This study is aimed to understand sensory nerve endings’ functional organization and morphology in the adult mouse cornea. Free nerve endings are essential for the sensory transduction of noxious and innocuous stimuli. However, their functional organization still needs to be fully understood. To investigate this, we used GCaMP6s-based in vivo imaging of calcium signals evoked by menthol and capsaicin in corneal nerve endings of 5-6 week-old C57 mice (Fig. 1A). We then acquired 3D confocal images of their morphological structure and examined the contribution of voltage-gated calcium channels (VGCC) to the calcium signal evoked by menthol.The results show that corneal nerves activated only by menthol had a higher proportion of ramified (55.6%) and complex (44.5%) nerve endings compared to those responding only to capsaicin (28.6% ramified, 0.0% complex) or to both drugs (25.0% ramified, 6.3% complex) (Fig. 1B). Blocking VGCC with 50µM benidipine led to a reversible reduction of the ∆F/F0 peak in menthol-responding nerve endings (29.5 ± 7.5%, p<0.001) (Fig. 1C). This reduction was observed in both intraepithelial nerve endings (36.2 ± 4.5%, p<0.001) and subbasal fibers (28.3 ± 2.4%, p<0.001) (Fig. 1D).These findings suggest that corneal cold-sensing nerve endings have a more complex morphology than polymodal nociceptor nerve endings. Furthermore, results suggest that voltage-gated calcium channels may play a role in the excitability of cold-sensing nerve endings.