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Combining ocFLIM and FIDSAM reveals fast and dynamic physiological responses at subcellular resolution in living plant cells
Author(s) -
ELGASS K.,
CAESAR K.,
HARTER K.,
MEIXNER A.J.,
SCHLEIFENBAUM F.
Publication year - 2011
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.1365-2818.2010.03446.x
Subject(s) - autofluorescence , subcellular localization , green fluorescent protein , fluorescence microscope , biophysics , microscopy , fluorescence , fluorescence lifetime imaging microscopy , intracellular , arabidopsis , fusion protein , protein subcellular localization prediction , fusion , biology , microbiology and biotechnology , biochemistry , physics , recombinant dna , optics , cytoplasm , linguistics , philosophy , gene , mutant
Summary For a deeper understanding of molecular mechanisms within cells and for the realization of predictive biology for intracellular processes at subcellular level, quantitative biology is required. Therefore, novel optical and spectroscopic technologies with quantitative and dynamic output are needed in cell biology. Here, we present a combined approach of novel one‐chromophore fluorescence lifetime imaging microscopy to probe the local environment of fluorescent fusion proteins and fluorescence intensity decay shape analysis microscopy to suppress interfering autofluorescence. By applying these techniques, we are able to analyse the subcellular localization and partitioning of a green fluorescence protein fusion of the salt stress‐induced protein low temperature induced (LTI)6b in great detail with high spatial and temporal resolution in living cells of Arabidopsis plants.