Subcellular Comparison of Visible-Light Optical Coherence Tomography and Electron Microscopy in the Mouse Outer Retina
| Autor: | Pooja, Chauhan, Aaron M, Kho, Paul, FitzGerald, Bradley, Shibata, Vivek J, Srinivasan |
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| Rok vydání: | 2022 |
| Předmět: |
Microscopy
retina optical coherence tomography electron microscopy retinal pigment epithe-lium Neurosciences Bioengineering General Medicine Retinal Pigment Epithelium Biological Sciences Inbred C57BL Ophthalmology & Optometry Electron Medical and Health Sciences Retina Mice Inbred C57BL Microscopy Electron Mice inner segment Optical Coherence outer segment junction Animals Tomography Eye Disease and Disorders of Vision Tomography Optical Coherence |
| Zdroj: | Investigative ophthalmology & visual science, vol 63, iss 9 |
| Popis: | PurposeWe employed in vivo, 1.0-µm axial resolution visible-light optical coherence tomography (OCT) and ex vivo electron microscopy (EM) to investigate three subcellular features in the mouse outer retina: reflectivity oscillations inner to band 1 (study 1); hyperreflective band 2, attributed to the ellipsoid zone or inner segment/outer segment (IS/OS) junction (study 2); and the hyperreflective retinal pigment epithelium (RPE) within band 4 (study 3).MethodsPigmented (C57BL/6J, n = 10) and albino (BALB/cJ, n = 3) mice were imaged in vivo. Enucleated eyes were processed for light and electron microscopy. Using well-accepted reference surfaces, we compared micrometer-scale axial reflectivity of visible-light OCT with subcellular organization, as revealed by 9449 annotated EM organelles and features across four pigmented eyes.ResultsIn study 1, outer nuclear layer reflectivity peaks coincided with valleys in heterochromatin clump density (-0.34 ± 2.27 µm limits of agreement [LoA]). In study 2, band 2 depth on OCT and IS/OS junction depth on EM agreed (-0.57 ± 0.76 µm LoA), with both having similar distributions. In study 3, RPE electron dense organelle distribution did not agree with reflectivity in C57BL/6J mice, with OCT measures of RPE thickness exceeding those of EM (2.09 ± 0.89 µm LoA). Finally, RPE thickness increased with age in pigmented mice (slope = 0.056 µm/mo; P = 6.8 × 10-7).ConclusionsVisible-light OCT bands arise from subcellular organization, enabling new measurements in mice. Quantitative OCT-EM comparisons may be confounded by hydration level, particularly in the OS and RPE. Caution is warranted in generalizing results to other species. |
| Databáze: | OpenAIRE |
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