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A high speed multifocal multiphoton fluorescence lifetime imaging microscope for live-cell FRET imaging.
Poland, Simon P; Krstajic, Nikola; Monypenny, James; Coelho, Simao; Tyndall, David; Walker, Richard J; Devauges, Viviane; Richardson, Justin; Dutton, Neale; Barber, Paul; Li, David Day-Uei; Suhling, Klaus; Ng, Tony; Henderson, Robert K; Ameer-Beg, Simon M.
Afiliación
  • Poland SP; Division of Cancer Studies, Guy's Campus, Kings College, London, UK ; Randall Division of Cell and Molecular Biophysics, Guy's Campus, Kings College, London, UK.
  • Krstajic N; Institute for Integrated Micro and Nano Systems, School of Engineering, University of Edinburgh, Edinburgh, UK.
  • Monypenny J; Division of Cancer Studies, Guy's Campus, Kings College, London, UK ; Randall Division of Cell and Molecular Biophysics, Guy's Campus, Kings College, London, UK.
  • Coelho S; Division of Cancer Studies, Guy's Campus, Kings College, London, UK ; Randall Division of Cell and Molecular Biophysics, Guy's Campus, Kings College, London, UK.
  • Tyndall D; Institute for Integrated Micro and Nano Systems, School of Engineering, University of Edinburgh, Edinburgh, UK.
  • Walker RJ; Institute for Integrated Micro and Nano Systems, School of Engineering, University of Edinburgh, Edinburgh, UK ; Photon-Force Ltd., Edinburgh, UK.
  • Devauges V; Division of Cancer Studies, Guy's Campus, Kings College, London, UK ; Randall Division of Cell and Molecular Biophysics, Guy's Campus, Kings College, London, UK.
  • Richardson J; Institute for Integrated Micro and Nano Systems, School of Engineering, University of Edinburgh, Edinburgh, UK ; Photon-Force Ltd., Edinburgh, UK.
  • Dutton N; Institute for Integrated Micro and Nano Systems, School of Engineering, University of Edinburgh, Edinburgh, UK.
  • Barber P; Gray Institute for Radiation Oncology & Biology, University of Oxford, Old Road Campus Research Building, Roosevelt Drive, Oxford OX3 7DQ UK.
  • Li DD; Strathclyde Institute of Pharmacy and Biomedical Sciences, 161 Cathedral Street, Glasgow, G4 0RE, UK.
  • Suhling K; Department of Physics, King's College London, Strand, London, UK.
  • Ng T; Division of Cancer Studies, Guy's Campus, Kings College, London, UK ; Randall Division of Cell and Molecular Biophysics, Guy's Campus, Kings College, London, UK ; UCL Cancer Institute, Paul O'Gorman Building, University College London, London WC1E 6DD, UK.
  • Henderson RK; Institute for Integrated Micro and Nano Systems, School of Engineering, University of Edinburgh, Edinburgh, UK.
  • Ameer-Beg SM; Division of Cancer Studies, Guy's Campus, Kings College, London, UK ; Randall Division of Cell and Molecular Biophysics, Guy's Campus, Kings College, London, UK.
Biomed Opt Express ; 6(2): 277-96, 2015 Feb 01.
Article en En | MEDLINE | ID: mdl-25780724
We demonstrate diffraction limited multiphoton imaging in a massively parallel, fully addressable time-resolved multi-beam multiphoton microscope capable of producing fluorescence lifetime images with sub-50ps temporal resolution. This imaging platform offers a significant improvement in acquisition speed over single-beam laser scanning FLIM by a factor of 64 without compromising in either the temporal or spatial resolutions of the system. We demonstrate FLIM acquisition at 500 ms with live cells expressing green fluorescent protein. The applicability of the technique to imaging protein-protein interactions in live cells is exemplified by observation of time-dependent FRET between the epidermal growth factor receptor (EGFR) and the adapter protein Grb2 following stimulation with the receptor ligand. Furthermore, ligand-dependent association of HER2-HER3 receptor tyrosine kinases was observed on a similar timescale and involved the internalisation and accumulation or receptor heterodimers within endosomes. These data demonstrate the broad applicability of this novel FLIM technique to the spatio-temporal dynamics of protein-protein interaction.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: Biomed Opt Express Año: 2015 Tipo del documento: Article Pais de publicación: Estados Unidos
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: Biomed Opt Express Año: 2015 Tipo del documento: Article Pais de publicación: Estados Unidos