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Victor Hu, Martin J. Holland, Ian A. Cree, James Pullin, Helen A. Weiss, Patrick Massae, David C. Mabey, Robin L. Bailey, Matthew J. Burton, Phil Luthert; In Vivo Confocal Microscopy And Histopathology Of The Conjunctiva: A Systematic Comparison Involving Healthy And Trachomatous Conjunctiva. Invest. Ophthalmol. Vis. Sci. 2012;53(14):3991.
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In vivo confocal microscopy (IVCM) of the ocular surface allows high resolution images to be taken non-invasively and repeatedly. The interpretation of IVCM images, however, can be challenging. We have previously published IVCM grading protocols for inflammatory features and connective tissue scarring in the tarsal conjunctival surface. In order to validate the interpretation of images we conducted a study to systematically compare IVCM and immunohisotological analyses in healthy and diseased tissue.
Participants with healthy conjunctiva were recruited from patients undergoing cataract/retinal detachment surgery. Participants with diseased conjunctiva were recruited from patients with severe trachomatous scarring undergoing trichiasis surgery. IVCM of the tarsal conjunctival surface was performed using the HRT3/RCM and a conjunctival biopsy was taken which was fixed in formalin and embedded in paraffin. Histological sections were stained with hematoxylin and eosin and for CD45 and CD83. Masked, independent grading of IVCM images and biopsy sections was performed for: connective tissue scarring (using polarized light on histological sections to visualize collagen fibers); the inflammatory cell density (using H&E stained sections); and the presence of IVCM dendritifrom cells and immunohistochemical dendritic cells using (i) CD45+ cells with a dendritic morphology and (ii) CD83+ cells.
The histological and IVCM scarring grades showed good correlation (test for trend <0.001). A moderate association was found between histological and IVCM inflammatory cell density assessments (p=0.05). No significant association was found for the presence of IVCM dendritifrom cells and the presence of immunohistochemical dendritic cells.
The IVCM analysis of scarring in the tarsal conjunctiva is able to show good agreement with histological grading. The lack of association between the presence of IVCM dendritiform cells and immunohistochemical dendritic cells may be a result of study limitations. However, this discordance may be a result of the dendritiform structures representing something else, such as fibroblasts, which can also have marked dendritic extensions.
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