Aims To describe the in vivo confocal microscopy (IVCM) features of human limbal nerve corpuscles (LNCs) and correlate these with the histological features.
Methods We examined 40 eyes of 29 healthy living subjects (17 female, 12 male; mean age=47.6) by IVCM. Four limbal quadrants were scanned through all epithelial layers and stroma to identify the LNCs and associated nerves. Ten fresh normal human corneoscleral discs from five deceased patients with a mean age of 67 years and 17 eye-bank corneoscleral rims with a mean age of 57.6 years were stained as whole mounts by the acetylcholinesterase (AChE) method to demonstrate LNCs and corneal nerves. Stained tissue was scanned in multiple layers with the NanoZoomer digital pathology microscope. The in vivo results were correlated to the histological findings.
Results On IVCM, LNCs were identified in 65% of the eyes studied and were mainly (84%) located in the inferior or superior limbal regions. They appeared either as bright (hyper-reflective) round or oval single structures within the hyporeflective, relatively acellular fibrous core of the palisades or were clustered in groups, often located anterior to the palisades of Vogt. They measured 36 µm in largest diameter (range 20–56 µm). The in vivo features were consistent with the histology, which showed LNCs as strongly AChE positive round or oval structures.
Conclusion The strong correlation with histology will enable use of IVCM to study LNCs in normal and disease conditions.
- Experimental laboratory
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Contributors MAA-A conceived of the study, examination of subjects with IVCM, data analysis and drafting the manuscript. FSA, IM and ADSY carried out the processing of the histological samples. WMA and IM contributed to data analysis, revision of the manuscript. HSSD contributed to design of the work; analysis and interpretation of data, drafting the work. All authors read and approved the final manuscript.
Funding This research is funded by Fight for Sight through small grant award.
Competing interests HSSD: Honoraria and Travel expenses from Dompe, Croma, Santen, Allergan, Thea, Shares in NuVision and Glaxosmithkline.
Patient consent for publication Not required.
Ethics approval The research was conducted at University of Nottingham, Nottingham University Hospital NHS Trust, Queens medical centre, UK and approved by the East Midlands Research Ethics Committee (REC no. 06/Q2403/46).
Provenance and peer review Not commissioned; externally peer reviewed.
Data availability statement All data relevant to the study are included in the article or uploaded as online supplementary information.