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Enhanced resolution and sensitivity in fluorescence fluctuation measurements using multi-modal data acquisition and global analysis
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Paper Abstract

Fluorescence correlation spectroscopy (FCS) and related fluctuation spectroscopy and microscopy methods have become important research tools that enable detailed investigations of the chemical and physical properties of molecules and molecular systems in a variety of complex environments. When analyzed successfully fluctuation measurements often provide unique information that is otherwise difficult to measure, such as molecular concentrations and interaction stoichiometry. However, information recovery via curve fitting of fluctuation data can present challenges due to limited resolution and/or problems with fitting model verification. We discuss a new approach to fluctuation data analysis coupling multi-modal fluorescence measurements and global analysis, and demonstrate how this approach can provide enhanced sensitivity and resolution in fluctuation measurements. We illustrate the approach using a combination of FCS and fluorescence lifetime measurements, here called τFCS, and demonstrate the capability to recover the concentration of two independent molecular species in a two component mixture even when the species have identical diffusion coefficients and molecular brightness values. This work was partially supported by NSF grants MCB0817966 and DMR0907435.

Paper Details

Date Published: 22 February 2013
PDF: 10 pages
Proc. SPIE 8588, Multiphoton Microscopy in the Biomedical Sciences XIII, 858801 (22 February 2013); doi: 10.1117/12.2008872
Show Author Affiliations
Keith M. Berland, Emory Univ. (United States)
Neil R. Anthony, Emory Univ. (United States)

Published in SPIE Proceedings Vol. 8588:
Multiphoton Microscopy in the Biomedical Sciences XIII
Ammasi Periasamy; Karsten König; Peter T. C. So, Editor(s)

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