title: CUBIC Protocol Visualizes Protein Expression at Single Cell Resolution in Whole Mount Skin Preparations.
abstract: The skin is essential for our survival. The outer epidermal layer consists of the interfollicular epidermis, which is a stratified squamous epithelium covering most of our body, and epidermal appendages such as the hair follicles and sweat glands. The epidermis undergoes regeneration throughout life and in response to injury. This is enabled by K14-expressing basal epidermal stem/progenitor cell populations that are tightly regulated by multiple regulatory mechanisms active within the epidermis and between epidermis and dermis. This article describes a simple method to clarify full thickness mouse skin biopsies, and visualize K14 protein expression patterns, Ki67 labeled proliferating cells, Nile Red labeled sebocytes, and DAPI nuclear labeling at single cell resolution in 3D. This method enables accurate assessment and quantification of skin anatomy and pathology, and of abnormal epidermal phenotypes in genetically modified mouse lines. The CUBIC protocol is the best method available to date to investigate molecular and cellular interactions in full thickness skin biopsies at single cell resolution.
authors: The School of Medical Sciences, University of New South Wales Australia; Neuroscience Research Australia.
authors: Huazheng
authors: H
authors: Liang
authors: The School of Medical Sciences, University of New South Wales Australia.
authors: Bassem
authors: B
authors: Akladios
authors: The School of Medical Sciences, University of New South Wales Australia.
authors: Cesar P
authors: CP
authors: Canales
authors: Biomedical Imaging Facility, University of New South Wales Australia.
authors: Richard
authors: R
authors: Francis
authors: The School of Medical Sciences, University of New South Wales Australia.
authors: Edna H
authors: EH
authors: Hardeman
authors: The School of Medical Sciences, University of New South Wales Australia; a.beverdam@unsw.edu.au.
authors: Annemiek
authors: A
authors: Beverdam
mesh_terms: D000818:Animals; D001706:Biopsy / Q000379:methods; D018859:Hair Follicle / Q000166:cytology / Q000000981:diagnostic imaging / Q000378:metabolism; D015603:Keratinocytes / Q000166:cytology / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018613:Microscopy, Confocal / Q000379:methods*; D012038:Regeneration / Q000502:physiology; D012627:Sebaceous Glands / Q000166:cytology / Q000000981:diagnostic imaging / Q000378:metabolism; D059010:Single-Cell Analysis / Q000379:methods*; D012867:Skin / Q000166:cytology* / Q000000981:diagnostic imaging / Q000378:metabolism*; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism; D046509:Tissue Culture Techniques / Q000379:methods