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Proceedings Paper

Development of a hyperspectral fluorescence lifetime imaging microscope and its application to tissue imaging
Author(s): Dylan M. Owen; Hugh B. Manning; Pieter de Beule; Clifford Talbot; Jose Requejo-Isidro; Chris Dunsby; James McGinty; Richard K. P. Benninger; Dan S. Elson; Ian Munro; Neil P. Galletly; M. Jon Lever; Gordon W. Stamp; Praveen Anand; Mark A. A. Neil; Paul M. W. French
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Paper Abstract

We present the design, characterization and application of a novel, rapid, optically sectioned hyperspectral fluorescence lifetime imaging (FLIM) microscope. The system is based on a line scanning confocal configuration and uses a highspeed time-gated detector to extract lifetime information from many pixels in parallel. This allows the full spectraltemporal profiles of a fluorescence decay to be obtained from every pixel in an image. Line illumination and slit detection also gives the microscope a confocal optical sectioning ability. The system is applied to test samples and unstained biological tissue. In future, this microscope will be combined with recently-developed continuously electronically tunable, pulsed light sources based on tapered, micro-structured optical fibers. This will allow hyperspectral FLIM to be combined with the advantages of excitation spectroscopy to gain further insight into complex biological specimens including tissue and live cell imaging.

Paper Details

Date Published: 19 February 2007
PDF: 9 pages
Proc. SPIE 6441, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues V, 64411K (19 February 2007); doi: 10.1117/12.700081
Show Author Affiliations
Dylan M. Owen, Imperial College London (United Kingdom)
Hugh B. Manning, Imperial College London (United Kingdom)
Pieter de Beule, Imperial College London (United Kingdom)
Clifford Talbot, Imperial College London (United Kingdom)
Jose Requejo-Isidro, Imperial College London (United Kingdom)
Chris Dunsby, Imperial College London (United Kingdom)
James McGinty, Imperial College London (United Kingdom)
Richard K. P. Benninger, Imperial College London (United Kingdom)
Dan S. Elson, Imperial College London (United Kingdom)
Ian Munro, Imperial College London (United Kingdom)
Neil P. Galletly, Imperial College London (United Kingdom)
M. Jon Lever, Imperial College London (United Kingdom)
Gordon W. Stamp, Imperial College London (United Kingdom)
Praveen Anand, Imperial College London (United Kingdom)
Mark A. A. Neil, Imperial College London (United Kingdom)
Paul M. W. French, Imperial College London (United Kingdom)


Published in SPIE Proceedings Vol. 6441:
Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues V
Daniel L. Farkas; Robert C. Leif; Dan V. Nicolau, Editor(s)

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