CUBIC Protocol Visualizes Protein Expression at Single Cell Resolution in Whole Mount Skin Preparations

The CUBIC protocol enabled visualization of protein expression at single-cell resolution in whole mount skin preparations of mice, preserving tissue integrity and allowing detailed 3D imaging. It used aminoalcohol-based chemical cocktails to render skin tissue transparent while maintaining protein integrity, facilitating the study of basal and proliferating keratinocytes, sebaceous glands, and other structures. This method proved valuable for assessing skin anatomy, pathology, and regenerative processes, particularly in genetically modified mouse lines, and highlighted its utility in studying abnormal epidermal phenotypes and skin diseases. Despite limitations like antibody compatibility and IT resource needs, the protocol offered significant advancements over traditional 2D sectioning and other 3D techniques.