snRNA-seq of murine tarsal plates

The eyelid is a complex environment containing, containing epidermal, hair follicles, and conjunctival cells closely adjacent to the Meibomian glands (MGs), making it challenging to isolate pure MG cells or to identify MG contributions in bulk RNA-seq analyses. To analyze transcription in MG subpopulations, and to overcome difficulties in isolating cytoplasmic mRNAs from lipid-rich acinar cells, we utilized snRNA-seq which allows for identification of cell sub-populations within a complex mix. Overall design: The dorsal portion of the eyelids and the surrounding tissues near the tarsal plates were removed using a scalpel. The tarsal plates were digested with collagenase A and the MGs were dissected using forceps. Nuclei were analyzed from the pooled samples of four 8-week-old and four 21-month-old male mice using two replicates for each age (16 mice analyzed in total).