Dermal fibroblasts play a central role in the production and remodeling of dermal extracellular matrix. Their spatially distinct subpopulations exhibit different functions during skin homeostasis, wound healing, and other diseases such as fibrosis and cancer. Here, we present a protocol for detecting 3D dermal fibroblast cellular dynamics in mice without fixing the tissue using an intravital imaging technique. We describe steps for preparing, labeling, and mounting mice. We then describe detailed procedures for intravital imaging, recovery, and quantification. For complete details on the use and execution of this protocol, please refer to Rognoni et al..