Fast and high-fidelity in situ 3D imaging protocol for stem cells and niche components for mouse organs and tissues

Quantitative 3D imaging of organ-wide cellular and subcellular components is central for revealing and understanding complex interactions between stem cells and their microenvironment. Here, we present a gentle but fast whole-mount immunofluorescence staining protocol for 3D confocal microscopy (iFAST3D) that preserves the 3D structure of the entire tissue and that of subcellular struc-tures with high fidelity. The iFAST3D protocol enables reproducible and high -res-olution 3D imaging of stem cells and various niche components for many mouse organs and tissues.For complete details on the use and execution of this protocol, please refer to Sac,ma et al. (2019).

Automated summary

This study proposes a gentle but fast whole-mount immunofluorescence staining protocol that preserves the 3D structure of complex interactions between stem cells and their microenvironment. The protocol enables high-resolution 3D imaging of various mouse organs and tissues.