help button home button Biophys. J.
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Ryttsén, F.
Right arrow Articles by Orwar, O.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Ryttsén, F.
Right arrow Articles by Orwar, O.

Biophys J, October 2000, p. 1993-2001, Vol. 79, No. 4

Characterization of Single-Cell Electroporation by Using Patch-Clamp and Fluorescence Microscopy

Frida Ryttsén,* Cecilia Farre,* Carrie Brennan,dagger Stephen G. Weber,dagger Kerstin Nolkrantz,* Kent Jardemark,* Daniel T. Chiu,* and Owe Orwar*

 *Department of Chemistry, Göteborg University, Göteborg SE-412 96, Sweden and  dagger Department of Chemistry, University of Pittsburgh, Pittsburgh, Pennsylvania, 15260 USA

Electroporation of single NG108-15 cells with carbon-fiber microelectrodes was characterized by patch-clamp recordings and fluorescence microscopy. To minimize adverse capacitive charging effects, the patch-clamp pipette was sealed on the cell at a 90o angle with respect to the microelectrodes where the applied potential reaches a minimum. From transmembrane current responses, we determined the electric field strengths necessary for ion-permeable pore formation and investigated the kinetics of pore opening and closing as well as pore open times. From both patch-clamp and fluorescence microscopy experiments, the threshold transmembrane potentials for dielectric breakdown of NG108-15 cells, using 1-ms rectangular waveform pulses, was ~250 mV. The electroporation pulse preceded pore formation, and analyte entry into the cells was dictated by concentration, and membrane resting potential driving forces. By stepwise moving a cell out of the focused field while measuring the transmembrane current response during a supramaximal pulse, we show that cells at a distance of ~30 µm from the focused field were not permeabilized.

Biophys J, October 2000, p. 1993-2001, Vol. 79, No. 4
© 2000 by the Biophysical Society   0006-3495/00/10/1993/09  $2.00


This article has been cited by other articles:


Home page
 Biophys. JHome page
A. Bruckbauer, P. James, D. Zhou, J. W. Yoon, D. Excell, Y. Korchev, R. Jones, and D. Klenerman
Nanopipette Delivery of Individual Molecules to Cellular Compartments for Single-Molecule Fluorescence Tracking
Biophys. J., November 1, 2007; 93(9): 3120 - 3131.
[Abstract] [Full Text] [PDF]

Home page
 Biophys. JHome page
I. Zudans, A. Agarwal, O. Orwar, and S. G. Weber
Numerical Calculations of Single-Cell Electroporation with an Electrolyte-Filled Capillary
Biophys. J., May 15, 2007; 92(10): 3696 - 3705.
[Abstract] [Full Text] [PDF]

Home page
 Biophys. JHome page
M. Pavlin, M. Kanduser, M. Rebersek, G. Pucihar, F. X. Hart, R. Magjarevic, and D. Miklavcic
Effect of Cell Electroporation on the Conductivity of a Cell Suspension
Biophys. J., June 1, 2005; 88(6): 4378 - 4390.
[Abstract] [Full Text] [PDF]

Home page
 Biophys. JHome page
M. Pavlin and D. Miklavcic
Effective Conductivity of a Suspension of Permeabilized Cells: A Theoretical Analysis
Biophys. J., August 1, 2003; 85(2): 719 - 729.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Copyright © 2000 by the Biophysical Society.