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Novel Application of Fluorescence Lifetime and Fluorescence Microscopy Enables Quantitative Access to Subcellular Dynamics in Plant Cells

BACKGROUND: Optical and spectroscopic technologies working at subcellular resolution with quantitative output are required for a deeper understanding of molecular processes and mechanisms in living cells. Such technologies are prerequisite for the realisation of predictive biology at cellular and su...

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Autores principales: Elgass, Kirstin, Caesar, Katharina, Schleifenbaum, Frank, Stierhof, York-Dieter, Meixner, Alfred J., Harter, Klaus
Formato: Texto
Lenguaje:English
Publicado: Public Library of Science 2009
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2683565/
https://www.ncbi.nlm.nih.gov/pubmed/19492078
http://dx.doi.org/10.1371/journal.pone.0005716
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author Elgass, Kirstin
Caesar, Katharina
Schleifenbaum, Frank
Stierhof, York-Dieter
Meixner, Alfred J.
Harter, Klaus
author_facet Elgass, Kirstin
Caesar, Katharina
Schleifenbaum, Frank
Stierhof, York-Dieter
Meixner, Alfred J.
Harter, Klaus
author_sort Elgass, Kirstin
collection PubMed
description BACKGROUND: Optical and spectroscopic technologies working at subcellular resolution with quantitative output are required for a deeper understanding of molecular processes and mechanisms in living cells. Such technologies are prerequisite for the realisation of predictive biology at cellular and subcellular level. However, although established in the physical sciences, these techniques are rarely applied to cell biology in the plant sciences. PRINCIPAL FINDINGS: Here, we present a combined application of one-chromophore fluorescence lifetime microscopy and wavelength-selective fluorescence microscopy to analyse the function of a GFP fusion of the Brassinosteroid Insensitive 1 Receptor (BRI1-GFP) with high spatial and temporal resolution in living Arabidopsis cells in their tissue environment. We show a rapid, brassinolide-induced cell wall expansion and a fast BR-regulated change in the BRI1-GFP fluorescence lifetime in the plasmamembrane in vivo. Both cell wall expansion and changes in fluorescence lifetime reflect early BR-induced and BRI1-dependent physiological or signalling processes. Our experiments also show the potential of one-chromophore fluorescence lifetime microscopy for the in vivo monitoring of the biochemical and biophysical subcellular environment using GFP fusion proteins as probes. SIGNIFICANCE: One-chromophore fluorescence lifetime microscopy, combined with wavelength-specific fluorescence microscopy, opens up new frontiers for in vivo dynamic and quantitative analysis of cellular processes at high resolution which are not addressable by pure imaging technologies or transmission electron microscopy.
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spelling pubmed-26835652009-06-02 Novel Application of Fluorescence Lifetime and Fluorescence Microscopy Enables Quantitative Access to Subcellular Dynamics in Plant Cells Elgass, Kirstin Caesar, Katharina Schleifenbaum, Frank Stierhof, York-Dieter Meixner, Alfred J. Harter, Klaus PLoS One Research Article BACKGROUND: Optical and spectroscopic technologies working at subcellular resolution with quantitative output are required for a deeper understanding of molecular processes and mechanisms in living cells. Such technologies are prerequisite for the realisation of predictive biology at cellular and subcellular level. However, although established in the physical sciences, these techniques are rarely applied to cell biology in the plant sciences. PRINCIPAL FINDINGS: Here, we present a combined application of one-chromophore fluorescence lifetime microscopy and wavelength-selective fluorescence microscopy to analyse the function of a GFP fusion of the Brassinosteroid Insensitive 1 Receptor (BRI1-GFP) with high spatial and temporal resolution in living Arabidopsis cells in their tissue environment. We show a rapid, brassinolide-induced cell wall expansion and a fast BR-regulated change in the BRI1-GFP fluorescence lifetime in the plasmamembrane in vivo. Both cell wall expansion and changes in fluorescence lifetime reflect early BR-induced and BRI1-dependent physiological or signalling processes. Our experiments also show the potential of one-chromophore fluorescence lifetime microscopy for the in vivo monitoring of the biochemical and biophysical subcellular environment using GFP fusion proteins as probes. SIGNIFICANCE: One-chromophore fluorescence lifetime microscopy, combined with wavelength-specific fluorescence microscopy, opens up new frontiers for in vivo dynamic and quantitative analysis of cellular processes at high resolution which are not addressable by pure imaging technologies or transmission electron microscopy. Public Library of Science 2009-05-27 /pmc/articles/PMC2683565/ /pubmed/19492078 http://dx.doi.org/10.1371/journal.pone.0005716 Text en Elgass et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Elgass, Kirstin
Caesar, Katharina
Schleifenbaum, Frank
Stierhof, York-Dieter
Meixner, Alfred J.
Harter, Klaus
Novel Application of Fluorescence Lifetime and Fluorescence Microscopy Enables Quantitative Access to Subcellular Dynamics in Plant Cells
title Novel Application of Fluorescence Lifetime and Fluorescence Microscopy Enables Quantitative Access to Subcellular Dynamics in Plant Cells
title_full Novel Application of Fluorescence Lifetime and Fluorescence Microscopy Enables Quantitative Access to Subcellular Dynamics in Plant Cells
title_fullStr Novel Application of Fluorescence Lifetime and Fluorescence Microscopy Enables Quantitative Access to Subcellular Dynamics in Plant Cells
title_full_unstemmed Novel Application of Fluorescence Lifetime and Fluorescence Microscopy Enables Quantitative Access to Subcellular Dynamics in Plant Cells
title_short Novel Application of Fluorescence Lifetime and Fluorescence Microscopy Enables Quantitative Access to Subcellular Dynamics in Plant Cells
title_sort novel application of fluorescence lifetime and fluorescence microscopy enables quantitative access to subcellular dynamics in plant cells
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2683565/
https://www.ncbi.nlm.nih.gov/pubmed/19492078
http://dx.doi.org/10.1371/journal.pone.0005716
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