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Endogenous two- and three-photon fluorescence of a biological substance with a picosecond ultrawide band laser source
Author(s): Claire Lefort; Laetitia Magnol; Philippe Lévêque; Véronique Blanquet
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Paper Abstract

Multiphoton microscopy (MPM) is a method for characterizing biological samples, becoming more and more established in life sciences labs thanks to its label-free imaging ability. In MPM, few biological substances have been highlighted as endogenously fluorescent, such as elastin, myosin, keratin, redox indicators, collagen or amino acids. Tryptophan, an amino acid fundamental in the synthesis of proteins, is known for its endogenous fluorescence. In this article, we propose to show an original solution specifically dedicated to multiphoton microscopy with an ultrawide band laser system. The specificity stands into the filtering system based on a prisms-line allowing spatial shaping of the spectrum. Our custom-designed multiphoton microscope, coupled with a picosecond ultrawide band laser correctly filtered spectrally, is adapted for charactering the two- and three-photon absorption ranges and imaging of tryptophan. This highlights in one hand that the use of a picosecond ultrawide band laser spectrally filtered does allow to reach both the two- and three-photon abortion (TPA and ThPA) ranges of this substance. In another hand, a quantitative comparison of the resulting images shows high differences in the image quality where the three-photon image looks better contrasted and better resolved than the two-photon one. An explanation of this highly interesting phenomena can be proposed with a study of the probability of presence of multiphoton processes involved and the cross section values of ThPA and TPA. This initiating work, cumulating an innovative multiphoton setup and interesting results, plays a crucial role for extending the label-free imaging ability of MPM.

Paper Details

Date Published: 24 May 2018
PDF: 7 pages
Proc. SPIE 10679, Optics, Photonics, and Digital Technologies for Imaging Applications V, 106791N (24 May 2018); doi: 10.1117/12.2306681
Show Author Affiliations
Claire Lefort, Institut de recherché XLIM, CNRS Univ. de Limoges (France)
Laetitia Magnol, Lab. PEIRENE, Univ. de Limoges (France)
Philippe Lévêque, Institut de recherché XLIM, CNRS Univ. de Limoges (France)
Véronique Blanquet, Lab. PEIRENE, Univ. de Limoges (France)


Published in SPIE Proceedings Vol. 10679:
Optics, Photonics, and Digital Technologies for Imaging Applications V
Peter Schelkens; Touradj Ebrahimi; Gabriel Cristóbal, Editor(s)

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