Total Internal Reflection Two-Dimensional Fluorescence Lifetime Correlation Spectroscopy.
Total Internal Reflection Two-Dimensional Fluorescence Lifetime Correlation Spectroscopy.
复制标题
全内反射二维荧光寿命相关光谱。
DOI：
10.1021/acs.jpcb.8b01176
复制
发表时间：
2018
影响因子：
3.3
通讯作者：
S. Yamaguchi
中科院分区：
文献类型：
--
作者：
T. Otosu;S. Yamaguchi
荧光寿命相关分析正以前所未有的检测灵敏度和时间分辨率成为了解生物分子构象异质性及其动力学的有力工具。然而，它在生物膜研究中的应用非常有限。在这里，我们报道了二维荧光寿命相关光谱（2D FLCS）与全内反射（TIR）显微镜（TIR 2D-FLCS）的结合。TIR显微镜的高深度分辨率和2D FLCS的物种特异性相关性分析使我们有机会选择性地分析支持的脂质双分子层内或上的分子，这是在玻璃表面形成的模型生物膜。本研究中进行的可行性实验清楚地表明，TIR 2D-FLCS具有选择性分析膜上外周结合的蛋白质在体相中存在大量未结合分子时的扩散和构象动力学的潜力。
Fluorescence lifetime correlation analysis is becoming a powerful tool to understand the conformational heterogeneity of biomolecules and their dynamics with an unprecedented detection sensitivity and time resolution. However, its application to the study of biomembranes is very limited. Here, we report on two-dimensional fluorescence lifetime correlation spectroscopy (2D FLCS) in combination with total internal reflection (TIR) microscopy (TIR 2D-FLCS). High depth resolution in TIR microscopy and species-specific correlation analysis in 2D FLCS give us the opportunity to selectively analyze molecules in or on a supported lipid bilayer, a model biomembrane formed on the glass surface. Feasibility experiments performed in this study clearly demonstrated that TIR 2D-FLCS has a potential to selectively analyze the diffusion and the conformational dynamics of proteins peripherally bound on the membrane in the presence of substantial amounts of unbound molecules in the bulk phase.