In recent years, retinal research has benefited from major advances in optical imaging approaches. Investigations of the structural and functional organization of the vertebrate retina using live preparations have been facilitated by improvements in cell labeling methods, and by microscopy techniques that permit high-resolution of cells in vitro and in vivo. In particular, the generation of transgenic animals with fluorescently labeled retinal cells has permitted real-time visualization of cell generation, migration, differentiation and growth in the developing retina. Neuronal activity can also be examined by optical imaging using activity reporters directed to specific retinal cell types. Optical techniques such as multiphoton microscopy and total internal reflection fluorescence microscopy (TIRFM) have helped unravel the physiological properties and function of retinal cells. Here, we focus on the latest cell labeling methods that have proven highly useful in many aspects of retinal research. We also highlight several examples of how newly developed imaging technology itself has facilitated investigations that have advanced our understanding of retinal circuits and their development.