Premium
Improved spatial discrimination of protein reaction states in cells by global analysis and deconvolution of fluorescence lifetime imaging microscopy data
Author(s) -
Verveer P. J.,
Squire A.,
Bastiaens P. I. H.
Publication year - 2001
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 - 10.1046/j.1365-2818.2001.00917.x
Subject(s) - deconvolution , microscopy , fluorescence , fluorescence lifetime imaging microscopy , fluorescence microscope , pixel , image resolution , resolution (logic) , chemistry , optics , physics , computer science , artificial intelligence
The deconvolution of fluorescence lifetime imaging microscopy (FLIM) data that were processed with global analysis techniques is described. Global analysis of FLIM data enables the determination of relative numbers of molecules in different protein reaction states on a pixel‐by‐pixel basis in cells. The three‐dimensional fluorescence distributions of each protein state can then be calculated and deconvolved. High‐resolution maps of the relative concentrations of each state are then obtained from the deconvolved images. We applied these techniques to quantitatively image the phosphorylation state of ErbB1 receptors tagged with green fluorescent protein in MCF7 cells.