Visualizing Cellular Dynamics and Protein Localization in 3D Collagen

Immune cells migrate and communicate through cell-to-cell interactions and cy-tokines to coordinate the specificity and timing of the immune response. While studying these events in cell culture are standard procedure, spatiotemporal dy-namics of cell-to-cell interactions within three-dimensional (3D) environments are critical in generating appropriate effector functions. Here, we present a detailed protocol to study cells within an all-in-one 3D collagen matrix that is amenable to live-cell microscopy and immunohistochemistry. This approach facilitates ana-lyses of dynamic cellular events in 3D settings.For complete details on the use and execution of this protocol, please refer to Koh et al. (2020).