Intravital Multiphoton Imaging of the Mouse Ocular Surface: From the Central Cornea to the Limbus

Intravital multiphoton imaging enables the real-time visualization of distinct cell populations and also cellular dynamics in the ocular surface of transgenic mice expressing fluorescent proteins. In contrast to the relatively flat topography of skin tissue, the mouse cornea presents a steep curvature, which poses a challenge for peripheral imaging. Here, we demonstrate a time-lapse, single-cell resolution multiphoton microscopic imaging methodology that overcomes this limitation. Using distinct transgenic mouse strains, our approach allows for targeted visualization of diverse cell types including corneal limbal epithelial stem cells (LESCs), their subcellular structures, and nerve fibers across the entire cornea.