Surgical preparation of rats and mice for intravital microscopic imaging of abdominal organs

•Surgical approach depends on organ of interest and how it's presented on the stage.•Maintenance of homeostatic physiologic state is critical for reproducible data.•Proper aseptic surgical technique is paramount with survival procedures.•Peri- and post-operative animal management designed to optimize serial imaging outcomes.•Long term longitudinal studies may require the implementation of imaging windows.

Intravital microscopy is a powerful research tool that can provide insight into cellular and subcellular events that take place in organs in the body. However, meaningful results can only be obtained from animals whose physiology is preserved during the process of microscopy. Here I discuss the importance of preserving the overall state of health of the animal, methods of anesthesia, surgical techniques for intravital microscopy of various abdominal organs, methods to maintain and monitor the physiology of the animal during microscopy and associated peri- and post-operative recovery considerations.