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Optically transparent polymer devices for in situ assessment of cell electroporation

Majhi, Amit Kumar and Thrivikraman, Greeshma and Basu, Bikramjit and Venkataraman, V (2015) Optically transparent polymer devices for in situ assessment of cell electroporation. In: EUROPEAN BIOPHYSICS JOURNAL WITH BIOPHYSICS LETTERS, 44 (1-2). pp. 57-67.

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Official URL: http://dx.doi.org/ 10.1007/s00249-014-1001-x

Abstract

In order to study cell electroporation in situ, polymer devices have been fabricated from poly-dimethyl siloxane with transparent indium tin oxide parallel plate electrodes in horizontal geometry. This geometry with cells located on a single focal plane at the interface of the bottom electrode allows a longer observation time in both transmitted bright-field and reflected fluorescence microscopy modes. Using propidium iodide (PI) as a marker dye, the number of electroporated cells in a typical culture volume of 10-100 mu l was quantified in situ as a function of applied voltage from 10 to 90 V in a series of 2-ms pulses across 0.5-mm electrode spacing. The electric field at the interface and device current was calculated using a model that takes into account bulk screening of the transient pulse. The voltage dependence of the number of electroporated cells could be explained using a stochastic model for the electroporation kinetics, and the free energy for pore formation was found to be kT at room temperature. With this device, the optimum electroporation conditions can be quickly determined by monitoring the uptake of PI marker dye in situ under the application of millisecond voltage pulses. The electroporation efficiency was also quantified using an ex situ fluorescence-assisted cell sorter, and the morphology of cultured cells was evaluated after the pulsing experiment. Importantly, the efficacy of the developed device was tested independently using two cell lines (C2C12 mouse myoblast cells and yeast cells) as well as in three different electroporation buffers (phosphate buffer saline, electroporation buffer and 10 % glycerol).

Item Type: Journal Article
Publication: EUROPEAN BIOPHYSICS JOURNAL WITH BIOPHYSICS LETTERS
Publisher: SPRINGER
Additional Information: Copy right for this article belongs to the SPRINGER, 233 SPRING ST, NEW YORK, NY 10013 USA
Keywords: Electroporation; Microfluidics; Polymer; Interface; In situ; Fluorescence; Optimization; Model
Department/Centre: Division of Physical & Mathematical Sciences > Physics
Date Deposited: 19 Mar 2015 12:07
Last Modified: 19 Mar 2015 12:07
URI: http://eprints.iisc.ac.in/id/eprint/51039

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