Fluorescent Labeling of Rat-tail Collagen for 3D Fluorescence Imaging

physical characteristics of numerous tissues within the body, namely the skin. While confocal imaging techniques such as fluorescence reflection and two-photon microscopy are able to visualize collagen fibrils during 3D imaging without fluorescence, other imaging modalities require direct conjugation of fluorescent dyes to collagen. Here we detail how to generate 3D collagen gels labeled with a fluorescent dye. Furthermore, we go through the steps required to reproducibly generate bright collagen hydrogels that are suitable for live cell 3D imaging techniques.