Revisiting the Cornea and Trabecular Meshwork Junction With 2-Photon Excitation Fluorescence Microscopy
Autor: | Roy S. Chuck, Jason A. Liao, Catherine M. Marando, Choul Yong Park, Jimmy K. Lee |
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Rok vydání: | 2017 |
Předmět: |
0301 basic medicine
Materials science genetic structures Eye Banks Cornea Tendons 03 medical and health sciences 0302 clinical medicine Ciliary body Imaging Three-Dimensional Microscopy Electron Transmission Trabecular Meshwork Microscopy medicine Fluorescence microscope Humans biology Anatomy Fluorescence eye diseases Tissue Donors Elastin Ophthalmology 030104 developmental biology medicine.anatomical_structure Microscopy Fluorescence Multiphoton 030221 ophthalmology & optometry Biophysics biology.protein sense organs Trabecular meshwork Collagen Excitation |
Zdroj: | Cornea. 36(6) |
ISSN: | 1536-4798 |
Popis: | To investigate the collagen and elastin architecture at the junction of the human cornea and trabecular meshwork (TM).The cornea, TM, and ciliary body (CB) tendons of unfixed human corneal buttons were imaged with an inverted 2-photon excited fluorescence microscope (FluoView FV-1000; Olympus, Central Valley, PA). The laser (Ti:sapphire) was tuned to 850 nm for 2-photon excitation. Backscatter signals of second harmonic generation and autofluorescence were collected through a 425/30-nm emission filter and a 525/45-nm emission filter, respectively. The second harmonic generation signal corresponds to collagen fibers, and the autofluorescence signal corresponds to elastin-containing tissue. Tissue structure representations were obtained through software-generated reconstructions of consecutive and overlapping (z-stack) images through a relevant sample depth.Collagen-rich CB tendons insert into the cornea between Descemet membrane (DM) and posterior stroma along with elastin fibers originating from the TM. The CB tendons directly abut DM, and their insertion narrows as they course centrally in the cornea, giving a wedge appearance to these parallel collagen fibers. Approximately 260 μm centrally from the edge of DM, the CB tendons fan out and merge with pre-DM collagen. As the CB tendons enter the cornea, they form a dense collagenous comb-like structure orthogonal to the edge of DM and supported by a delicate elastin network of interwoven fibers originating from the TM.Two-photon excited fluorescence microscopy has improved our understanding of the peripheral corneal architecture. CB tendon insertions in this region may contribute to the radial tears encountered when preparing DM endothelial keratoplasty grafts. |
Databáze: | OpenAIRE |
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