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Proceedings Paper

Toward two-photon excited fluorescence lifetime endomicroscopy (Conference Presentation)
Author(s): Charles-Henri Hage; Pierre Leclerc; Marc Fabert; Julien Brevier; Rémi Habert; Flavie Braud; Alexandre Kudlinski; Frédéric Louradour

Paper Abstract

Fluorescence lifetime imaging microscopy (FLIM) represents a powerful tool for biological studies. Endoscopic FLIM applied to the intracellular native biomarker NADH and FAD represents a promising mean for in vivo in situ malignant tissue diagnosis in the medical field. Else, 2-photon-excited fluorescence (2PEF) provides increased 3D resolution and imaging depth. But very few demonstrations about 2PEF lifetime measurement through a fiber have been reported and none about endoscopic 2P-FLIM through a practical fiber length (< 3m). Our group has recently demonstrated the possibility to efficiently deliver through a very long optical fiber the short and intense excitation pulses required for 2P-FLIM. Our goal is now to check that collecting fluorescence through the same endoscopic fiber does not deteriorate the lifetime measurement. Relying on the basis previously published in case of 1PEF by P. French and co-workers (J. Biophotonics, 2015), we have experimentally quantitatively evaluated the influence on the lifetime measurement of the fiber chromatic and intermodal dispersions. The main result is that the fiber contribution to the system impulse response function, even in the case of a 3-meter long double-clad optical fiber, does not hinder the separation between free and bound NADH states using FLIM. Related calibrations and measurements will be detailed. Ongoing experiments about the development of a 2P-FLIM endomicroscope on the basis of an previously reported 2P-endomicroscope (Ducourthial et al., Sc. Reports, 2015), used under various configurations (i.e. point measurement in the center of the 2P-endomicroscope image, averaged lifetime, binned endoscopic 2P-FLIM image), will be also presented.

Paper Details

Date Published: 24 April 2017
PDF: 1 pages
Proc. SPIE 10069, Multiphoton Microscopy in the Biomedical Sciences XVII, 100691H (24 April 2017); doi: 10.1117/12.2252638
Show Author Affiliations
Charles-Henri Hage, XLIM Institut de Recherche (France)
Pierre Leclerc, XLIM Institut de Recherche (France)
Marc Fabert, XLIM Institut de Recherche (France)
Julien Brevier, XLIM Institut de Recherche (France)
Rémi Habert, Lab. de Physics des Laser, Atomes, et Molécules, Univ. des Sciences et Technologies de Lille (France)
Flavie Braud, Lab. de Physics des Laser, Atomes, et Molécules, Univ. des Sciences et Technologies de Lille (France)
Alexandre Kudlinski, Lab. de Physics des Laser, Atomes, et Molécules, Univ. des Sciences et Technologies de Lille (France)
Frédéric Louradour, XLIM Institut de Recherche (France)

Published in SPIE Proceedings Vol. 10069:
Multiphoton Microscopy in the Biomedical Sciences XVII
Ammasi Periasamy; Peter T. C. So; Karsten König; Xiaoliang S. Xie, Editor(s)

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