Effect of cell size and shape on single-cell electroporation

被引:87
作者
Agarwal, Aparna
Zudans, Imants
Weber, Emily A.
Olofsson, Jessica
Orwar, Owe
Weber, Stephen G. [1 ]
机构
[1] Univ Pittsburgh, Dept Chem, Pittsburgh, PA 15260 USA
[2] Johns Hopkins Univ, Bloomberg Sch Publ Hlth, Baltimore, MD 21205 USA
[3] Chalmers, Dept Phys Chem, SE-41296 Gothenburg, Sweden
关键词
D O I
10.1021/ac062049e
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
Single-cell electroporation was performed using electrolyte-filled capillaries on fluorescently labeled A549 cells. Cells were exposed to brief pulses (50-300 ms) at various cell-capillary tip distances. Cell viability and electroporation success were measured. In order to understand the variability in single-cell electroporation, logistic regression was used to determine whether the probabilities of cell survival and electroporation depend on experimental conditions and cell properties. Both experimental conditions and cell properties (size and shape) have a significant effect on the outcome. Finite element simulations were used to compare bulk electroporation to single-cell electroporation in terms of cell size and shape. Cells are more readily permeabilized and are more likely to survive if they are large and hemispherical as opposed to small and ellipsoidal with a high aspect ratio. The dependence of the maximum transmembrane potential across the cell membrane on cell size is much weaker than it is for bulk electroporation. Observed survival probabilities are related to the calculated fraction of the cell's surface area that is electroporated. Observed success of electroporation is related to the maximum transmembrane potential achieved.
引用
收藏
页码:3589 / 3596
页数:8
相关论文
共 43 条
[1]   Simultaneous maximization of cell permeabilization and viability in single-cell electroporation using an electrolyte-filled capillary [J].
Agarwal, Aparna ;
Zudans, Imants ;
Orwar, Owe ;
Weber, Stephen G. .
ANALYTICAL CHEMISTRY, 2007, 79 (01) :161-167
[2]   Microtechnologies and nanotechnologies for single-cell analysis [J].
Andersson, H ;
van den Berg, A .
CURRENT OPINION IN BIOTECHNOLOGY, 2004, 15 (01) :44-49
[3]   The generation of reactive-oxygen species associated with long-lasting pulse-induced electropermeabilisation of mammalian cells is based on a non-destructive alteration of the plasma membrane [J].
Bonnafous, P ;
Vernhes, MC ;
Teissié, J ;
Gabriel, B .
BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES, 1999, 1461 (01) :123-134
[4]   Nanoscale electrochemical probes for single cell analysis [J].
Fasching, R. J. ;
Bai, S. -J. ;
Fabian, T. ;
Prinz, F. B. .
MICROELECTRONIC ENGINEERING, 2006, 83 (4-9) :1638-1641
[5]   Electric field-induced cell membrane permeabilization and gene transfer:: Theory and experiments [J].
Faurie, C ;
Golzio, M ;
Phez, E ;
Teissié, J ;
Rols, MP .
ENGINEERING IN LIFE SCIENCES, 2005, 5 (02) :179-186
[6]   Electroporation of cells in microfluidic devices: a review [J].
Fox, M. B. ;
Esveld, D. C. ;
Valero, A. ;
Luttge, R. ;
Mastwijk, H. C. ;
Bartels, P. V. ;
van den Berg, A. ;
Boom, R. M. .
ANALYTICAL AND BIOANALYTICAL CHEMISTRY, 2006, 385 (03) :474-485
[7]   CONTROL BY ELECTRICAL PARAMETERS OF SHORT-TERM AND LONG-TERM CELL-DEATH RESULTING FROM ELECTROPERMEABILIZATION OF CHINESE-HAMSTER OVARY CELLS [J].
GABRIEL, B ;
TEISSIE, J .
BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR CELL RESEARCH, 1995, 1266 (02) :171-178
[8]   High-throughput single-cell analysis for enzyme activity without cytolysis [J].
Gao, Ning ;
Wang, Wenlei ;
Zhang, Xiaoli ;
Jin, Wenrui ;
Yin, Xuefeng ;
Fang, Zhaolun .
ANALYTICAL CHEMISTRY, 2006, 78 (09) :3213-3220
[9]  
Gift EA, 2000, CYTOMETRY, V39, P243
[10]   A polarization model overcoming the geometric restrictions of the laplace solution for spheroidal cells: Obtaining new equations for field-induced forces and transmembrane potential [J].
Gimsa, J ;
Wachner, D .
BIOPHYSICAL JOURNAL, 1999, 77 (03) :1316-1326