Cell electroporation with a three-dimensional microelectrode array on a printed circuit board

Xu, Youchun; Su, Shisheng; Zhou, Changcheng; Lü, Ying; Xing, Wanli

doi:10.1016/j.bioelechem.2014.10.002

Cited by 17 publications

(15 citation statements)

References 35 publications

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“…Independent on the application, the electroporation protocols are constantly being developed to improve the efficacy of the treatment 23 24 . The efficient permeabilization can be performed only after careful selection of the pulse parameters, such as the pulse amplitude, shape, pulse duration, repetition rate and the total number of pulses, which have major impact on the outcome of the treatment 25 26 27 .…”

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confidence: 99%

Pulsed Electromagnetic Field Assisted in vitro Electroporation: A Pilot Study

Novickij

Grainys

Lastauskienė

et al. 2016

Sci Rep

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Electroporation is a phenomenon occurring due to exposure of cells to Pulsed Electric Fields (PEF) which leads to increase of membrane permeability. Electroporation is used in medicine, biotechnology, and food processing. Recently, as an alternative to electroporation by PEF, Pulsed ElectroMagnetic Fields (PEMF) application causing similar biological effects was suggested. Since induced electric field in PEMF however is 2–3 magnitudes lower than in PEF electroporation, the membrane permeabilization mechanism remains hypothetical. We have designed pilot experiments where Saccharomyces cerevisiae and Candida lusitaniae cells were subjected to single 100–250 μs electrical pulse of 800 V with and without concomitant delivery of magnetic pulse (3, 6 and 9 T). As expected, after the PEF pulses only the number of Propidium Iodide (PI) fluorescent cells has increased, indicative of membrane permeabilization. We further show that single sub-millisecond magnetic field pulse did not cause detectable poration of yeast. Concomitant exposure of cells to pulsed electric (PEF) and magnetic field (PMF) however resulted in the increased number PI fluorescent cells and reduced viability. Our results show increased membrane permeability by PEF when combined with magnetic field pulse, which can explain electroporation at considerably lower electric field strengths induced by PEMF compared to classical electroporation.

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confidence: 99%

Pulsed Electromagnetic Field Assisted in vitro Electroporation: A Pilot Study

Novickij

Grainys

Lastauskienė

et al. 2016

Sci Rep

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“…Two or more materials could be directly used in turn by 3D printing to fabricate multiple-functional lab-on-a-chip devices. More importantly, microfluidic chips fabricated by 3D printing could mimic standard micro-titer plates and seamlessly incorporate them with control systems, solving the interfacing issue in the micro-system [39][40][41].…”

Section: Introductionmentioning

confidence: 99%

Chemical and biochemical analysis on lab-on-a-chip devices fabricated using three-dimensional printing

Zhang

2016

TrAC Trends in Analytical Chemistry

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“…Later, Xu et al . developed a printed circuit board (PCB)-based suspended-drop system, in which each electroporation unit can be individually controlled 34 . Unfortunately, the throughput of these systems was difficult to increase due to the integration with conventional microtiter plates, and cells still must be in a suspended form for electroporation.…”

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confidence: 99%

“…Second, in a cell microarray, all the cells are usually cultured in a homogeneous condition, which cannot eliminate cross-contaminations among cell clusters. Third, since cells were cultured and electroporated directly on the electrodes, the changes of pH or temperature induced by electrolysis could damage cells 34 . Also, if the electrodes were fabricated using metals, the observation of cells using an inverted microscope become impossible 26 35 .…”

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confidence: 99%

High-throughput in situ cell electroporation microsystem for parallel delivery of single guide RNAs into mammalian cells

Bian

Zhou

et al. 2017

Sci Rep

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Arrayed genetic screens mediated by the CRISPR/Cas9 technology with single guide RNA (sgRNA) libraries demand a high-throughput platform capable of transfecting diverse cell types at a high efficiency in a genome-wide scale for detection and analysis of sophisticated cellular phenotypes. Here we developed a high-throughput in situ cell electroporation (HiCEP) microsystem which leveraged the superhydrophobic feature of the microwell array to achieve individually controlled conditions in each microwell and coupled an interdigital electrode array chip with the microwells in a modular-based scheme for highly efficient delivery of exogenous molecules into cells. Two plasmids encoding enhanced green and red fluorescent proteins (EGFP and ERFP), respectively, were successfully electroporated into attached HeLa cells on a 169-microwell array chip with transfection efficiencies of 71.6 ± 11.4% and 62.9 ± 2.7%, and a cell viability above 95%. We also successfully conducted selective electroporation of sgRNA into 293T cells expressing the Cas9 nuclease in a high-throughput manner and observed the four-fold increase of the GFP intensities due to the repair of the protein coding sequences mediated by the CRISPR/Cas9 system. This study proved that this HiCEP system has the great potential to be used for arrayed functional screens with genome-wide CRISPR libraries on hard-to-transfect cells in the future.

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Cell electroporation with a three-dimensional microelectrode array on a printed circuit board

Cited by 17 publications

References 35 publications

Pulsed Electromagnetic Field Assisted in vitro Electroporation: A Pilot Study

Pulsed Electromagnetic Field Assisted in vitro Electroporation: A Pilot Study

Chemical and biochemical analysis on lab-on-a-chip devices fabricated using three-dimensional printing

High-throughput in situ cell electroporation microsystem for parallel delivery of single guide RNAs into mammalian cells

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