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Visualization of Cu 2+ uptake and release in plant cells by fluorescence lifetime imaging microscopy
Author(s) -
Hötzer Benjamin,
Ivanov Rumen,
Brumbarova Tzvetina,
Bauer Petra,
Jung Gregor
Publication year - 2012
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/j.1742-4658.2011.08434.x
Subject(s) - fluorescence , green fluorescent protein , fluorescence microscope , biophysics , fluorescence lifetime imaging microscopy , microscopy , chemistry , eukaryote , divalent , in vivo , plant cell , live cell imaging , bimolecular fluorescence complementation , biology , biochemistry , cell , gene , genome , physics , microbiology and biotechnology , organic chemistry , optics , quantum mechanics
A principal objective in life sciences is the visualization of biochemical processes. Fluorescence‐based techniques are widely used to demonstrate transport of relevant substances across cellular membranes. In this paper we report a novel noninvasive, real‐time fluorescence lifetime imaging microscopy method for visualizing uptake and release of divalent copper ions (Cu 2+ ) in vivo . For this purpose, we employed a green fluorescent protein (GFP) form able to change its fluorescence lifetime upon Cu 2+ binding. We demonstrate that this technique is selective for Cu 2+ . We show the reversible decrease of the fluorescence lifetime of GFP from 2.2 to 1.6 ns in Escherichia coli and from 1.8 to 1.3 ns in root cells of Arabidopsis after the addition of Cu 2+ . Cu 2+ uptake of epidermal tobacco cells leads to a drop of the GFP lifetime from 2.5 to 2.2 ns. In summary, the spatially resolved visualization of Cu 2+ distribution in vivo is demonstrated in prokaryote and eukaryote cells.