Imperial College London

ProfessorDavidCarling

Faculty of MedicineInstitute of Clinical Sciences

Professor of Biochemistry
 
 
 
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Contact

 

+44 (0)20 3313 4313david.carling

 
 
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Location

 

3rd Fl CRBHammersmith HospitalHammersmith Campus

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Summary

 

Publications

Citation

BibTex format

@article{Chennell:2016:10.3390/s16081312,
author = {Chennell, G and Willows, RJW and Warren, SC and Carling, D and French, PMW and Dunsby, C and Sardini, A},
doi = {10.3390/s16081312},
journal = {Sensors},
title = {Imaging of Metabolic Status in 3D Cultures with an Improved AMPK FRET Biosensor for FLIM},
url = {http://dx.doi.org/10.3390/s16081312},
volume = {16},
year = {2016}
}

RIS format (EndNote, RefMan)

TY  - JOUR
AB - We describe an approach to non-invasively map spatiotemporal biochemical and physiological changes in 3D cell culture using Forster Resonance Energy Transfer (FRET) biosensors expressed in tumour spheroids. In particular, we present an improved Adenosine Monophosphate (AMP) Activated Protein Kinase (AMPK) FRET biosensor, mTurquoise2 AMPK Activity Reporter (T2AMPKAR), for fluorescence lifetime imaging (FLIM) readouts that we have evaluated in 2D and 3D cultures. Our results in 2D cell culture indicate that replacing the FRET donor, enhanced Cyan Fluorescent Protein (ECFP), in the original FRET biosensor, AMPK activity reporter (AMPKAR), with mTurquoise2 (mTq2FP), increases the dynamic range of the response to activation of AMPK, as demonstrated using the direct AMPK activator, 991. We demonstrated 3D FLIM of this T2AMPKAR FRET biosensor expressed in tumour spheroids using two-photon excitation.
AU - Chennell,G
AU - Willows,RJW
AU - Warren,SC
AU - Carling,D
AU - French,PMW
AU - Dunsby,C
AU - Sardini,A
DO - 10.3390/s16081312
PY - 2016///
SN - 1424-8239
TI - Imaging of Metabolic Status in 3D Cultures with an Improved AMPK FRET Biosensor for FLIM
T2 - Sensors
UR - http://dx.doi.org/10.3390/s16081312
UR - http://hdl.handle.net/10044/1/39026
VL - 16
ER -