Intravital Microscopy Techniques to Image Wound Healing in Mouse Skin

The ability to visualize biological phenomenon has driven scientific interest and advancement over the centuries. Although many methods and assays provide a detailed snapshot of a physiology, the ability to track such processes in real time has expanded the breadth of questions that can be interrogated in the laboratory. Intravital Microscopy (IVM) is a dynamic and powerful way to investigate both the homeostatic and host response to either therapeutic or pathological intervention using live animals. In this technique, animal models, (often mice) are anesthetized, and the organ of interest surgically exteriorized. The animal containing fluorescent labels (either endogenous, or conjugated to antibodies/proteins) will then be placed on a high-powered laser scanning microscope, where the labeled cells or structures can be observed in their natural environment. Complex behavioral data and interactions can be captured in a temporal manner, providing a plethora of information that will help researchers make conclusions on a more systemic level, rather than isolating only part the response. As the technology advances, a greater number of imaging modality options can be utilized, and more diverse research questions can be addressed. The goal of this chapter is to highlight IVM as a technique and help instruct new users on how to choose the proper modalities, and by using imaging of a skin wound in mice as a model, provide troubleshooting strategies, technical advice, and considerations.