Simultaneous Surface-Near and Solution Fluorescence Correlation Spectroscopy

被引：0
作者：
Christian M. Winterflood
Stefan Seeger
机构：
[1] King’s College London,Randall Division of Cell and Molecular Biophysics
[2] University of Zurich,Department of Chemistry
来源：
Journal of Fluorescence | 2016年 / 26卷
关键词：
Fluorescence correlation spectroscopy; Supercritical angle fluorescence; Undercritical angle fluorescence; Surface-selective; Near-field; Far-field;
D O I：
暂无
中图分类号：
学科分类号：
摘要：
We report the first simultaneous measurement of surface-confined and solution fluorescence correlation spectroscopy (FCS). We use an optical configuration for tightly focused excitation and separate detection of light emitted below (undercritical angle fluorescence, UAF) and above (supercritical angle fluorescence, SAF) the critical angle of total internal reflection of the coverslip/sample interface. This creates two laterally coincident detection volumes which differ in their axial extent. While detection of far-field UAF emission producesa standard confocal volume, near-field-mediated SAF produces a highly surface-confined detection volume at the coverslip/sample interface which extends only ~200 nm into the sample. A characterization of the two detection volumes by FCS of free diffusion is presented and compared with analytical models and simulations. The presented FCS technique allows to determine bulk solution concentrations and surface-near concentrations at the same time.
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页码：753 / 756
页数：3
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