Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

    Full text documents are added when available. The database is updated daily and currently holds about 240,000 items, of which 72,000 in open access.

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Record number 419116
Title Wide-Field Multi-Parameter FLIM: Long-Term Minimal Invasive Observation of Proteins in Living Cells.
Author(s) Vitali, M.; Picazo, F.; Prokazov, Y.; Duci, A.; Turbin, E.; Götze, C.; Llopis, J.; Hartig, R.; Visser, A.J.W.G.; Zuschratter, W.
Source PLoS One 6 (2011)2. - ISSN 1932-6203 - 12 p.
DOI http://dx.doi.org/10.1371/journal.pone.0015820
Department(s) Biochemistry
Publication type Refereed Article in a scientific journal
Publication year 2011
Keyword(s) lifetime imaging microscopy - green fluorescent protein - resonance energy-transfer - single molecules - fret microscopy - global analysis - excitation - resolution - transport - oxygen
Abstract Time-domain Fluorescence Lifetime Imaging Microscopy (FLIM) is a remarkable tool to monitor the dynamics of fluorophore-tagged protein domains inside living cells. We propose a Wide-Field Multi-Parameter FLIM method (WFMP-FLIM) aimed to monitor continuously living cells under minimum light intensity at a given illumination energy dose. A powerful data analysis technique applied to the WFMP-FLIM data sets allows to optimize the estimation accuracy of physical parameters at very low fluorescence signal levels approaching the lower bound theoretical limit. We demonstrate the efficiency of WFMP-FLIM by presenting two independent and relevant long-term experiments in cell biology: 1) FRET analysis of simultaneously recorded donor and acceptor fluorescence in living HeLa cells and 2) tracking of mitochondrial transport combined with fluorescence lifetime analysis in neuronal processes.
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