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荧光相关和寿命相关光谱学应用于细胞膜支持脂质双层模型的研究。

Fluorescence correlation and lifetime correlation spectroscopy applied to the study of supported lipid bilayer models of the cell membrane.

作者信息

Basit Hajra, Lopez Sergio G, Keyes Tia E

机构信息

School of Chemical Sciences, National Biophotonics and Imaging Platform, Dublin City University, Glasnevin, Dublin 9, Ireland.

School of Chemical Sciences, National Biophotonics and Imaging Platform, Dublin City University, Glasnevin, Dublin 9, Ireland.

出版信息

Methods. 2014 Jul 1;68(2):286-99. doi: 10.1016/j.ymeth.2014.02.005. Epub 2014 Feb 20.

Abstract

Supported Lipid Bilayers (SLBs) are versatile models capable of mimicking some of the key properties of the cell membrane, including for example lipid fluidity, domain formation and protein support, without the challenging complexity of the real biological system. This is important both from the perspective of understanding the behaviour and role of the lipid membrane in cell structure and signalling, as well as in development of applications of lipid membranes across domains as diverse as sensing and drug delivery. Lipid and protein diffusion within the membrane is vital to its function and there are several key experimental methods used to study membrane dynamics. Amongst the optical methods are Fluorescence Recovery After Photobleaching (FRAP), single particle tracking and Fluorescence Correlation (and Fluorescence Lifetime Correlation) Spectroscopy (FCS/FLCS). Each of these methods can provide different and often complementary perspectives on the dynamics of the fluid membrane. Although FCS is well established, FLCS is a relatively new technique and both methods have undergone a number of extensions in recent years which improve their precision and accuracy in studying supported lipid bilayers, most notably z-scan methods. This short review focusses on FCS and FLCS and their recent applications, specifically to artificial lipid bilayer studies addressing key issues of cell membrane behaviour.

摘要

支持脂质双分子层(SLBs)是通用模型,能够模拟细胞膜的一些关键特性,例如脂质流动性、结构域形成和蛋白质支持等,而无需面对真实生物系统中具有挑战性的复杂性。这从理解脂质膜在细胞结构和信号传导中的行为及作用,以及脂质膜在传感和药物递送等不同领域的应用开发角度来看都很重要。膜内脂质和蛋白质的扩散对其功能至关重要,有几种关键的实验方法用于研究膜动力学。光学方法包括光漂白后荧光恢复(FRAP)、单粒子追踪以及荧光相关(和荧光寿命相关)光谱法(FCS/FLCS)。这些方法中的每一种都能为流体膜的动力学提供不同且往往互补的视角。虽然FCS已得到广泛应用,但FLCS是一项相对较新的技术,近年来这两种方法都有了一些扩展,提高了它们在研究支持脂质双分子层时的精度和准确性,最显著的是z扫描方法。这篇简短的综述聚焦于FCS和FLCS及其近期应用,特别是针对解决细胞膜行为关键问题的人工脂质双分子层研究。

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