Document Detail


Correction for contaminant fluorescence in frequency-domain fluorometry.
MedLine Citation:
PMID:  3578773     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
We describe a general method to correct for contaminant fluorescence when using the technique of frequency-domain fluorometry. The method can be applied regardless of the origin of the background signal, from scattered light, impurity fluorescence, or both. The procedure requires measurement of the frequency-dependent phase and modulation of the background at enough frequencies to approximate the decay law of the background. We also describe a general method to propagate the uncertainties in the measured phase and modulation values into the corrected values. This propagation is necessary to ensure proper weighting of the frequency-dependent data in the least-squares fitting algorithms. The practical usefulness of this correction method is demonstrated using frequency-domain data for one and two component mixtures which were deliberately contaminated with scattered light and/or other fluorophores.
Authors:
J R Lakowicz; R Jayaweera; N Joshi; I Gryczynski
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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:  Analytical biochemistry     Volume:  160     ISSN:  0003-2697     ISO Abbreviation:  Anal. Biochem.     Publication Date:  1987 Feb 
Date Detail:
Created Date:  1987-05-22     Completed Date:  1987-05-22     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0370535     Medline TA:  Anal Biochem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  471-9     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Anthracenes
Fluorescence
Fluorescent Dyes
Light
Models, Theoretical
Oxazoles
Scattering, Radiation
Spectrometry, Fluorescence / methods*
Grant Support
ID/Acronym/Agency:
GM-29318/GM/NIGMS NIH HHS; GM-35154/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/Anthracenes; 0/Fluorescent Dyes; 0/Oxazoles; 1499-10-1/9,10-diphenylanthracene; 1806-34-4/POPOP

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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