Fluorescence correlation spectroscopy with single-molecule sensitivity on cell and model membranes
Fluorescence correlation spectroscopy with single-molecule sensitivity on cell and model membranes
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DOI：
10.1002/(sici)1097-0320(19990701)36:3
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发表时间：
1999-07-01
期刊：
影响因子：
--
通讯作者：
Webb, WW
中科院分区：
文献类型：
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作者：
Schwille, P;Korlach, J;Webb, WW
我们报告的成功应用荧光相关光谱（FCS）的分析单荧光标记的脂质类似物的分子扩散横向脂质双层，作为例证的时间轨迹的荧光猝发的单个分子进入和离开的激发区。在大鼠嗜碱性白血病细胞膜上进行的脂质探针FCS测量显示，二维布朗运动与单一的均匀扩散常数的偏差。以单层巨囊泡为模型系统，研究了荧光脂质类似物在均相和混合脂相中的扩散特性。将细胞膜扩散的结果与模型系统的结果进行比较，提出了对观察结果的可能解释。(a)异常亚扩散，其中与不同的移动的分子的瞬逝吸引相互作用改变了扩散统计;（B）或者，探针分子位于亚微观尺寸的微区中，可能在异质膜相中。Cytometry 36：176-182，1999. (C)1999 Wiley-Liss，Inc.
We report on the successful application of fluorescence correlation spectroscopy (FCS) to the analysis of single fluorescently labeled lipid analogue molecules diffusing laterally in lipid bilayers, as exemplified by time traces of fluorescence bursts of individual molecules entering and leaving the excitation area. FCS measurements performed on lipid probes in rat basophilic leukemia cell membranes showed deviations from two-dimensional Brownian motion with a single uniform diffusion constant. Giant unilamellar vesicles were employed as model systems to characterize diffusion of fluorescent lipid analogues in both homogeneous and mixed lipid phases with diffusion heterogeneity. Comparing the results of cell membrane diffusion with the findings on the model systems suggests possible explanations for the observations. (a) anomalous subdiffusion in which evanescent attractive interactions with disparate mobile molecules modifies the diffusion statistics; (b) alternatively, probe molecules are localized in microdomains of submicroscopic size, possibly in heterogeneous membrane phases. Cytometry 36:176-182, 1999. (C) 1999 Wiley-Liss, Inc.