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Journal of Biomedical Optics

Two-photon spectral fluorescence lifetime and second-harmonic generation imaging of the porcine cornea with a 12-femtosecond laser microscope
Author(s): Ana Batista; Hans Georg Breunig; Aisada Uchugonova; António Miguel Morgado; Karsten König
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Paper Abstract

Five dimensional microscopy with a 12-fs laser scanning microscope based on spectrally resolved two-photon autofluorescence lifetime and second-harmonic generation (SHG) imaging was used to characterize all layers of the porcine cornea. This setup allowed the simultaneous excitation of both metabolic cofactors, NAD(P)H and flavins, and their discrimination based on their spectral emission properties and fluorescence decay characteristics. Furthermore, the architecture of the stromal collagen fibrils was assessed by SHG imaging in both forward and backward directions. Information on the metabolic state and the tissue architecture of the porcine cornea were obtained with subcellular resolution, and high temporal and spectral resolutions.

Paper Details

Date Published: 8 March 2016
PDF: 11 pages
J. Biomed. Opt. 21(3) 036002 doi: 10.1117/1.JBO.21.3.036002
Published in: Journal of Biomedical Optics Volume 21, Issue 3
Show Author Affiliations
Ana Batista, Univ. des Saarlandes (Germany)
Univ. of Coimbra (Portugal)
Hans Georg Breunig, Univ. des Saarlandes (Germany)
JenLab GmbH (Germany)
Aisada Uchugonova, Univ. des Saarlandes (Germany)
António Miguel Morgado, Univ. de Coimbra (Portugal)
Karsten König, Saarland University (Germany)
JenLab GmbH (Germany)


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