Purpose of review Intravital imaging with multiphoton microscopy enables the detailed study of dynamic cellular processes within functioning organs in living animals, in ways that would not otherwise be possible. It therefore represents a powerful tool in translational kidney research. In this article, we will discuss several new technical developments that have significantly increased the capabilities of kidney imaging. Recent findings Important contemporary advances in biomedical imaging technology include longer wavelength excitation lasers, far-red emitting fluorescent reporters, highly sensitive detectors, fluorescence lifetime measurements, adaptive optics, microendoscopes, high-throughput automated analysis algorithms and tissue clearing techniques. Several recent studies have utilized intravital microscopy to gain valuable new insights into important physiological and pathophysiological processes in the kidney, such as renal handling of albumin and the cellular pathogenesis of acute kidney injury in sepsis. Summary Major technological advances are rapidly expanding the frontiers of intravital microscopy, which is likely to play an increasingly important role in preclinical kidney research in the coming years.