Electrophoresis-assisted single-cell electroporation for efficient intracellular delivery

被引:49
|
作者
Ionescu-Zanetti, Cristian [2 ]
Blatz, Andrew [2 ]
Khine, Michelle [1 ,2 ]
机构
[1] Univ Calif Merced, Sch Engn, Merced, CA 95343 USA
[2] Fluxion Biosci, San Francisco, CA 94158 USA
关键词
single-cell electroporation; electrophoresis; microfluidics;
D O I
10.1007/s10544-007-9115-x
中图分类号
R318 [生物医学工程];
学科分类号
0831 ;
摘要
Single-cell electroporation, in which a focused electric field is applied to permeabilize an individual target cell using relatively low applied voltages, has demonstrated improved cell viability and transfection rates over conventional bulk electroporation set-ups. Here, we introduce a new strategy, in conjunction with single-cell electroporation, to enhance exogenous transport efficiency: electrophoresis delivery of compounds subsequent to electroporation. Electrophoresis is used to assist loading of otherwise impermeable exogenous anionic fluorescent molecules Calcein (Invitrogen, MW=622) and Oregon Green Dextran (OGD, Invitrogen, MW=70,000). For the larger dextran molecules, we demonstrate a protocol of first pre-concentrating at the cell-microfluidic channel interface. Then, the electric field is used to drive these molecules into the cell post-electroporation using 50-200 mV. We demonstrate delivery rate enhancements of more than an order of magnitude using electrophoresis compared to diffusion alone subsequent to electroporation.
引用
收藏
页码:113 / 116
页数:4
相关论文
共 50 条
  • [21] A method of combined single-cell electrophysiology and electroporation
    Graham, Lyle J.
    del Abajo, Ricardo
    Gener, Thomas
    Fernandez, Eduardo
    JOURNAL OF NEUROSCIENCE METHODS, 2007, 160 (01) : 69 - 74
  • [22] A Highly Efficient Method for Single-Cell Electroporation in Mouse Organotypic Hippocampal Slice Culture
    Cheung, Amy
    Futai, Kensuke
    NEUROLOGY, 2021, 96 (15)
  • [23] A highly efficient method for single-cell electroporation in mouse organotypic hippocampal slice culture
    Keener, David G.
    Cheung, Amy
    Futai, Kensuke
    JOURNAL OF NEUROSCIENCE METHODS, 2020, 337
  • [24] Capillary Electrophoresis-Assisted Identification of Peroxyl Radical Generated by Single-Walled Carbon Nanotubes in a Cell-Free System
    Ren, Lei
    Kim, Hong K.
    Zhong, Wenwan
    ANALYTICAL CHEMISTRY, 2009, 81 (13) : 5510 - 5516
  • [25] Sampling techniques for single-cell electrophoresis
    Cecala, Christine
    Sweedler, Jonathan V.
    ANALYST, 2012, 137 (13) : 2922 - 2929
  • [26] Single-cell analysis by capillary electrophoresis
    Stuart, JN
    Sweedler, JV
    ANALYTICAL AND BIOANALYTICAL CHEMISTRY, 2003, 375 (01) : 28 - 29
  • [27] Single-cell analysis by capillary electrophoresis
    Jeffrey N. Stuart
    Jonathan V. Sweedler
    Analytical and Bioanalytical Chemistry, 2003, 375 : 28 - 29
  • [28] Nanostraw-Electroporation System for Highly Efficient Intracellular Delivery and Transfection
    Xie, Xi
    Xu, Alexander M.
    Leal-Ortiz, Sergio
    Cao, Yuhong
    Garner, Craig C.
    Melosh, Nicholas A.
    ACS NANO, 2013, 7 (05) : 4351 - 4358
  • [29] Accurate and efficient intracellular delivery biosensing system by nanostrawed electroporation array
    Fang, Jiaru
    Xu, Jiarong
    Xiang, Yuting
    Li, Hongbo
    Xu, Dongxin
    Xie, Xi
    Hu, Ning
    BIOSENSORS & BIOELECTRONICS, 2021, 194
  • [30] Microfluidic Based Physical Approaches towards Single-Cell Intracellular Delivery and Analysis
    Kaladharan, Kiran
    Kumar, Ashish
    Gupta, Pallavi
    Illath, Kavitha
    Santra, Tuhin Subhra
    Tseng, Fan-Gang
    MICROMACHINES, 2021, 12 (06)