Wide-field in vivo confocal microscopy of meibomian gland acini and rete ridges in the eyelid margin

PURPOSE. In vivo confocal microscopy (IVCM) has been widely used to evaluate changes in the meibomian glands (MGs) in response to age and disease. This study examined the structures described as MGs using wide-field IVCM and laser scanning confocal microscopy (LSCM) in situ and characterized their spatial distribution and localization relevant to the eyelid margin. METHODS. IVCM was performed on 30 subjects aged 18 to 38 to visualize structures in the eyelid margin. Size, shape, and distribution characteristics were measured, and individual frames were montaged into wide-field images. Structures observed on IVCM were then visualized using LSCM of whole-mount and cryosectioned cadaver eyelids stained with Nile red, mucin-1 (MUC1), laminin-5, and 4 ^′ ,6-diamidine-2 ^′ -phenylindole dihydrochloride. RESULTS. The size, distribution, and staining patterns of the reflective structures seen on IVCM did not correspond to the MGs in cadaver eyelids. Instead, staining profiles indicated that these structures corresponded to the rete ridges present at the dermal–epidermal junction. Wide-field imaging showed a densely populated field of rete ridges with distinct size and shape characteristics depending on their location relative to the meibomian orifices. A distal shift of the mucocutaneous junction (MCJ) was evident in some eyelids. CONCLUSIONS. IVCM is unable to visualize MGs in the human eyelid margin due to light attenuation at that tissue depth. LSCM confirms that these structures are rete ridges located at the dermal–epidermal junction. Alterations in the structure of the dermal–epidermal junction within the eyelid margin indicate a shifting of the MCJ and may impact tear film dynamics.