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

International Journal of Molecular Sciences

Sean C. Warren²  Anca Margineanu²  Matilda Katan¹  Chris Dunsby²  Paul M. W. French² 

[1] Structural and Molecular Biology, University College London, London WC1E 6BT, UK; E-Mail:;Photonics Group, Department of Physics, Imperial College London, London SW7 2AZ, UK; E-Mails:

关键词: time resolved fluorescence anisotropy imaging (TR-FAIM);    FRET;    multiplexed imaging;    homo-FRET;   

DOI  :  10.3390/ijms160714695

来源: mdpi

【 摘 要 】

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.

【 授权许可】

CC BY   
© 2015 by the authors; licensee MDPI, Basel, Switzerland.