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Proceedings Paper

Development of a time-gated fluorescence lifetime microscope for in vivo corneal metabolic imaging
Author(s): Susana F. Silva; Ana Batista; Olga Ciutad Castejón; Maria João Quadrado; José Paulo Domingues; Miguel Morgado
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Paper Abstract

Metabolic imaging can be a valuable tool in the early diagnosis of corneal diseases. Cell metabolic changes can be assessed through non-invasive optical methods due to the autofluorescence of metabolic co-factors nicotinamide adenine dinucleotide (NADH) and flavin adenine dinucleotide (FAD). Both molecules exhibit double exponential fluorescence decays, with well-separated short and long lifetime components, which are related to their protein-bound and free states. Corneal metabolism can be monitored by measuring the relative contribution of these two components.

Here we report on the development of a fluorescence lifetime imaging microscope for in vivo measurement of FAD fluorescence lifetimes in corneal cells. The microscope is based on one-photon fluorescence excitation, through a pulsed blue diode laser. Fluorescence lifetime imaging is achieved using the Time-Gated technique. Structured illumination is used to improve the low axial resolution of wide-field time-gated FLIM. A Digital Micromirror Device (DMD) is used to produce the sinusoidal patterns required by structural illumination. The DMD control is integrated with the acquisition software of the imaging system which is based on an ultra-high speed gated image intensifier coupled to a CCD camera.

We present preliminary results concerning optical and timing performance of the fluorescence lifetime microscope. Preliminary tests with ex-vivo bovine corneas are also described.

Paper Details

Date Published: 15 July 2015
PDF: 9 pages
Proc. SPIE 9537, Clinical and Biomedical Spectroscopy and Imaging IV, 953709 (15 July 2015); doi: 10.1117/12.2183945
Show Author Affiliations
Susana F. Silva, Univ. of Coimbra (Portugal)
Ana Batista, Univ. of Coimbra (Portugal)
Saarland Univ. (Germany)
Olga Ciutad Castejón, Polytechnical Univ. of Catalonia (Spain)
Maria João Quadrado, Univ. of Coimbra (Portugal)
Coimbra Hospital and Univ. Ctr. (Portugal)
José Paulo Domingues, Univ. of Coimbra (Portugal)
Miguel Morgado, Univ. of Coimbra (Portugal)

Published in SPIE Proceedings Vol. 9537:
Clinical and Biomedical Spectroscopy and Imaging IV
J. Quincy Brown; Volker Deckert, Editor(s)

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