A method for quantifying limbal stem cell niches using OCT imaging

Abstract

AimsTo evaluate the efficacy of Fourier domain-optical coherence tomography (FD-OCT) in imaging and quantifying the limbal palisades of Vogt and to correlate these images with histological findings.MethodsThe superior and inferior limbal region of both eyes of 50 healthy volunteers were imaged by FD-OCT. Images were processed and analysed using Matlab software. In vitro immunofluorescent staining of a cadaveric donor limbus was analysed to correlate the presence of stem cells in the visualised structures.ResultsFD-OCT could successfully visualise limbal crypts and the palisades of Vogt in the limbus region. Fluorescent labelling confirmed the presence of stem cells in these structures. The mean palisade ridge width (ΔPR) and the mean interpalisade epithelial rete peg width (ΔERP) were both of the order of 72 μm, leading to a palisade density (PD) of about 7.4 palisades/mm. A significant difference in ΔPR, ΔERP and PD was seen between the inferior and superior sides of the right eye and the superior sides of the left and right eye(p<0.05.). A significant influence of iris colour on parameters ΔPR, ΔERP and PD was found, and of age on PD and ΔERP (p<0.05).ConclusionsIn vivo OCT imaging is a safe and effective modality to image the limbus and can be used to visualise the palisades of Vogt. Image processing using Matlab software enabled quantification and density calculation of imaged limbal palisades of Vogt. This technique may enhance targeted limbal biopsies for transplantation.

In Vivo Evaluation of the Limbus Using Anterior Segment Optical Coherence Tomography

“…Fourier-domain AS-OCT, with an axial resolution of 5 μm, is a promising method in the visualization of the structural details of the limbus. 4 – 6 The cross-sectional images of the limbus obtained by AS-OCT ( Fig. 1F ) reflect those seen in the histological section ( Fig.…”

Global Consensus on Definition, Classification, Diagnosis, and Staging of Limbal Stem Cell Deficiency

“…Anterior segment optical coherence tomography (AS-OCT) has emerged as an alternative imaging technique for LSCD, allowing for both noninvasive imaging of the ocular surface, including the limbus, and a larger field of view at the expense of resolution. [111][112][113][114][115] Although AS-OCT does not offer the same degree of resolution at the cellular level as IVCM does, AS-OCT may prove useful in measuring epithelial thickness, pannus depth, 116 assessing POV, limbal crypts, and the clear transition between the hyporeflective corneal epithelium and the hyperreflective conjunctival epithelium in the limbal region (i.e. cross-sections parallel and perpendicular to the limbus and en face sections).…”

The diagnosis of limbal stem cell deficiency

“…Spectral domain OCT has been reported to visualize the palisades of Vogt (Fig. 2C) [120]. This technique may enhance targeted limbal biopsies for transplantation [121].…”