Regional electroporation of single cardiac myocytes in a focused electric field

Klauke, N., Smith, G. and Cooper, J.M. (2010) Regional electroporation of single cardiac myocytes in a focused electric field. Analytical Chemistry, 82(2), pp. 585-592. (doi:10.1021/ac901886j)

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Publisher's URL: http://dx.doi.org/10.1021/ac901886j

Abstract

There is now a significant interest in being able to locate single cells within geometrically defined regions of a microfluidic chip and to gain intracellular access through the local electroporation of the cell membrane. This paper describes the microfabrication of electroporation devices which can enable the regional electroporation of adult ventricular myocytes, in order to lower the local electrical resistance of the cell membrane. Initially three different devices, designed to suit the characteristic geometry of the cardiomyocyte, were investigated (all three designs serve to focus the electric field to selected regions of the cell). We demonstrate that one of these three devices revealed the sequence of cellular responses to field strengths of increasing magnitudes, namely, cell contraction, hypercontraction, and lysis. This same device required a reduced threshold voltage for each of these events, including in particular membrane breakdown. We were not only able to show the gradual regional increase in the electric conductivity of the cell membrane but were also able to avoid changes in the local intra- and extracellular pH (by preventing the local generation of protons at the electrode surface, as a consequence of the reduced threshold voltage). The paper provides evidence for new strategies for achieving robust and reproducible regional electroporation, a technique which, in future, may be used for the insertion of large molecular weight molecules (including genes) as well as for on-chip voltage clamping of the primary adult cardiomyocyte.

Item Type:Articles
Keywords:Cardiomyocyte, cardiomyocytes, cell electroporation, cells, chip, contraction, design, device, devices, excitation, gene, generation, integrity, intracellular ph, mammalian-cells, microfabrication, recordings, single cardiac myocytes, single cells, stimulation, threshold, ventricular myocytes
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Klauke, Dr Norbert and Smith, Professor Godfrey and Cooper, Professor Jonathan
Authors: Klauke, N., Smith, G., and Cooper, J.M.
Subjects:Q Science > QH Natural history > QH345 Biochemistry
College/School:College of Medical Veterinary and Life Sciences > Institute of Cardiovascular and Medical Sciences
College of Medical Veterinary and Life Sciences
College of Science and Engineering > School of Engineering > Electronics and Nanoscale Engineering
College of Science and Engineering > School of Engineering > Biomedical Engineering
Journal Name:Analytical Chemistry
ISSN:0003-2700
ISSN (Online):1520-6882
Published Online:18 December 2009

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
393961Electrophysiology of ventricular myofybroblasts within myocardial infarction scars of rabbit heartsGodfrey SmithBritish Heart Foundation (BHF)PG/05/032Institute of Cardiovascular and Medical Sciences