Comparison of optical saturation effects in conventional and dual-focus fluorescence correlation spectroscopy

被引:31
作者
Loman, Anastasia [1 ]
Dertinger, Thomas [2 ]
Koberling, Felix [3 ]
Enderlein, Joerg [1 ]
机构
[1] Univ Tubingen, Inst Phys & Theoret Chem, D-72076 Tubingen, Germany
[2] Univ Calif Los Angeles, Dept Chem & Biochem, Los Angeles, CA 90095 USA
[3] PicoQuant GmbH, D-12961 Berlin, Germany
关键词
D O I
10.1016/j.cplett.2008.05.018
中图分类号
O64 [物理化学(理论化学)、化学物理学];
学科分类号
070304 ; 081704 ;
摘要
We present diffusion measurements of the fluorescent dye Cy5 in aqueous solutions using conventional fluorescence correlation spectroscopy (FCS) and the recently introduced dual-focus fluorescence correlation spectroscopy (2fFCS). We study the sensitivity of both methods with respect to excitation intensity. Due to the light-driven transitions of Cy5 between fluorescent and non-fluorescent states, conventional FCS shows a strong dependence of the apparent diffusion coefficient on excitation intensity, whereas 2fFCS is virtually free from that artifact. Moreover, 2fFCS allows us to obtain the first precisely measured absolute value for the diffusion coefficient of Cy5. (C) 2008 Elsevier B.V. All rights reserved.
引用
收藏
页码:18 / 21
页数:4
相关论文
共 25 条
[1]   How to determine diffusion coefficients in planar phospholipid systems by confocal fluorescence correlation spectroscopy [J].
Benda, A ;
Benes, M ;
Marecek, V ;
Lhotsky, A ;
Hermens, WT ;
Hof, M .
LANGMUIR, 2003, 19 (10) :4120-4126
[2]   Excitation saturation in two-photon fluorescence correlation spectroscopy [J].
Berland, K ;
Shen, GQ .
APPLIED OPTICS, 2003, 42 (27) :5566-5576
[3]   Two-focus fluorescence correlation spectroscopy: A new tool for accurate and absolute diffusion measurements [J].
Dertinger, Thomas ;
Pacheco, Victor ;
von der Hocht, Iris ;
Hartmann, Rudolf ;
Gregor, Ingo ;
Enderlein, Joerg .
CHEMPHYSCHEM, 2007, 8 (03) :433-443
[4]   FLUORESCENCE CORRELATION SPECTROSCOPY .1. CONCEPTUAL BASIS AND THEORY [J].
ELSON, EL ;
MAGDE, D .
BIOPOLYMERS, 1974, 13 (01) :1-27
[5]   Performance of fluorescence correlation spectroscopy for measuring diffusion and concentration [J].
Enderlein, J ;
Gregor, I ;
Patra, D ;
Dertinger, T ;
Kaupp, UB .
CHEMPHYSCHEM, 2005, 6 (11) :2324-2336
[6]   Art and artefacts of fluorescence correlation spectroscopy [J].
Enderlein, J ;
Gregor, I ;
Patra, D ;
Fitter, J .
CURRENT PHARMACEUTICAL BIOTECHNOLOGY, 2004, 5 (02) :155-161
[7]  
ENDERLEIN J, 2005, REV SCI INSTRUM, V76
[8]   Fluorescence correlation spectroscopy of molecular motions and kinetics [J].
Gösch, M ;
Rigler, R .
ADVANCED DRUG DELIVERY REVIEWS, 2005, 57 (01) :169-190
[9]   Optical saturation in fluorescence correlation spectroscopy under continuous-wave and pulsed excitation [J].
Gregor, I ;
Patra, D ;
Enderlein, J .
CHEMPHYSCHEM, 2005, 6 (01) :164-170
[10]   Fluorescence correlation spectroscopy: Novel variations of an established technique [J].
Haustein, Elke ;
Schwille, Petra .
ANNUAL REVIEW OF BIOPHYSICS AND BIOMOLECULAR STRUCTURE, 2007, 36 :151-169