Abstract
Purpose :
Conjunctival goblet cells (CGCs) are specialized epithelial cells that secrete mucins to form the mucous layer of the protective tear film. They also contribute to ocular surface homeostasis through their immune tolerance functions. Although CGCs are an important biomarker for diagnosing ocular surface diseases, their information has been difficult to access due to the lack of rapid and noninvasive examination methods.
Methods :
We developed high-speed extended depth-of-field (EDOF) wide-field microscopy with surface tracking for non-contact real-time CGC imaging in human subjects. It is fluorescence microscopy using moxifloxacin, an ophthalmic antibiotic for CGC labeling. EDOF imaging was achieved by axially sweeping the focal plane with an electrically tunable lens and applying deconvolution. A novel long-range surface detection method was developed for real-time surface tracking during large-area imaging. The new EDOF microscope was demonstrated in rabbit model and human subject studies.
Results :
The performance was demonstrated first by the mosaic imaging of the whole conjunctiva in the normal rabbit model, ex vivo. Approximately 10 cm square in area was captured as 40 × 25 image patches. The CGC density map of the normal rabbit model was generated, and the spatial variations of CGC density were visualized. The EDOF microscope was applied to longitudinal imaging of ocular surface damage rabbit models induced by PI. Strip regions (5 mm × 1 mm) in the bulbar conjunctiva were imaged for CGC density analysis. The longitudinal changes in CGC density were observed, and these changes were analyzed in comparison with the accompanied dry eye evaluation results. The EDOF microscope with surface tracking was applied to three healthy adult subjects and demonstrated its ability of rapid large-area imaging of 10 mm square area in approximately 10s for reliable CGC density measurement
Conclusions :
Novel EDOF microscopy with surface tracking was developed for humans and its performance was demonstrated. This new imaging system could be useful for noninvasive CGC examination in humans, which could be valuable for precision diagnosis and optimal treatment of ocular surface diseases.
This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.