Using a micro electroporation chip to determine the optimal physical parameters in the uptake of biomolecules in HeLa cells

被引:75
作者
He, Huiqi
Chang, Donald C.
Lee, Yi-Kuen
机构
[1] HKUST, Dept Mech Engn, Kowloon, Peoples R China
[2] HKUST, Bioengn Grad Program, Kowloon, Peoples R China
[3] HKUST, Dept Biol, Kowloon, Peoples R China
关键词
micro electroporation; MEMS; 3D electrode; HeLa cells; membrane permeability; phase diagram; size effect;
D O I
10.1016/j.bioelechem.2006.05.008
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
In this study, a new micro electroporation (EP) cell chip with three-dimensional (31)) electrodes was fabricated by means of MEMS technology, and tested on cervical cancer (HeLa) cells. Extensive statistical data of the threshold electric field and pulse duration were determined to construct an EP "phase diagram", which delineates the boundaries for 1) effective EP of five different size molecules and 2) electric cell lysis at the single-cell level. In addition, these boundary curves (i.e., electric field versus pulse duration) were fitted successfully with an exponential function with three constants. We found that, when the molecular size increases, the corresponding electroporation boundary becomes closer to the electric cell lysis boundary. Based on more than 2000single-cell measurements on five different size molecules, the critical size of molecule was found to be approximately 40kDa. Comparing to the traditional instrument, MEMS-based micro electroporation chip can greatly shorten the experimental time. (C) 2006 Elsevier B.V. All rights reserved.
引用
收藏
页码:363 / 368
页数:6
相关论文
共 16 条
[1]  
Chang D. C., 1992, Guide to electroporation and electrofusion., P303
[2]   CELL PORATION AND CELL-FUSION USING AN OSCILLATING ELECTRIC-FIELD [J].
CHANG, DC .
BIOPHYSICAL JOURNAL, 1989, 56 (04) :641-652
[3]   HIGH-EFFICIENCY GENE TRANSFECTION BY ELECTROPORATION USING A RADIOFREQUENCY ELECTRIC-FIELD [J].
CHANG, DC ;
GAO, PQ ;
MAXWELL, BL .
BIOCHIMICA ET BIOPHYSICA ACTA, 1991, 1092 (02) :153-160
[4]  
CHANG DC, 1992, GUIDE ELECTROPORATIO, P1
[5]   Medical applications of electroporation [J].
Dev, SB ;
Rabussay, DP ;
Widera, G ;
Hofmann, GA .
IEEE TRANSACTIONS ON PLASMA SCIENCE, 2000, 28 (01) :206-223
[6]   Enhancement of cytotoxicity by electropermeabilization: an improved method for screening drugs [J].
Gehl, J ;
Skovsgaard, T ;
Mir, LM .
ANTI-CANCER DRUGS, 1998, 9 (04) :319-325
[7]  
HE H, 2005, BIOELECTROCHEMISTRY, V68, P91
[8]   Microfabricated electroporation chip for single cell membrane permeabilization [J].
Huang, Y ;
Rubinsky, B .
SENSORS AND ACTUATORS A-PHYSICAL, 2001, 89 (03) :242-249
[9]   A single cell electroporation chip [J].
Khine, M ;
Lau, A ;
Ionescu-Zanetti, C ;
Seo, J ;
Lee, LP .
LAB ON A CHIP, 2005, 5 (01) :38-43
[10]  
LIANG H, 1998, BIOTECNIQUES, V6, P550