Abstract Advances in light microscopy, digital image processing, and the development of a variety of powerful fluorescent probes present expanding opportunities for investigating the nervous system, from synaptic spines to networks in the brain. Addressing the need for intensive research training in this field, Cold Spring Harbor Laboratory will offer the annual three-week course on Imaging Structure and Function in the Nervous System, as part of its postgraduate summer training program in neurobiology (2020-2024). This intensive laboratory and lecture course will provide participants with the theoretical and practical knowledge to utilize emerging imaging technologies. The primary focus of the course will be on in vivo applications of light microscopy, particularly functional imaging with genetically encoded calcium indicators. Methods taught will include multi-photon and light-sheet microscopy, and combination of imaging with optogenetics. Lectures by leading experts will progress through basic concepts to presentation of cutting-edge methods. Students will learn the fundamentals of optics and microscopy, as well as the use of different types of cameras, laser scanning systems, in vivo preparations and image processing and analysis software. A strong emphasis is placed on building exercises that allow students to develop an understanding of basic principles, while also introducing them to a variety of state-of-the-art commercial systems. Robust experimental design and data collection/management strategies are integral features emphasized throughout the course. The lead instructors are chosen on the basis of their contributions to and knowledge of the field. In turn, they invite lecturers who have made significant contributions in their fields to give up-to-the-minute reports on the underlying technology and their current research. The trainees are chosen by the course faculty from larger pools of applicants and range from graduate students to senior investigators and research physicians. This intensive research training program enrolls a diverse cadre of researchers who can immediately transfer techniques and concepts learned during the course to their own research projects and interests, while embedding the trainees in a collaborative network of researchers interested in using advanced light microscopy techniques to the study of brain function and dysfunction. The faculty are encouraged to present relevant methodological approaches in the broader context of brain function and mental health research. Methods and concepts taught in the course are disseminated to the wider research community through the publication of laboratory manuals and online resources.