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Graphical representation and multicomponent analysis of single‐frequency fluorescence lifetime imaging microscopy data
Author(s) -
Clayton A. H. A.,
Hanley Q. S.,
Verveer P. J.
Publication year - 2004
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1111/j.1365-2818.2004.01265.x
Subject(s) - representation (politics) , weighting , microscopy , fluorescence , fluorescence microscope , biological system , fluorescence lifetime imaging microscopy , algorithm , exponential function , computer science , exponential growth , chemistry , optics , physics , mathematics , mathematical analysis , biology , acoustics , politics , political science , law
Summary Graphical representation of fluorescence lifetime imaging microscopy data demonstrates that a mixture of two components with single exponential decays can be resolved by single frequency measurements. We derive a method based on linear fitting that allows the calculation of the fluorescence lifetimes of the two components. We show that introduction of proper error‐weighting results in a non‐linear method that is mathematically identical to a global analysis algorithm that was recently derived. The graphical approach was applied to cellular data obtained from a lifetime‐based phosphorylation assay for the epidermal growth factor receptor and yielded results similar to those obtained by a global analysis algorithm.