Quantified femtosecond laser based opto-perforation of living GFSHR-17 and MTH53a cells

被引：87

作者：

Baumgart, J. ^{[1
]}

Bintig, W. ^{[2
]}

Ngezahayo, A. ^{[2
]}

Willenbrock, S. ^{[3
]}

Escobar, H. Murua ^{[3
]}

Ertmer, W. ^{[4
]}

Lubatschowski, H. ^{[1
]}

Heisterkamp, A. ^{[1
]}

机构：

[1] Laser Zentrum Hannover eV, Hannover, Germany

[2] Leibniz Univ Hannover, Inst Biophys, Hannover, Germany

[3] Univ Vet Med, Small Anim Clin, Hannover, Germany

[4] Leibniz Univ Hannover, Inst Quantum Phys, Hannover, Germany

来源：

OPTICS EXPRESS | 2008年 / 16卷 / 05期

关键词：

D O I：

10.1364/OE.16.003021

中图分类号：

O43 [光学];

学科分类号：

070207 ; 0803 ;

摘要：

Opto-perforation is an interesting alternative to conventional techniques for gene transfer into living cells. The cell membrane is perforated by femtosecond (fs) laser pulses, in order to induce an uptake of macromolecules e. g. DNA. In this study, we successfully transfected a canine cell line ( MTH53a) with GFP vector or a vector coding for a GFP-HMGB1 fusion protein. The transfected cells were observed 48 hours after treatment and they were not showing any signs of apoptosis or necrosis. Based on simultaneously measured membrane potential changes during the perforation, we were able to calculate and experimentally verify that the relative volume exchanged is 0.4 times the total cell volume. Thus, for first time a quantitative predication of the amount of uptaken molecules and therefore a quantification of the transfection is possible. Additionally, this method offers new high efficient possibilities for critical transfection approaches involving special cell types, e. g. primary and stem cells. (C) 2008 Optical Society of America.

引用

页码：3021 / 3031

页数：11

共 17 条

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