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Numerical Fluorescence Correlation Spectroscopy for the Analysis of Molecular Dynamics under Nonstandard Conditions

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Michael J. Culbertson,^† Joshua T. B. Williams,^† Wayland W. L. Cheng,^† Dee Ann Stults,^† Emily R. Wiebracht,^† John J. Kasianowicz,^‡ and Daniel L. Burden*^†

Chemistry Department, Wheaton College, Wheaton, Illinois 60187, and EEEL Division, National Institute of Standards and Technology, Gaithersburg, Maryland 20899

Anal. Chem., 2007, 79 (11), pp 4031–4039

DOI: 10.1021/ac062013m

Publication Date (Web): April 21, 2007

Section:

Optical, Electron, and Mass Spectroscopy and Other Related Properties

Abstract

The suitability of mathematical models used to extract kinetic information from correlated data constitutes a significant issue in fluorescence correlation spectroscopy (FCS). Standard FCS equations are derived from a simple Gaussian approximation of the optical detection volume, but some investigations have suggested this traditional practice can lead to inaccurate and misleading conclusions under many experimental circumstances, particularly those encountered in one-photon confocal measurements. Furthermore, analytical models cannot be derived for all measurement scenarios. We describe a novel numerical approach to FCS that circumvents conventional analytical models, enabling meaningful analyses even under extraordinarily unusual measurement conditions. Numerical fluorescence correlation spectroscopy (NFCS) involves quantitatively matching experimental correlation curves with synthetic curves generated via diffusion simulation or direct calculation based on an experimentally determined 3D map of the detection volume. Model parameters are adjusted iteratively to minimize the residual differences between synthetic and experimental correlation curves. In order to reduce analysis time, we distribute calculations across a network of processors. As an example of this new approach, we demonstrate that synthetic autocorrelation curves correspond well with experimental data and that NFCS diffusion measurements of Rhodamine B remain constant, regardless of the distortion present in a confocal detection volume.

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This article has been cited by 5 ACS Journal articles (5 most recent appear below).

Information Content in Fluorescence Correlation Spectroscopy: Binary Mixtures and Detection Volume Distortion
Jonathan D. Lam, Michael J. Culbertson, Nathan P. Skinner, Zachary J. Barton, and Daniel L. Burden
Analytical Chemistry2011 Article ASAP
- Information Content in Fluorescence Correlation Spectroscopy: Binary Mixtures and Detection Volume Distortion
  Jonathan D. Lam, Michael J. Culbertson, Nathan P. Skinner, Zachary J. Barton, and Daniel L. Burden
  Analytical Chemistry2011 Article ASAP
  When properly implemented, fluorescence correlation spectroscopy (FCS) reveals numerous static and dynamic properties of molecules in solution. However, complications arise whenever the measurement scenario is complex. Specific limitations occur when the ...
  Abstract | Full Text HTML | Hi-Res PDF | PDF w/ Links
FRET Fluctuation Spectroscopy of Diffusing Biopolymers: Contributions of Conformational Dynamics and Translational Diffusion
Kaushik Gurunathan and Marcia Levitus
The Journal of Physical Chemistry B2010 114 (2), 980-986
- FRET Fluctuation Spectroscopy of Diffusing Biopolymers: Contributions of Conformational Dynamics and Translational Diffusion
  Kaushik Gurunathan and Marcia Levitus
  The Journal of Physical Chemistry B2010 114 (2), 980-986
  The use of fluorescence correlation spectroscopy (FCS) to study conformational dynamics in diffusing biopolymers requires that the contributions to the signal due to translational diffusion are separated from those due to conformational dynamics. A simple ...
  Abstract | Full Text HTML | Hi-Res PDF | PDF w/ Links
Effect of Cross-Linking on the Diffusion of Water, Ions, and Small Molecules in Hydrogels
Yanbin Wu, Sony Joseph and N. R. Aluru
The Journal of Physical Chemistry B2009 113 (11), 3512-3520
- Effect of Cross-Linking on the Diffusion of Water, Ions, and Small Molecules in Hydrogels
  Yanbin Wu, Sony Joseph and N. R. Aluru
  The Journal of Physical Chemistry B2009 113 (11), 3512-3520
  The present study reports on molecular dynamics investigations of chemically cross-linked poly(ethylene glycol) hydrogels with the aim of exploring the diffusion properties of water, ions, and rhodamine within the polymer at the molecular level. The water ...
  Abstract | Full Text HTML | Hi-Res PDF | PDF w/ Links
Fluorescence Correlation Spectroscopy: Criteria for Analysis in Complex Systems
Alexei Tcherniak, Carmen Reznik, Stephan Link and Christy F. Landes
Analytical Chemistry2009 81 (2), 746-754
- Fluorescence Correlation Spectroscopy: Criteria for Analysis in Complex Systems
  Alexei Tcherniak, Carmen Reznik, Stephan Link and Christy F. Landes
  Analytical Chemistry2009 81 (2), 746-754
  We have evaluated the effect of varying three key parameters for Fluorescence Correlation Spectroscopy analysis, first in the context of a one species/one environment system, and then in a complex system composed of two species, or conversely, two ...
  Abstract | Full Text HTML | Hi-Res PDF | PDF w/ Links
Molecular Fluorescence, Phosphorescence, and Chemiluminescence Spectrometry
Mark Lowry, Sayo O. Fakayode, Maxwell L. Geng, Gary A. Baker, Lin Wang, Matthew E. McCarroll, Gabor Patonay and Isiah M. Warner
Analytical Chemistry2008 80 (12), 4551-4574
- Molecular Fluorescence, Phosphorescence, and Chemiluminescence Spectrometry
  Mark Lowry, Sayo O. Fakayode, Maxwell L. Geng, Gary A. Baker, Lin Wang, Matthew E. McCarroll, Gabor Patonay and Isiah M. Warner
  Analytical Chemistry2008 80 (12), 4551-4574
  Abstract | Full Text HTML | Hi-Res PDF | PDF w/ Links