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生命科学中用于可逆和不可逆电穿孔的各种治疗室设计的优点和局限性。

Advantages and limitations of various treatment chamber designs for reversible and irreversible electroporation in life sciences.

作者信息

Zand Elena, Schottroff Felix, Steinacker Elisabeth, Mae-Gano Jennifer, Schoenher Christoph, Wimberger Terje, Wassermann Klemens J, Jaeger Henry

机构信息

Institute of Food Technology, University of Natural Resources and Life Sciences (BOKU), Vienna, Austria.

Institute of Food Technology, University of Natural Resources and Life Sciences (BOKU), Vienna, Austria; BOKU Core Facility Food & Bio Processing, University of Natural Resources and Life Sciences (BOKU), Vienna, Austria.

出版信息

Bioelectrochemistry. 2021 Oct;141:107841. doi: 10.1016/j.bioelechem.2021.107841. Epub 2021 May 11.

Abstract

The fundamental mechanisms of pulsed electric fields on biological cells are not yet fully elucidated, though it is apparent that membrane electroporation plays a crucial role. Little is known about treatment-chamber-specific effects, and systematic studies are scarce. Thus, the present study evaluates the (dis-)advantages of various treatment chamber designs for liquid applications at differing scales. Three chambers, namely parallel plate microfluidic (V̇: 0.1 ml/min; titanium electrodes), co-linear meso (V̇: 5.0 ml/min; stainless steel electrodes), and co-linear macro (V̇: 83.3 ml/min; stainless steel electrodes) chambers, were studied. Electroporation effects on Escherichia coli in media with 0.1-10.0 mS/cm were evaluated by plate counts and flow cytometry at 8, 16, and 20 kV/cm. For the microfluidic chamber, predominantly irreversible electroporation (2.5 logs reductions) was seen at 0.1 mS/cm, while high irreversible electroporation (4.2 logs reductions) at 10.0 mS/cm was observed for the macro chamber. The meso chamber indicated a similar trend towards increased conductivity, even though only low inactivation levels were present. Variation in conductivity and electrode configuration or area likely induces effects resulting in distinct electroporation levels, as observed for the micro and macro chamber. Suitable application scenarios, depending on targeted electroporation effects, were suggested.

摘要

尽管很明显膜电穿孔起着关键作用,但脉冲电场作用于生物细胞的基本机制尚未完全阐明。关于处理腔室特定的影响知之甚少,且缺乏系统性研究。因此,本研究评估了不同规模下用于液体应用的各种处理腔室设计的(优)缺点。研究了三个腔室,即平行板微流控腔室(流速:0.1毫升/分钟;钛电极)、共线中尺度腔室(流速:5.0毫升/分钟;不锈钢电极)和共线大尺度腔室(流速:83.3毫升/分钟;不锈钢电极)。通过平板计数和流式细胞术,在8、16和20千伏/厘米的电场强度下,评估了0.1 - 10.0毫西门子/厘米介质中脉冲电场对大肠杆菌的电穿孔效应。对于微流控腔室,在0.1毫西门子/厘米时主要观察到不可逆电穿孔(菌落数减少2.5个对数级),而对于大尺度腔室,在10.0毫西门子/厘米时观察到高不可逆电穿孔(菌落数减少4.2个对数级)。中尺度腔室显示出随着电导率增加的类似趋势,尽管只有低失活水平。如在微尺度和大尺度腔室中所观察到的,电导率以及电极配置或面积的变化可能会引发导致不同电穿孔水平的效应。根据目标电穿孔效应,提出了合适的应用场景。

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