Homo-FRET Based Biosensors and Their Application to Multiplexed Imaging of Signalling Events in Live Cells

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 multipl...

Descripción completa

Detalles Bibliográficos
Autores principales: Warren, Sean C., Margineanu, Anca, Katan, Matilda, Dunsby, Chris, French, Paul M. W.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2015
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4519867/
https://www.ncbi.nlm.nih.gov/pubmed/26133241
http://dx.doi.org/10.3390/ijms160714695
_version_ 1782383567100182528
author Warren, Sean C.
Margineanu, Anca
Katan, Matilda
Dunsby, Chris
French, Paul M. W.
author_facet Warren, Sean C.
Margineanu, Anca
Katan, Matilda
Dunsby, Chris
French, Paul M. W.
author_sort Warren, Sean C.
collection PubMed
description 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.
format Online
Article
Text
id pubmed-4519867
institution National Center for Biotechnology Information
language English
publishDate 2015
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-45198672015-08-03 Homo-FRET Based Biosensors and Their Application to Multiplexed Imaging of Signalling Events in Live Cells Warren, Sean C. Margineanu, Anca Katan, Matilda Dunsby, Chris French, Paul M. W. Int J Mol Sci Article 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. MDPI 2015-06-30 /pmc/articles/PMC4519867/ /pubmed/26133241 http://dx.doi.org/10.3390/ijms160714695 Text en © 2015 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Warren, Sean C.
Margineanu, Anca
Katan, Matilda
Dunsby, Chris
French, Paul M. W.
Homo-FRET Based Biosensors and Their Application to Multiplexed Imaging of Signalling Events in Live Cells
title Homo-FRET Based Biosensors and Their Application to Multiplexed Imaging of Signalling Events in Live Cells
title_full Homo-FRET Based Biosensors and Their Application to Multiplexed Imaging of Signalling Events in Live Cells
title_fullStr Homo-FRET Based Biosensors and Their Application to Multiplexed Imaging of Signalling Events in Live Cells
title_full_unstemmed Homo-FRET Based Biosensors and Their Application to Multiplexed Imaging of Signalling Events in Live Cells
title_short Homo-FRET Based Biosensors and Their Application to Multiplexed Imaging of Signalling Events in Live Cells
title_sort homo-fret based biosensors and their application to multiplexed imaging of signalling events in live cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4519867/
https://www.ncbi.nlm.nih.gov/pubmed/26133241
http://dx.doi.org/10.3390/ijms160714695
work_keys_str_mv AT warrenseanc homofretbasedbiosensorsandtheirapplicationtomultiplexedimagingofsignallingeventsinlivecells
AT margineanuanca homofretbasedbiosensorsandtheirapplicationtomultiplexedimagingofsignallingeventsinlivecells
AT katanmatilda homofretbasedbiosensorsandtheirapplicationtomultiplexedimagingofsignallingeventsinlivecells
AT dunsbychris homofretbasedbiosensorsandtheirapplicationtomultiplexedimagingofsignallingeventsinlivecells
AT frenchpaulmw homofretbasedbiosensorsandtheirapplicationtomultiplexedimagingofsignallingeventsinlivecells

Ejemplares similares