3D-Reconstruction of the human conventional outflow system by ribbon scanning confocal microscopy

The risk for glaucoma is driven by the microanatomy and function of the anterior segment. We performed a computation-intense, high-resolution, full-thickness ribbon-scanning confocal microscopy (RSCM) of the outflow tract of two human eyes. We hypothesized this would reveal important species differe...

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Bibliographic Details
Published in:PloS one 2020-05, Vol.15 (5), p.e0232833-e0232833
Main Authors: Loewen, Ralitsa T, Waxman, Susannah, Wang, Chao, Atta, Sarah, Chen, Si, Watkins, Simon C, Watson, Alan M, Loewen, Nils A
Format: Article
Language:English
Subjects:
Animals
Aqueous Humor - physiology
Benzoates
Benzyl alcohol
Biology and Life Sciences
Blood vessels
Cellular biology
Confocal microscopy
Development and progression
Diagnosis
Eye
Eye examination
Fluorescent Dyes
Glaucoma
High resolution
Human subjects
Humans
Laboratories
Lectins
Medicine and Health Sciences
Methods
Microscopy
Outflow
Plant Lectins
Reconstruction
Research and Analysis Methods
Rhodamines
Risk factors
Scanning
Sclera - blood supply
Sclera - ultrastructure
Species Specificity
Swine - anatomy & histology
Thickness
Trabecular Meshwork - ultrastructure
Transplants & implants
Veins - ultrastructure
Work stations
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Summary:The risk for glaucoma is driven by the microanatomy and function of the anterior segment. We performed a computation-intense, high-resolution, full-thickness ribbon-scanning confocal microscopy (RSCM) of the outflow tract of two human eyes. We hypothesized this would reveal important species differences when compared to existing data of porcine eyes, an animal that does not spontaneously develop glaucoma. After perfusing two human octogenarian eyes with lectin-fluorophore conjugate and optical clearance with benzyl alcohol benzyl benzoate (BABB), anterior segments were scanned by RSCM and reconstructed in 3D for whole-specimen rendering. Morphometric analyses of the outflow tract were performed for the trabecular meshwork (TM), limbal, and perilimbal outflow structures and compared to existing porcine data. RSCM provided high-resolution data for IMARIS-based surface reconstruction of outflow tract structures in 3D. Different from porcine eyes with an abundance of highly interconnected, narrow, and short collector channels (CCs), human eyes demonstrated fewer CCs which had a 1.5x greater cross-sectional area (CSA) and 2.6x greater length. Proximal CC openings at the level of Schlemm's canal (SC) had a 1.3x larger CSA than distal openings into the scleral vascular plexus (SVP). CCs were 10.2x smaller in volume than the receiving SVP vessels. Axenfeld loops, projections of the long ciliary nerve, were also visualized. In this high-resolution, volumetric RSCM analysis, human eyes had far fewer outflow tract vessels than porcine eyes. Human CCs spanned several clock-hours and were larger than in porcine eyes. These species differences may point to factors downstream of the TM that increase our vulnerability to glaucoma.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0232833