Applications of rapid time-gated hyperspectral FLIM: live cell imaging of membrane order and 6-D microscopy

H. B Manning; D. M Owen; E. Auksorius; P. A. A. de Beule; S. Oddos; C. B Talbot; C. Dunsby; I. Munro; A. I Magee; M. A. A. Neil; P. M. W. French

doi:10.1117/12.728479

13 July 2007 Applications of rapid time-gated hyperspectral FLIM: live cell imaging of membrane order and 6-D microscopy

H. B Manning, D. M Owen, E. Auksorius, P. A. A. de Beule, S. Oddos, C. B Talbot, C. Dunsby, I. Munro, A. I Magee, M. A. A. Neil, P. M. W. French

Author Affiliations +

Proceedings Volume 6630, Confocal, Multiphoton, and Nonlinear Microscopic Imaging III; 663019 (2007) https://doi.org/10.1117/12.728479
Event: European Conferences on Biomedical Optics, 2007, Munich, Germany

Abstract

We describe the characterisation of a hyperspectral fluorescence lifetime imaging microscope that exploits high-speed time-gated imaging technology and a tunable continuum source for 6-D fluorescence imaging. This line-scanning confocal microscope can record the full spectral-temporal (i.e. excitation-emission-lifetime) fluorescence matrix at each pixel in a three dimensional (x-y-z) sample. This instrument has been applied to biological samples including model membranes and live cells labelled with the phase-sensitive membrane dye di-4-ANEPPDHQ, for which significant variation of lifetime with emission wavelength is observed.

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H. B Manning, D. M Owen, E. Auksorius, P. A. A. de Beule, S. Oddos, C. B Talbot, C. Dunsby, I. Munro, A. I Magee, M. A. A. Neil, and P. M. W. French "Applications of rapid time-gated hyperspectral FLIM: live cell imaging of membrane order and 6-D microscopy", Proc. SPIE 6630, Confocal, Multiphoton, and Nonlinear Microscopic Imaging III, 663019 (13 July 2007); https://doi.org/10.1117/12.728479

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