Homo-FRET Based Biosensors and Their Application to Multiplexed Imaging of Signalling Events in Live Cells.
File(s)ijms-16-14695-manuscript - accepted.pdf (1.06 MB)
Accepted version
Author(s)
Warren, SC
Margineanu, A
Katan, M
Dunsby, C
French, PM
Type
Journal Article
Abstract
Multiplexed imaging of Förster Resonance Energy Transfer (FRET)-based biosensors potentially presents a powerful approach to monitoring the spatio-temporal correlation of signalling pathways within a single live cell. Here, we discuss the potential of homo-FRET based biosensors to facilitate multiplexed imaging. We demonstrate that the homo-FRET between pleckstrin homology domains of Akt (Akt-PH) labelled with mCherry may be used to monitor 3'-phosphoinositide accumulation in live cells and show how global analysis of time resolved fluorescence anisotropy measurements can be used to quantify this accumulation. We further present multiplexed imaging readouts of calcium concentration, using fluorescence lifetime measurements of TN-L15-a CFP/YFP based hetero-FRET calcium biosensor-with 3'-phosphoinositide accumulation.
Date Issued
2015-06-30
Date Acceptance
2015-06-17
Citation
International Journal of Molecular Sciences, 2015, 16 (7), pp.14695-14716
ISSN
1661-6596
Start Page
14695
End Page
14716
Journal / Book Title
International Journal of Molecular Sciences
Volume
16
Issue
7
Copyright Statement
© 2015 by the authors; licensee MDPI, Basel, Switzerland. This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
License URL
Identifier
PII: ijms160714695
Subjects
FRET
homo-FRET
multiplexed imaging
time resolved fluorescence anisotropy imaging (TR-FAIM)
Publication Status
Published