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Dynamic imaging of cytosolic zinc in A rabidopsis roots combining FRET sensors and RootChip technology
Author(s) -
Lanquar Viviane,
Grossmann Guido,
Vinkenborg Jan L.,
Merkx Maarten,
Thomine Sébastien,
Frommer Wolf B.
Publication year - 2014
Publication title -
new phytologist
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.742
H-Index - 244
eISSN - 1469-8137
pISSN - 0028-646X
DOI - 10.1111/nph.12652
Subject(s) - förster resonance energy transfer , cytosol , biophysics , chemistry , zinc , microfluidics , live cell imaging , fluorescence , metal ions in aqueous solution , biochemistry , enzyme , nanotechnology , metal , cell , biology , materials science , physics , quantum mechanics , organic chemistry
Summary Zinc plays a central role in all living cells as a cofactor for enzymes and as a structural element enabling the adequate folding of proteins. In eukaryotic cells, metals are highly compartmentalized and chelated. Although essential to characterize the mechanisms of Zn 2+ homeostasis, the measurement of free metal concentrations in living cells has proved challenging and the dynamics are difficult to determine. Our work combines the use of genetically encoded Förster resonance energy transfer ( FRET ) sensors and a novel microfluidic technology, the RootChip, to monitor the dynamics of cytosolic Zn 2+ concentrations in A rabidopsis root cells. Our experiments provide estimates of cytosolic free Zn 2+ concentrations in A rabidopsis root cells grown under sufficient (0.4 nM) and excess (2 nM) Zn 2+ supply. In addition, monitoring the dynamics of cytosolic [ Zn 2+ ] in response to external supply suggests the involvement of high‐ and low‐affinity uptake systems as well as release from internal stores. In this study, we demonstrate that the combination of genetically encoded FRET sensors and microfluidics provides an attractive tool to monitor the dynamics of cellular metal ion concentrations over a wide concentration range in root cells.