Article ; Online: Homo-FRET Based Biosensors and Their Application to Multiplexed Imaging of Signalling Events in Live Cells
International Journal of Molecular Sciences, Vol 16, Iss 7, Pp 14695-
2015 Volume 14716
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 ... ...
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. |
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Keywords | time resolved fluorescence anisotropy imaging (TR-FAIM) ; FRET ; multiplexed imaging ; homo-FRET ; Biology (General) ; QH301-705.5 ; Chemistry ; QD1-999 |
Language | English |
Publishing date | 2015-06-01T00:00:00Z |
Publisher | MDPI AG |
Document type | Article ; Online |
Database | BASE - Bielefeld Academic Search Engine (life sciences selection) |
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