Microfluidic Irreversible Electroporation-A Versatile Tool to Extract Intracellular Contents of Bacteria and Yeast

被引：13
作者：
Rockenbach, Alexander [1 ]
Sudarsan, Suresh [2 ,3 ]
Berens, Judith [1 ]
Kosubek, Michael [1 ]
Lazar, Jaroslav [1 ]
Demling, Philipp [2 ]
Hanke, Rene [2 ]
Mennicken, Philip [2 ]
Ebert, Birgitta E. [2 ,4 ]
Blank, Lars M. [2 ,5 ]
Schnakenberg, Uwe [1 ]
机构：
[1] Rhein Westfal TH Aachen, Inst Mat Elect Engn 1, D-52074 Aachen, Germany
[2] Rhein Westfal TH Aachen, ABBt Aachen Biol & Biotechnol Dept, iAMB Inst Appl Microbiol, D-52074 Aachen, Germany
[3] Tech Univ Denmark, Novo Nordisk Fdn, Ctr Biosustainabil, DK-2800 Lyngby, Denmark
[4] Univ Queensland, Australian Inst Bioengn & Nanotechnol, Brisbane, Qld 4072, Australia
[5] Forschungszentrum Julich, Bioecon Sci Ctr BioSC, D-52425 Julich, Germany
来源：
METABOLITES | 2019年 / 9卷 / 10期
关键词：
irreversible electroporation; microfluidics; microelectrodes; pulsed electric field electroporation; intracellular metabolites; enzymes; quenching; E; coli; S; cerevisiae; PULSED ELECTRIC-FIELDS; CELL-LYSIS; SACCHAROMYCES-CEREVISIAE; ESCHERICHIA-COLI; INACTIVATION; CHIP; METABOLOMICS; MICROORGANISMS; DEVICE; GENES;
D O I：
10.3390/metabo9100211
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
Exploring the dynamic behavior of cellular metabolism requires a standard laboratory method that guarantees rapid sampling and extraction of the cellular content. We propose a versatile sampling technique applicable to cells with different cell wall and cell membrane properties. The technique is based on irreversible electroporation with simultaneous quenching and extraction by using a microfluidic device. By application of electric pulses in the millisecond range, permanent lethal pores are formed in the cell membrane of Escherichia coli and Saccharomyces cerevisiae, facilitating the release of the cellular contents; here demonstrated by the measurement of glucose-6-phosphate and the activity of the enzyme glucose-6-phosphate dehydrogenase. The successful application of this device was demonstrated by pulsed electric field treatment in a flow-through configuration of the microfluidic chip in combination with sampling, inactivation, and extraction of the intracellular content in a few seconds. Minimum electric field strengths of 10 kV/cm for E. coli and 7.5 kV/cm for yeast S. cerevisiae were required for successful cell lysis. The results are discussed in the context of applications in industrial biotechnology, where metabolomics analyses are important.
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页数：13
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