Oblique Light Sheet Fluorescence Microscopy With Surface Tracking for High-Speed Large-Area Imaging of Conjunctival Goblet Cells.

Abstract

Conjunctival goblet cells (CGCs) are specialized mucin-secreting epithelial cells, playing key roles for ocular surface homeostasis. Their examination is important for diagnosing various ocular surface disorders. However, existing imaging modalities have limitations in examining CGCs over large conjunctival regions. In this study, we present an oblique light sheet fluorescence microscopy (OLSFM) system integrated with real-time surface tracking for high-speed, large-area imaging of CGCs. The system incorporates an obliquely oriented imaging arm, providing high tolerance to variations in the sample surface. Continuous lateral scanning with pulsed light sheet illumination enables rapid large-area imaging. Additional real-time surface tracking maintains focus during the lateral scanning over the curved conjunctival surface. OLSFM successfully imaged an 8 mm² conjunctival area within 0.5 seconds in animal models, highlighting its potential for rapid CGC examination and the precise diagnosis of ocular surface diseases.