CUBIC protocol visualizes protein expression at single cell resolution in whole mount skin preparations

Liang, Huazheng, Akladios, Bassem, Canales, Cesar P., Francis, Richard, Hardeman, Edna H., and Beverdam, Annemiek (2016) CUBIC protocol visualizes protein expression at single cell resolution in whole mount skin preparations. Journal of Visualized Experiments, 1 (114). e54401.

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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.

Item ID: 60722
Item Type: Article (Research - C1)
ISSN: 1940-087X
Keywords: cellular biology, issue 114, skin, epidermis, CUBIC, 3D, imaging, confocal, mouse
Funders: National Health and Medical Research Council of Australia (NHMRC)
Projects and Grants: NHMRC APP1062720
Date Deposited: 23 Oct 2019 12:36
FoR Codes: 06 BIOLOGICAL SCIENCES > 0699 Other Biological Sciences > 069999 Biological Sciences not elsewhere classified @ 100%
SEO Codes: 92 HEALTH > 9299 Other Health > 929999 Health not elsewhere classified @ 100%
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