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22 February 2013 Enhanced resolution and sensitivity in fluorescence fluctuation measurements using multi-modal data acquisition and global analysis
Keith M. Berland, Neil R. Anthony
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Proceedings Volume 8588, Multiphoton Microscopy in the Biomedical Sciences XIII; 858801 (2013) https://doi.org/10.1117/12.2008872
Event: SPIE BiOS, 2013, San Francisco, California, United States
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.
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Keith M. Berland and Neil R. Anthony "Enhanced resolution and sensitivity in fluorescence fluctuation measurements using multi-modal data acquisition and global analysis", Proc. SPIE 8588, Multiphoton Microscopy in the Biomedical Sciences XIII, 858801 (22 February 2013); https://doi.org/10.1117/12.2008872
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KEYWORDS
Fluorescence correlation spectroscopy
Luminescence
Diffusion
Molecules
Data modeling
Data acquisition
Resolution enhancement technologies
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