高时间分辨率下观察到的巨型囊泡的电变形和穿孔

Electro-deformation and poration of giant vesicles viewed with high temporal resolution.

作者信息

Riske Karin A, Dimova Rumiana

机构信息

Max Planck Institute of Colloids and Interfaces, 14476 Golm, Germany.

出版信息

Biophys J. 2005 Feb;88(2):1143-55. doi: 10.1529/biophysj.104.050310. Epub 2004 Dec 13.

Abstract

Fast digital imaging was used to study the deformation and poration of giant unilamellar vesicles subjected to electric pulses. For the first time the dynamics of response and relaxation of the membrane at micron-scale level is revealed at a time resolution of 30 micros. Above a critical transmembrane potential the lipid bilayer ruptures. Formation of macropores (diameter approximately 2 microm) with pore lifetime of approximately 10 ms has been detected. The pore lifetime has been interpreted as interplay between the pore edge tension and the membrane viscosity. The reported data, covering six decades of time, show the following regimes in the relaxation dynamics of the membrane. Tensed vesicles first relax to release the acquired stress due to stretching, approximately 100 micros. In the case of poration, membrane resealing occurs with a characteristic time of approximately 10 ms. Finally, for vesicles with excess area an additional slow regime was observed, approximately 1 s, which we associate with relaxation of membrane curvature. Dimensional analysis can reasonably well explain the corresponding characteristic timescales. Being performed on cell-sized giant unilamellar vesicles, this study brings insight to cell electroporation. The latter is widely used for gene transfection and drug transport across the membrane where processes occurring at different timescales may influence the efficiency.

摘要

利用快速数字成像技术研究了巨型单层囊泡在电脉冲作用下的变形和穿孔情况。首次以30微秒的时间分辨率揭示了微米尺度水平下膜的响应和弛豫动力学。高于临界跨膜电位时,脂质双分子层会破裂。已检测到形成了直径约2微米、孔寿命约为10毫秒的大孔。孔寿命被解释为孔边缘张力与膜粘度之间的相互作用。所报告的数据涵盖六个数量级的时间,显示了膜弛豫动力学中的以下几种情况。紧绷的囊泡首先弛豫以释放因拉伸而获得的应力,约100微秒。在穿孔的情况下,膜重新封闭的特征时间约为10毫秒。最后,对于具有多余面积的囊泡,观察到一种额外的缓慢情况,约1秒,我们将其与膜曲率的弛豫联系起来。量纲分析能够较好地解释相应的特征时间尺度。这项在细胞大小的巨型单层囊泡上进行的研究为细胞电穿孔提供了深入见解。后者广泛用于基因转染和药物跨膜运输,其中在不同时间尺度上发生的过程可能会影响效率。

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