Abstract
Tissue clearing enables deep imaging of long biological structures using light microscopy approaches. Protocols such as iDISCO are not currently available for human skin. Here, we present Skin-iDISCO+, a tissue-clearing and labeling protocol for morphometric analysis of human cutaneous vasculature. We describe steps for processing and pre-treating tissue, immunostaining, clearing, and imaging using confocal and light-sheet fluorescence microscopy. We then detail procedures for the reconstruction of networks and the extraction of morphometric parameters of the skin vasculature. For complete details on the use and execution of this protocol, please refer to Renier et al.1.