Flow-type electroporation chips for gene transfection

Yu-Cheng Lin, Chung Min Jen, Ming Yuan Huang, Xi-Zhang Lin

Research output: Contribution to journalConference article

1 Citation (Scopus)

Abstract

Electroporation is a technique with which DNA molecules can be delivered into cells in a chamber using high electric field pulses. The limited amount of target cells and the potential risk from the high voltage are the two drawbacks in this technique. This study aimed to fabricate an electroporation chip to manage large amount of cells continuously with a lower applied voltage. The electroporation chip, consisting of a micro-channel with thin film electrodes made of gold or platinum wire electrodes on both sides, was fabricated on PMMA material using evaporation, photolithography, wet-etching, lift-off, and fusion-bonding methods. The suspension fluid of Huh-7 cell lines (1×106 cells/ml) mixed with 10 μg plasmids equipped with lacZ genes in a volume of 500 μl flowed through the channel with a variety of flow rates under a series of square pulses with 1-10 ms in width and 10 Hz in frequency. Trypan-blue staining assessed the viability of the cells treated with pulses. The transfection rate was evaluated with blue-staining cells under X-Gal stain 24 hours later. The dimensions of the channel were 5 mm wide, 0.2 mm high, and 25 mm long. Two types of electrodes, parallel-plate type and parallel-line type electrodes, were fabricated and tested in these experiments. The fabricated microchip can deliver genes into the flowing cells. The electric pulse frequency that determines the shock number for each cell for a fixed flow rate can be optimized for better transfection and survival rates.

Original languageEnglish
Pages (from-to)194-198
Number of pages5
JournalProceedings of SPIE - The International Society for Optical Engineering
Volume4177
Publication statusPublished - 2000 Dec 1
EventMicrofluidic Devices and Systems III - Santa Clara, CA, USA
Duration: 2000 Sep 182000 Sep 19

Fingerprint

genes
Chip
Genes
chips
Gene
Electrodes
Cell
cells
Electrode
Cells
Flow rate
Trypan Blue
electrodes
Wet etching
Electric potential
Photolithography
Polymethyl Methacrylate
staining
Platinum
Gold

All Science Journal Classification (ASJC) codes

  • Electronic, Optical and Magnetic Materials
  • Condensed Matter Physics
  • Computer Science Applications
  • Applied Mathematics
  • Electrical and Electronic Engineering

Cite this

@article{cf8a5c29ff0e4c0f9eae7d0fa9d9bfb9,
title = "Flow-type electroporation chips for gene transfection",
abstract = "Electroporation is a technique with which DNA molecules can be delivered into cells in a chamber using high electric field pulses. The limited amount of target cells and the potential risk from the high voltage are the two drawbacks in this technique. This study aimed to fabricate an electroporation chip to manage large amount of cells continuously with a lower applied voltage. The electroporation chip, consisting of a micro-channel with thin film electrodes made of gold or platinum wire electrodes on both sides, was fabricated on PMMA material using evaporation, photolithography, wet-etching, lift-off, and fusion-bonding methods. The suspension fluid of Huh-7 cell lines (1×106 cells/ml) mixed with 10 μg plasmids equipped with lacZ genes in a volume of 500 μl flowed through the channel with a variety of flow rates under a series of square pulses with 1-10 ms in width and 10 Hz in frequency. Trypan-blue staining assessed the viability of the cells treated with pulses. The transfection rate was evaluated with blue-staining cells under X-Gal stain 24 hours later. The dimensions of the channel were 5 mm wide, 0.2 mm high, and 25 mm long. Two types of electrodes, parallel-plate type and parallel-line type electrodes, were fabricated and tested in these experiments. The fabricated microchip can deliver genes into the flowing cells. The electric pulse frequency that determines the shock number for each cell for a fixed flow rate can be optimized for better transfection and survival rates.",
author = "Yu-Cheng Lin and Jen, {Chung Min} and Huang, {Ming Yuan} and Xi-Zhang Lin",
year = "2000",
month = "12",
day = "1",
language = "English",
volume = "4177",
pages = "194--198",
journal = "Proceedings of SPIE - The International Society for Optical Engineering",
issn = "0277-786X",
publisher = "SPIE",

}

Flow-type electroporation chips for gene transfection. / Lin, Yu-Cheng; Jen, Chung Min; Huang, Ming Yuan; Lin, Xi-Zhang.

In: Proceedings of SPIE - The International Society for Optical Engineering, Vol. 4177, 01.12.2000, p. 194-198.

Research output: Contribution to journalConference article

TY - JOUR

T1 - Flow-type electroporation chips for gene transfection

AU - Lin, Yu-Cheng

AU - Jen, Chung Min

AU - Huang, Ming Yuan

AU - Lin, Xi-Zhang

PY - 2000/12/1

Y1 - 2000/12/1

N2 - Electroporation is a technique with which DNA molecules can be delivered into cells in a chamber using high electric field pulses. The limited amount of target cells and the potential risk from the high voltage are the two drawbacks in this technique. This study aimed to fabricate an electroporation chip to manage large amount of cells continuously with a lower applied voltage. The electroporation chip, consisting of a micro-channel with thin film electrodes made of gold or platinum wire electrodes on both sides, was fabricated on PMMA material using evaporation, photolithography, wet-etching, lift-off, and fusion-bonding methods. The suspension fluid of Huh-7 cell lines (1×106 cells/ml) mixed with 10 μg plasmids equipped with lacZ genes in a volume of 500 μl flowed through the channel with a variety of flow rates under a series of square pulses with 1-10 ms in width and 10 Hz in frequency. Trypan-blue staining assessed the viability of the cells treated with pulses. The transfection rate was evaluated with blue-staining cells under X-Gal stain 24 hours later. The dimensions of the channel were 5 mm wide, 0.2 mm high, and 25 mm long. Two types of electrodes, parallel-plate type and parallel-line type electrodes, were fabricated and tested in these experiments. The fabricated microchip can deliver genes into the flowing cells. The electric pulse frequency that determines the shock number for each cell for a fixed flow rate can be optimized for better transfection and survival rates.

AB - Electroporation is a technique with which DNA molecules can be delivered into cells in a chamber using high electric field pulses. The limited amount of target cells and the potential risk from the high voltage are the two drawbacks in this technique. This study aimed to fabricate an electroporation chip to manage large amount of cells continuously with a lower applied voltage. The electroporation chip, consisting of a micro-channel with thin film electrodes made of gold or platinum wire electrodes on both sides, was fabricated on PMMA material using evaporation, photolithography, wet-etching, lift-off, and fusion-bonding methods. The suspension fluid of Huh-7 cell lines (1×106 cells/ml) mixed with 10 μg plasmids equipped with lacZ genes in a volume of 500 μl flowed through the channel with a variety of flow rates under a series of square pulses with 1-10 ms in width and 10 Hz in frequency. Trypan-blue staining assessed the viability of the cells treated with pulses. The transfection rate was evaluated with blue-staining cells under X-Gal stain 24 hours later. The dimensions of the channel were 5 mm wide, 0.2 mm high, and 25 mm long. Two types of electrodes, parallel-plate type and parallel-line type electrodes, were fabricated and tested in these experiments. The fabricated microchip can deliver genes into the flowing cells. The electric pulse frequency that determines the shock number for each cell for a fixed flow rate can be optimized for better transfection and survival rates.

UR - http://www.scopus.com/inward/record.url?scp=0034547215&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034547215&partnerID=8YFLogxK

M3 - Conference article

AN - SCOPUS:0034547215

VL - 4177

SP - 194

EP - 198

JO - Proceedings of SPIE - The International Society for Optical Engineering

JF - Proceedings of SPIE - The International Society for Optical Engineering

SN - 0277-786X

ER -