title: Two-photon spectral fluorescence lifetime and second-harmonic generation imaging of the porcine cornea with a 12-femtosecond laser microscope.
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.
authors.affiliation: Saarland University, Faculty of Physics and Mechatronics, Department of Biophotonics and Laser Technology, Campus A5.1, Saarbrücken 66123, GermanybUniversity of Coimbra, Faculty of Medicine, Institute for Biomedical Imaging and Life Sciences, Azinhaga San.
authors.forename: Ana
authors.initials: A
authors.lastname: Batista
authors.affiliation: Saarland University, Faculty of Physics and Mechatronics, Department of Biophotonics and Laser Technology, Campus A5.1, Saarbrücken 66123, GermanycJenLab GmbH, Schillerstr. 1, Jena 07745, Germany.
authors.forename: Hans Georg
authors.initials: HG
authors.lastname: Breunig
authors.affiliation: Saarland University, Faculty of Physics and Mechatronics, Department of Biophotonics and Laser Technology, Campus A5.1, Saarbrücken 66123, Germany.
authors.forename: Aisada
authors.initials: A
authors.lastname: Uchugonova
authors.affiliation: University of Coimbra, Faculty of Medicine, Institute for Biomedical Imaging and Life Sciences, Azinhaga Santa Comba-Celas, Coimbra 3000-548, PortugaldUniversity of Coimbra, Faculty of Sciences and Technology, Department of Physics, Rua Larga, Coimbra 300.
authors.forename: António Miguel
authors.initials: AM
authors.lastname: Morgado
authors.affiliation: Saarland University, Faculty of Physics and Mechatronics, Department of Biophotonics and Laser Technology, Campus A5.1, Saarbrücken 66123, GermanycJenLab GmbH, Schillerstr. 1, Jena 07745, Germany.
authors.forename: Karsten
authors.initials: K
authors.lastname: König
mesh_terms: D000818:Animals; D003315:Cornea / Q000000981:diagnostic imaging*; D004867:Equipment Design; D018613:Microscopy, Confocal / Q000379:methods*; D061848:Optical Imaging / Q000379:methods*; D013552:Swine