Journal article
Calibration of Probe Volume in Fluorescence Correlation Spectroscopy
Applied spectroscopy, Vol.61(9), pp.956-962
09/2007
DOI: 10.1366/000370207781745883
PMID: 17910792
Abstract
In fluorescence correlation spectroscopy (FCS), an accurate evaluation of the probe volume is the basis of correct interpretation of experimental data and solution of an appropriate diffusion model. Poor fitting convergence has been a problem in the determination of the dimensional parameters, the beam radius, omega, and the distance along the optical axis of the probe volume, l. In this work, the instability of fitting during the calibration process is investigated by examining the chi(2) surfaces. We demonstrate that the minimum of chi(2) in the omega dimension is well defined for both converging and diverging data. The difficulty of fitting comes from the l dimension. The uncertainty in l could be significantly larger than that in omega, as determined by F-statistics. A modified calibration process is recommended based on examining two data treatment methods, combining several short data sets into a single long run and averaging the correlation functions of several short data sets. It is found that by using the mean of several converging correlation functions from short data sets instead of a long time correlation, more stable and consistent dimensional parameters are extracted to define the probe volume.
Details
- Title: Subtitle
- Calibration of Probe Volume in Fluorescence Correlation Spectroscopy
- Creators
- Yi Gao - Department of Chemistry, the Optical Science and Technology Center, the Nanoscience and Nanotechnology Institute, and the Center for Biocatalysis and Bioprocessing, University of Iowa, Iowa City, Iowa, 52242Zhenming Zhong - Department of Chemistry, the Optical Science and Technology Center, the Nanoscience and Nanotechnology Institute, and the Center for Biocatalysis and Bioprocessing, University of Iowa, Iowa City, Iowa, 52242M. Lei Geng - Department of Chemistry, the Optical Science and Technology Center, the Nanoscience and Nanotechnology Institute, and the Center for Biocatalysis and Bioprocessing, University of Iowa, Iowa City, Iowa, 52242
- Resource Type
- Journal article
- Publication Details
- Applied spectroscopy, Vol.61(9), pp.956-962
- DOI
- 10.1366/000370207781745883
- PMID
- 17910792
- ISSN
- 0003-7028
- eISSN
- 1943-3530
- Language
- English
- Date published
- 09/2007
- Academic Unit
- Chemistry
- Record Identifier
- 9984216603902771
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