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High-density distributed electrode network, a multi-functional electroporation method for delivery of molecules of different sizes.

Wu M, Zhao D, Zhong W, Yan H, Wang X, Liang Z, Li Z - Sci Rep (2013)

Bottom Line: We present a multi-functional electroporation method for delivery of biomolecule utilizing a high-density distributed electrode network (HDEN) under tri-phase electric stimulation.The HDEN device, with which drastic pH change during the electroporation was avoided,was demonstrated to be highly effective for transfection of not only DNA plasmids and small interfering RNAs (siRNA), but also a small molecular anti-cancer drug, into cells in adjustable volumes of cell suspension.The method constitutes a very flexible electroporation approach in a wide range of in vitro or ex vivo scenarios in various tubes, standard multi-well plates as well as flow chambers.

View Article: PubMed Central - PubMed

Affiliation: 1] Institute of Microelectronics, National Key Laboratory of Science and Technology on Micro/Nano Fabrication, Peking University, Beijing 100871, China [2].

ABSTRACT
We present a multi-functional electroporation method for delivery of biomolecule utilizing a high-density distributed electrode network (HDEN) under tri-phase electric stimulation. The HDEN device, with which drastic pH change during the electroporation was avoided,was demonstrated to be highly effective for transfection of not only DNA plasmids and small interfering RNAs (siRNA), but also a small molecular anti-cancer drug, into cells in adjustable volumes of cell suspension. The method constitutes a very flexible electroporation approach in a wide range of in vitro or ex vivo scenarios in various tubes, standard multi-well plates as well as flow chambers.

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The design of HDEN electroporation devices.(A) Detailed illustration of a naked single HDEN device and the schematic view of electroporation procedure compatible with 96-well-plate. (B) The distribution of electrode network and schematic diagram of tri-phase electroporation method. Electrodes colored in blue, red and green are connected to pole I, II and III respectively, and electric polarity is alternated during three phases. (C) Simulation of electric field distribution during three phases and the equivalent field intensity. The abbreviations t1, t2, t3 represent three phases, and Max means the equivalent electric field if three phases are merged.
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f1: The design of HDEN electroporation devices.(A) Detailed illustration of a naked single HDEN device and the schematic view of electroporation procedure compatible with 96-well-plate. (B) The distribution of electrode network and schematic diagram of tri-phase electroporation method. Electrodes colored in blue, red and green are connected to pole I, II and III respectively, and electric polarity is alternated during three phases. (C) Simulation of electric field distribution during three phases and the equivalent field intensity. The abbreviations t1, t2, t3 represent three phases, and Max means the equivalent electric field if three phases are merged.

Mentions: As shown in Figure 1A, thirty-seven pillared electrodes were joined and welded on a printed circuit board (PCB). The electrodes were modified from bio-compatible stainless steel acupuncture needles. When electroporation was performed, pillared electrodes were inserted into the well of a 96-well plate or single tubes and immersed in cell suspension with various sample volumes. Pillared electrodes were arranged as a hexagonal cellular array (Figure 1B) and connected to three polarities by the patterned wires on the PCB (as shown in Supplementary Figure 1) to ensure the relatively uniform coverage of electric field. An improved tri-phase electric stimulation mode was proposed (Figure 1B). The electrodes were divided into three groups, as colored in blue, red and green, and each group was connected to anode alternately during a three phasic stimulation controlled by a circuit consisted of three switches. The circuit detected the electric stimulation, and then alternated the switches so that the polarity would have been shifted before that next pulse starts. The electric field generated by the electrode network is simulated as shown in Figure 1C. For a single pulse, electric traps emerged between homo-electric-polar electrodes where electric field intensity was approximately zero. However, positions of electric traps varied among three phases, and electric field covered most areas of the entire well by tri-phase method. Compared with single-phase electroporation mode, tri-phase stimulation generated a rather uniform electric field and the electric field covered the entire area (Supplementary Figure 2).


High-density distributed electrode network, a multi-functional electroporation method for delivery of molecules of different sizes.

Wu M, Zhao D, Zhong W, Yan H, Wang X, Liang Z, Li Z - Sci Rep (2013)

The design of HDEN electroporation devices.(A) Detailed illustration of a naked single HDEN device and the schematic view of electroporation procedure compatible with 96-well-plate. (B) The distribution of electrode network and schematic diagram of tri-phase electroporation method. Electrodes colored in blue, red and green are connected to pole I, II and III respectively, and electric polarity is alternated during three phases. (C) Simulation of electric field distribution during three phases and the equivalent field intensity. The abbreviations t1, t2, t3 represent three phases, and Max means the equivalent electric field if three phases are merged.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3842547&req=5

f1: The design of HDEN electroporation devices.(A) Detailed illustration of a naked single HDEN device and the schematic view of electroporation procedure compatible with 96-well-plate. (B) The distribution of electrode network and schematic diagram of tri-phase electroporation method. Electrodes colored in blue, red and green are connected to pole I, II and III respectively, and electric polarity is alternated during three phases. (C) Simulation of electric field distribution during three phases and the equivalent field intensity. The abbreviations t1, t2, t3 represent three phases, and Max means the equivalent electric field if three phases are merged.
Mentions: As shown in Figure 1A, thirty-seven pillared electrodes were joined and welded on a printed circuit board (PCB). The electrodes were modified from bio-compatible stainless steel acupuncture needles. When electroporation was performed, pillared electrodes were inserted into the well of a 96-well plate or single tubes and immersed in cell suspension with various sample volumes. Pillared electrodes were arranged as a hexagonal cellular array (Figure 1B) and connected to three polarities by the patterned wires on the PCB (as shown in Supplementary Figure 1) to ensure the relatively uniform coverage of electric field. An improved tri-phase electric stimulation mode was proposed (Figure 1B). The electrodes were divided into three groups, as colored in blue, red and green, and each group was connected to anode alternately during a three phasic stimulation controlled by a circuit consisted of three switches. The circuit detected the electric stimulation, and then alternated the switches so that the polarity would have been shifted before that next pulse starts. The electric field generated by the electrode network is simulated as shown in Figure 1C. For a single pulse, electric traps emerged between homo-electric-polar electrodes where electric field intensity was approximately zero. However, positions of electric traps varied among three phases, and electric field covered most areas of the entire well by tri-phase method. Compared with single-phase electroporation mode, tri-phase stimulation generated a rather uniform electric field and the electric field covered the entire area (Supplementary Figure 2).

Bottom Line: We present a multi-functional electroporation method for delivery of biomolecule utilizing a high-density distributed electrode network (HDEN) under tri-phase electric stimulation.The HDEN device, with which drastic pH change during the electroporation was avoided,was demonstrated to be highly effective for transfection of not only DNA plasmids and small interfering RNAs (siRNA), but also a small molecular anti-cancer drug, into cells in adjustable volumes of cell suspension.The method constitutes a very flexible electroporation approach in a wide range of in vitro or ex vivo scenarios in various tubes, standard multi-well plates as well as flow chambers.

View Article: PubMed Central - PubMed

Affiliation: 1] Institute of Microelectronics, National Key Laboratory of Science and Technology on Micro/Nano Fabrication, Peking University, Beijing 100871, China [2].

ABSTRACT
We present a multi-functional electroporation method for delivery of biomolecule utilizing a high-density distributed electrode network (HDEN) under tri-phase electric stimulation. The HDEN device, with which drastic pH change during the electroporation was avoided,was demonstrated to be highly effective for transfection of not only DNA plasmids and small interfering RNAs (siRNA), but also a small molecular anti-cancer drug, into cells in adjustable volumes of cell suspension. The method constitutes a very flexible electroporation approach in a wide range of in vitro or ex vivo scenarios in various tubes, standard multi-well plates as well as flow chambers.

Show MeSH