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Proceedings Paper

The fluorescence lifetime of BRI1-GFP as probe for the noninvasive determination of the membrane potential in living cells
Author(s): K. Elgass; K. Caesar; F. Schleifenbaum; A. J. Meixner; K. Harter
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Paper Abstract

As the excited state lifetime of a fluorescent molecule depends on its environment, it is possible to use it as a probe for physico-chemical parameters of the surrounding medium. Whereas this is well known for many solid guest/host systems, only few reports of quantitative, temporal resolved in vivo studies to monitor the nano-environment for a protein-coupled chromophore such as GFP are known from literature. Here we present a novel approach to determine the membrane potential of living (plant) cells based on the fluorescence lifetime (FLT) analysis of membrane-located GFP. By using confocal sample scanning microscopy (CSSM) combined with fluorescence lifetime imaging microscopy, we recently showed that the phytohormone brassinolide (BL) induces cell wall expansion and a decrease in the FLT of the BRI1-GFP in living cells of Arabidopsis thaliana seedlings. BRI1 is the dominant functional receptor for BL in Arabidopsis and locates to the plasma membrane. Although the dependence of the FLT of GFP on its physico-chemical environment such as pH-value, refractive index and pressure has been reported, the observed FLT decrease of BRI1-GFP in response to BL application could not be explained by these parameters. However, our in vivo FLT and CSSM analyses indicate that the BLinduced change in the FLT of BRI1-GFP is caused by hyperpolarisation of the plasma membrane (Em). Thus, our results indicate that BRI1-GFP serves as sensitive and non-invasive probe for recording the Em of the plasma membrane in living plant cells with high spatio-temporal resolution.

Paper Details

Date Published: 11 February 2010
PDF: 8 pages
Proc. SPIE 7568, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues VIII, 756804 (11 February 2010); doi: 10.1117/12.841646
Show Author Affiliations
K. Elgass, Eberhard Karls Univ. Tübingen (Germany)
K. Caesar, Eberhard Karls Univ. Tübingen (Germany)
F. Schleifenbaum, Eberhard Karls Univ. Tübingen (Germany)
A. J. Meixner, Eberhard Karls Univ. Tübingen (Germany)
K. Harter, Eberhard Karls Univ. Tübingen (Germany)

Published in SPIE Proceedings Vol. 7568:
Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues VIII
Daniel L. Farkas; Dan V. Nicolau; Robert C. Leif, Editor(s)

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