Development of a hyperspectral fluorescence lifetime imaging microscope and its application to tissue imaging

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

doi:10.1117/12.700081

19 February 2007 Development of a hyperspectral fluorescence lifetime imaging microscope and its application to tissue imaging

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

Author Affiliations +

Proceedings Volume 6441, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues V; 64411K (2007) https://doi.org/10.1117/12.700081
Event: SPIE BiOS, 2007, San Jose, California, United States

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.

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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, and Paul M. W. French "Development of a hyperspectral fluorescence lifetime imaging microscope and its application to tissue imaging", Proc. SPIE 6441, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues V, 64411K (19 February 2007); https://doi.org/10.1117/12.700081

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