LAURDAN since Weber: The Quest for Visualizing Membrane Heterogeneity.
LAURDAN since Weber: The Quest for Visualizing Membrane Heterogeneity.
复制标题
自韦伯以来的劳尔丹：可视化膜异质性的探索。
DOI：
10.1021/acs.accounts.0c00687
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发表时间：
2021-02-16
影响因子：
18.3
通讯作者：
Sánchez SA
中科院分区：
文献类型：
--
作者：
Gunther G;Malacrida L;Jameson DM;Gratton E;Sánchez SA
任何研究分子与脂质组装体相互作用的化学家最终都会遇到膜双层异质性的问题，最终可能会遇到天然膜的异质性。在人工双层中，非均质性是由纳米和微米范围内的相分离定义的。在生物双层中，异质性被认为是在小的(10-200 nm)甾醇和鞘磷脂富含的异质和高度动态的结构域的背景下进行的。有几种技术可以用来评估生物系统中的膜的异质性。我们的方法是使用浸泡在双层中的荧光报告分子，通过其光谱性质的变化来感知膜的物理化学方面。这种染料与显微镜和波动技术相结合，可以在不同的时间和空间水平上提供关于膜的异质性的信息：从平均流动性到纳米结构域的数量和扩散系数。Laurdan(6-十二酰基-2-二甲氨基)萘是Gregorio Weber于1979年设计合成的一种用于研究偶极弛豫现象的荧光探针。当Laurdan处于流体或凝胶相时所观察到的光谱位移使得该技术能够在膜动力学领域中使用。光谱位移的量化首先是通过试管中的广义偏振(GP)函数来解决的，即两个波长的强度差除以它们的总和的比率。1997年，第一次在显微镜下进行GP测量，增加了空间分辨率，并允许可视化脂质体和细胞中的脂类分离。当在显微镜下进行Laurdan荧光寿命测量时，对膜的异质性有了新的展望。双通道寿命成像提供了关于膜极性和偶极驰豫的信息(这两个参数是导致Laurdan光谱漂移的两个参数)，相量分析的应用使我们能够逐个像素地了解膜中的这两个参数。为了提高时间分辨率，将Laurdan GP与涨落相关光谱(FCS)相结合，记录了生物膜中纳米高填充结构的运动性。最近，将相量分析应用于Laurdan标记的膜的光谱图像，使我们能够逐个像素地研究图像中的全光谱。所有这些方法，使用Laurdan，提供了根据所提出的问题来解决膜的不同性质的可能性。在这篇文章中，我们将集中讨论不同方法的原理、优点和局限性，以引导读者选择最适合他们研究的技术。
Any chemist studying the interaction of molecules with lipid assemblies will eventually be confronted by the topic of membrane bilayer heterogeneity and may ultimately encounter the heterogeneity of natural membranes. In artificial bilayers, heterogeneity is defined by phase segregation that can be in the nano- and micrometer range. In biological bilayers, heterogeneity is considered in the context of small (10–200 nm) sterol and sphingolipid-enriched heterogeneous and highly dynamic domains. Several techniques can be used to assess membrane heterogeneity in living systems. Our approach is to use a fluorescent reporter molecule immersed in the bilayer, which, by changes in its spectroscopic properties, senses physical-chemistry aspects of the membrane. This dye in combination with microscopy and fluctuation techniques can give information about membrane heterogeneity at different temporal and spatial levels: going from average fluidity to number and diffusion coefficient of nanodomains. LAURDAN (6-dodecanoyl-2-(dimethylamino) naphthalene), is a fluorescent probe designed and synthesized in 1979 by Gregorio Weber with the purpose to study the phenomenon of dipolar relaxation. The spectral displacement observed when LAURDAN is either in fluid or gel phase permitted the use of the technique in the field of membrane dynamics. The quantitation of the spectral displacement was first addressed by the generalized polarization (GP) function in the cuvette, a ratio of the difference in intensity at two wavelengths divided by their sum. In 1997, GP measurements were done for the first time in the microscope, adding to the technique the spatial resolution and allowing the visualization of lipid segregation both in liposomes and cells. A new prospective to the membrane heterogeneity was obtained when LAURDAN fluorescent lifetime measurements were done in the microscope. Two channel lifetime imaging provides information on membrane polarity and dipole relaxation (the two parameters responsible for the spectral shift of LAURDAN), and the application of phasor analysis allows pixel by pixel understanding of these two parameters in the membrane. To increase temporal resolution, LAURDAN GP was combined with fluctuation correlation spectroscopy (FCS) and the motility of nanometric highly packed structures in biological membranes was registered. Lately the application of phasor analysis to spectral images from membranes labeled with LAURDAN allows us to study the full spectra pixel by pixel in an image. All these methodologies, using LAURDAN, offer the possibility to address different properties of membranes depending on the question being asked. In this Account, we will focus on the principles, advantages, and limitations of different approaches to orient the reader to select the most appropriate technique for their research.
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影响因子：
3.8
作者：
Fereidouni, Farzad;Bader, Arjen N.;Gerritsen, Hans C.
通讯作者：
Gerritsen, Hans C.
影响因子：
3.4
作者：
Dietrich, C;Bagatolli, LA;Gratton, E
通讯作者：
Gratton, E
影响因子：
4.8
作者：
Fernández-Pérez EJ;Sepúlveda FJ;Peters C;Bascuñán D;Riffo-Lepe NO;González-Sanmiguel J;Sánchez SA;Peoples RW;Vicente B;Aguayo LG
通讯作者：
Aguayo LG
影响因子：
3.4
作者：
Celli, A.;Sanchez, S.;Mauro, T.
通讯作者：
Mauro, T.
影响因子：
2.6
作者：
Bonaventura, Gabriele;Barcellona, Maria Luisa;Golfetto, Ottavia;Nourse, Jamison L.;Flanagan, Lisa A.;Gratton, Enrico
通讯作者：
Gratton, Enrico