Document Detail

Cumulant analysis in fluorescence fluctuation spectroscopy.
MedLine Citation:
PMID:  15189894     Owner:  NLM     Status:  MEDLINE    
A novel technique for the analysis of fluorescence fluctuation experiments is introduced. Fluorescence cumulant analysis (FCA) exploits the factorial cumulants of the photon counts and resolves heterogeneous samples based on differences in brightness. A simple analytical model connects the cumulants of the photon counts with the brightness epsilon and the number of molecules N in the optical observation volume for each fluorescent species. To provide the tools for a rigorous error analysis of FCA, expressions for the variance of factorial cumulants are developed and tested. We compare theory with experiment by analyzing dye mixtures and simple fluorophore solutions with FCA. A comparison of FCA with photon-counting histogram (PCH) analysis, a related technique, shows that both methods give identical results within experimental uncertainty. Both FCA and PCH are restricted to data sampling times that are short compared to the diffusion time of molecules through the observation volume of the instrument. But FCA theory, in contrast to PCH, can be extended to treat arbitrary sampling times. Here, we derive analytical expressions for the second factorial cumulant as a function of the sampling time and demonstrate that the theory successfully models fluorescence fluctuation data.
Joachim D Müller
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, Non-P.H.S.; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Biophysical journal     Volume:  86     ISSN:  0006-3495     ISO Abbreviation:  Biophys. J.     Publication Date:  2004 Jun 
Date Detail:
Created Date:  2004-06-10     Completed Date:  2005-01-24     Revised Date:  2013-06-09    
Medline Journal Info:
Nlm Unique ID:  0370626     Medline TA:  Biophys J     Country:  United States    
Other Details:
Languages:  eng     Pagination:  3981-92     Citation Subset:  IM    
School of Physics and Astronomy, University of Minnesota, Minneapolis, Minnesota 55455, USA.
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MeSH Terms
Fluorescent Dyes / chemistry*
Models, Theoretical*
Spectrometry, Fluorescence / methods*
Grant Support
Reg. No./Substance:
0/Fluorescent Dyes

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