冲电场（PEF）对微生物细胞非热作用的研究方法只能揭示PEF诱导的微生物细胞明显的形态变化，或者只能区分可培养细胞和不可培养细胞。它们完全无法实时检测 PEF 应激下亚致死损伤的细胞。本研究采用流式细胞术（FCM）结合荧光技术，定量实时检测PEF对大肠杆菌细胞和亚致死损伤微生物细胞造成的损伤。目前的工作证实了电透化不是一个全有或全无事件的论点。双染细胞的 FCM 分选为 PEF 应激下微生物细胞发生亚致死损伤提供了直接证据。这项工作对PEF治疗中亚致死损伤的微生物细胞进行了定量和实时分析，并进一步揭示了中热和PEF协同灭活微生物的潜在机制。
The methods employed in most previous studies to investigate the non-thermal action of pulsed electric field (PEF) on microbial cells are only capable of revealing the obvious morphological changes of microbial cells induced by PEF, or only differentiate between culturable and not culturable cells. They are totally incapable of real time detection of sublethally injured cells under stress of PEF. In this study, flow cytometry (FCM) in combination with fluorescent techniques was used for quantitative and real time detection of PEF induced damage on Escherichia coli cells and sublethally injured microbial cells. The present work confirmed the thesis that electropermeabilisation was not an all-or-nothing-event. FCM sorting of double stained cells put forward direct evidence for the occurrence of sublethally injured microbial cells under stress of PEF. This work provided quantitative and real time analysis of sublethally injured microbial cells in PEF treatment, and further revealed the underlying mechanism on synergistic effect of moderate heat and PEF on microbial inactivation.