PURPOSE: To demonstrate the application of multiphoton fluorescence (MF) and second harmonic generation (SHG) microscopy for ex vivo characterization of the structural alterations of human corneas due to keratoconus. METHODS: Buttons of keratoconic corneas derived from penetrating keratoplasty were sent for structural analysis with a custom-built multiphoton microscope. Fluorescence detected within the cytoplasm and the SHG signal obtained from collagen were used to demonstrate the morphologic changes in the corneal specimens. RESULTS: The fluorescent epithelial cells around the apical area were elongated and were aligned parallel to the adjacent collagen fibers. Parallel and centripetal distribution patterns of stromal collagen bundles were demonstrated at different depths within the keratoconic corneas. CONCLUSIONS: MF and SHG microscopy provides three-dimensional structural analysis of keratoconus ex vivo. It may provide important morphologic information for the investigation of the pathogenesis of keratoconus and may have potential in a clinical setting as an in vivo diagnostic and monitoring system for advancing keratoconus.
PURPOSE: To demonstrate the application of multiphoton fluorescence (MF) and second harmonic generation (SHG) microscopy for ex vivo characterization of the structural alterations of human corneas due to keratoconus. METHODS: Buttons of keratoconic corneas derived from penetrating keratoplasty were sent for structural analysis with a custom-built multiphoton microscope. Fluorescence detected within the cytoplasm and the SHG signal obtained from collagen were used to demonstrate the morphologic changes in the corneal specimens. RESULTS: The fluorescent epithelial cells around the apical area were elongated and were aligned parallel to the adjacent collagen fibers. Parallel and centripetal distribution patterns of stromal collagen bundles were demonstrated at different depths within the keratoconic corneas. CONCLUSIONS: MF and SHG microscopy provides three-dimensional structural analysis of keratoconus ex vivo. It may provide important morphologic information for the investigation of the pathogenesis of keratoconus and may have potential in a clinical setting as an in vivo diagnostic and monitoring system for advancing keratoconus.
Authors: M Vielreicher; S Schürmann; R Detsch; M A Schmidt; A Buttgereit; A Boccaccini; O Friedrich Journal: J R Soc Interface Date: 2013-07-17 Impact factor: 4.118
Authors: Moritz Winkler; Golroxan Shoa; Yilu Xie; Steven J Petsche; Peter M Pinsky; Tibor Juhasz; Donald J Brown; James V Jester Journal: Invest Ophthalmol Vis Sci Date: 2013-11-05 Impact factor: 4.799
Authors: Jang Hyuk Lee; Jose J Rico-Jimenez; Chi Zhang; Aneesh Alex; Eric J Chaney; Ronit Barkalifa; Darold R Spillman; Marina Marjanovic; Zane Arp; Steve R Hood; Stephen A Boppart Journal: Biomed Opt Express Date: 2019-10-01 Impact factor: 3.732