Optimising the method for visualising mouse meibomian gland using eyelid whole-mount lipid staining

Abstract

Black opaque pigment residing in the eyelids of C57BL/6 inbred mice and other related mice hampers detailed whole-mount macroscopic observation of the meibomian gland. Owing to this inherent problem, studying mouse meibomian gland histology is and has been mainly performed by standard tissue-sectioning method, which allows detection of cross-sectioned image of the meibomian gland without significant interference from the interstitial pigments. However, appearance of tissue section of the meibomian gland is variable between sections. The chief advantage of whole-mount histological analysis, in comparison, is that it enables examination of the whole structure of the meibomian glands, not sections thereof, allowing accurate (non-section-biased) quantitative investigation of their morphology. In the present study, we optimized the method for visualizing mouse meibomian gland using eyelid whole-mount lipid staining. We prepared bleached eyelid samples and made them transparent. By doing so, we visualized the entire meibomian gland structure across the eyelid (see Fig. 1).