A porous membrane-based culture substrate for localized in situ electroporation of adherent mammalian cells

Takeshi Ishibashi, Kimiyasu Takoh, Hirokazu Kaji, Takashi Abe, Matsuhiko Nishizawa

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

We report herein the invention of and proof of function for a porous membrane-based electroporation device that can deliver molecules into spatially restricted and predefined areas of a cell monolayer. The device's cell culture substrate is a microporous alumina membrane (pore size 0.02 μm), with an underlying thin poly-(dimethylsiloxane) (PDMS) film that has one or more holes with diameters in the one-tenth millimeter range. When a transient electric field is generated between the device's two planar electrodes - one of which is placed above the cells and the other below the PDMS layer - the filed condenses only in the volume defined by hole in the PDMS film and therefore localized electroporation can occur. We demonstrate that Lucifer Yellow (LY) and plasmid DNA are selectively introduced into only those HeLa cells located above the holes. Using the device containing a PDMS film with multiple holes, a patterned array of LY-stained cells was resulted. Compared with the operation of and the results obtained from a conventional cuvette electroporation device, our device greatly decreases the necessary operating voltage, can be used with cells attached to a substrate, and increases the number of conditions that can be screened in a single experiment. Finally, since a PDMS film with different sized holes, produces different localized electric field strengths, it is possible to determine the optimum electroporetic conditions in a single experiment.

Original languageEnglish
Pages (from-to)5-11
Number of pages7
JournalSensors and Actuators, B: Chemical
Volume128
Issue number1
DOIs
Publication statusPublished - 2007 Dec 12

Keywords

  • Cell culture
  • Electroporation
  • PDMS

ASJC Scopus subject areas

  • Electronic, Optical and Magnetic Materials
  • Instrumentation
  • Condensed Matter Physics
  • Surfaces, Coatings and Films
  • Metals and Alloys
  • Electrical and Electronic Engineering
  • Materials Chemistry

Fingerprint Dive into the research topics of 'A porous membrane-based culture substrate for localized in situ electroporation of adherent mammalian cells'. Together they form a unique fingerprint.

  • Cite this