Forde, TS and Hanley, QS ORCID: https://orcid.org/0000-0002-8189-9550, 2005. Following FRET through five energy transfer steps: spectroscopic photobleaching, recovery of spectra, and a sequential mechanism of FRET. Photochemical & Photobiological Sciences, 4 (8), pp. 609-616. ISSN 1474-905X
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Abstract
We report the acquisition and analysis of spectrally resolved photobleaching data from a model system designed to exhibit FRET. Spectrally resolved photobleaching can be used to determine the presence of FRET in these systems and to investigate multi-step mechanisms of energy transfer. The model system was a previously described set of fluorescent beads consisting of a system of six fluorophores. In standard photobleaching experiments to determine FRET, bleaching of an acceptor molecule resulting in recovery of donor intensity or changes in photobleaching kinetics are used as indicators of FRET. Here, we use the Bateman equations to model growth and decay in a photobleaching experiment. Linked donor–acceptor growth and decay is used as an indicator of FRET. The apparatus required is relatively simple when compared to lifetime imaging systems. Several data analysis strategies, rigorous model building, global fitting procedures, and error analysis are presented. Using these procedures a five-step sequential mechanism of energy transfer was selected for these beads.
Item Type: | Journal article |
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Publication Title: | Photochemical & Photobiological Sciences |
Creators: | Forde, T.S. and Hanley, Q.S. |
Publisher: | Royal Society of Chemistry |
Place of Publication: | Cambridge |
Date: | 2005 |
Volume: | 4 |
Number: | 8 |
ISSN: | 1474-905X |
Identifiers: | Number Type 10.1039/b416478d DOI |
Rights: | © Royal Society of Chemistry |
Divisions: | Schools > School of Science and Technology |
Record created by: | EPrints Services |
Date Added: | 09 Oct 2015 10:29 |
Last Modified: | 09 Jun 2017 13:31 |
URI: | https://irep.ntu.ac.uk/id/eprint/13750 |
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