Comparison of histologic findings in age-related macular degeneration with RPE flatmount images
Purpose: To visualize and analyze ex vivo flatmounted human RPE morphology from patients with age-related macular degeneration (AMD), and to compare the morphology with histologic findings. To establish whether the sub-RPE structures identified en face in RPE flatmount preparations are drusen with h...
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Molecular Vision
2019-02-01
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doaj-4cbdccc0ec98479fa7a920524745159a2020-11-25T00:01:20ZengMolecular VisionMolecular Vision1090-05351090-05352019-02-012517078Comparison of histologic findings in age-related macular degeneration with RPE flatmount imagesQing Zhang0Micah A. Chrenek1Shagun Bhatia2Alia Rashid3Salma Ferdous4Kevin J. Donaldson5Henry Skelton6Wenfei Wu7Thonnie Rose O. See8Yi Jiang9Nupur Dalal10John M. Nickerson11Hans E. Grossniklaus12Department of Ophthalmology, Emory University School of Medicine, Atlanta, Department of Ophthalmology, Emory University School of Medicine, Atlanta, Department of Ophthalmology, Emory University School of Medicine, Atlanta, Department of Ophthalmology, Emory University School of Medicine, Atlanta, Department of Ophthalmology, Emory University School of Medicine, Atlanta, Department of Ophthalmology, Emory University School of Medicine, Atlanta, Department of Ophthalmology, Emory University School of Medicine, Atlanta, Department of Ophthalmology, Emory University School of Medicine, Atlanta, Department of Ophthalmology, Emory University School of Medicine, Atlanta, Department of Mathematics and Statistics, Georgia State University, Atlanta, GADepartment of Ophthalmology, Emory University School of Medicine, Atlanta, Department of Ophthalmology, Emory University School of Medicine, Atlanta, Department of Pathology, Emory University School of Medicine, Atlanta, GAPurpose: To visualize and analyze ex vivo flatmounted human RPE morphology from patients with age-related macular degeneration (AMD), and to compare the morphology with histologic findings. To establish whether the sub-RPE structures identified en face in RPE flatmount preparations are drusen with histopathological registration in serial sections. To detect characteristic patterns found en face in RPE with the same structures in histological cross sections from eyes from cadavers of patients with AMD. Methods: Twenty-eight postmortem eyes from 14 patients (16 eyes with AMD and 12 age-matched control eyes) were oriented and microdissected yielding a RPE-choroid preparation. The tissues were flatmounted, stained with Alexa Fluor 635 Phalloidin (AF635-phalloidin) for f-actin and propidium iodide for DNA, and imaged using confocal microscopy. Portions of tissue from macular regions were processed for electron microscopic examination. After confocal imaging, the samples were remounted for histologic processing, embedded in paraffin, and serially sectioned perpendicular to the plane of the RPE-choroid sheet. Scaled two-dimensional (2D) maps of drusen locations found with the histological cross sections were constructed and correlated with the en face confocal microscopic images. Results: Twenty-eight postmortem eyes with a mean time of death to tissue preservation of 23.7 h (range 8.0–51 h) from 14 donors (seven women and seven men) with an average age of 78 years (range 60–93 years) were evaluated. Eight donors had AMD, and six served as controls. Scattered small, hard drusen were present in the periphery of the eyes with AMD and the healthy eyes. The macular region of the eyes with AMD contained small (<63 µm), medium (63.0–124 µm), and large ( ≥ 125 µm) drusen. The RPE was arranged in rosette-like structures overlying small drusen, attenuated overlying medium-sized drusen, and consisted of large multinucleated cells overlying large drusen. The RPE in the area of geographic atrophy was attenuated and depigmented. Conclusions: Confocal images of flatmounts from eyes with AMD showed RPE patterns overlying various types of drusen and geographic atrophy that correlated with histologic characteristics. We propose RPE repair mechanisms that may result in the patterns that we observed. http://www.molvis.org/molvis/v25/70/age-related macular degenerationretinal pigment epithelium |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Qing Zhang Micah A. Chrenek Shagun Bhatia Alia Rashid Salma Ferdous Kevin J. Donaldson Henry Skelton Wenfei Wu Thonnie Rose O. See Yi Jiang Nupur Dalal John M. Nickerson Hans E. Grossniklaus |
spellingShingle |
Qing Zhang Micah A. Chrenek Shagun Bhatia Alia Rashid Salma Ferdous Kevin J. Donaldson Henry Skelton Wenfei Wu Thonnie Rose O. See Yi Jiang Nupur Dalal John M. Nickerson Hans E. Grossniklaus Comparison of histologic findings in age-related macular degeneration with RPE flatmount images Molecular Vision age-related macular degeneration retinal pigment epithelium |
author_facet |
Qing Zhang Micah A. Chrenek Shagun Bhatia Alia Rashid Salma Ferdous Kevin J. Donaldson Henry Skelton Wenfei Wu Thonnie Rose O. See Yi Jiang Nupur Dalal John M. Nickerson Hans E. Grossniklaus |
author_sort |
Qing Zhang |
title |
Comparison of histologic findings in age-related macular degeneration with RPE flatmount images |
title_short |
Comparison of histologic findings in age-related macular degeneration with RPE flatmount images |
title_full |
Comparison of histologic findings in age-related macular degeneration with RPE flatmount images |
title_fullStr |
Comparison of histologic findings in age-related macular degeneration with RPE flatmount images |
title_full_unstemmed |
Comparison of histologic findings in age-related macular degeneration with RPE flatmount images |
title_sort |
comparison of histologic findings in age-related macular degeneration with rpe flatmount images |
publisher |
Molecular Vision |
series |
Molecular Vision |
issn |
1090-0535 1090-0535 |
publishDate |
2019-02-01 |
description |
Purpose: To visualize and analyze ex vivo flatmounted human RPE morphology from patients with age-related macular degeneration (AMD), and to compare the morphology with histologic findings. To establish whether the sub-RPE structures identified en face in RPE flatmount preparations are drusen with histopathological registration in serial sections. To detect characteristic patterns found en face in RPE with the same structures in histological cross sections from eyes from cadavers of patients with AMD.
Methods: Twenty-eight postmortem eyes from 14 patients (16 eyes with AMD and 12 age-matched control eyes) were oriented and microdissected yielding a RPE-choroid preparation. The tissues were flatmounted, stained with Alexa Fluor 635 Phalloidin (AF635-phalloidin) for f-actin and propidium iodide for DNA, and imaged using confocal microscopy. Portions of tissue from macular regions were processed for electron microscopic examination. After confocal imaging, the samples were remounted for histologic processing, embedded in paraffin, and serially sectioned perpendicular to the plane of the RPE-choroid sheet. Scaled two-dimensional (2D) maps of drusen locations found with the histological cross sections were constructed and correlated with the en face confocal microscopic images.
Results: Twenty-eight postmortem eyes with a mean time of death to tissue preservation of 23.7 h (range 8.0–51 h) from 14 donors (seven women and seven men) with an average age of 78 years (range 60–93 years) were evaluated. Eight donors had AMD, and six served as controls. Scattered small, hard drusen were present in the periphery of the eyes with AMD and the healthy eyes. The macular region of the eyes with AMD contained small (<63 µm), medium (63.0–124 µm), and large ( ≥
125 µm) drusen. The RPE was arranged in rosette-like structures overlying small drusen, attenuated overlying medium-sized drusen, and consisted of large multinucleated cells overlying large drusen. The RPE in the area of geographic atrophy was attenuated and depigmented.
Conclusions: Confocal images of flatmounts from eyes with AMD showed RPE patterns overlying various types of drusen and geographic atrophy that correlated with histologic characteristics. We propose RPE repair mechanisms that may result in the patterns that we observed. |
topic |
age-related macular degeneration retinal pigment epithelium |
url |
http://www.molvis.org/molvis/v25/70/ |
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