High-speed optofluidic imaging is an emerging interdisciplinary technology that seamlessly integrates high-speed optical imaging and microfluidics, which is capable of realizing high-resolution, high-throughput, high-content imaging, and quantitative analysis of biological organisms in high-speed and complex fluid environments. High-speed optofluidic imaging has exhibited promising application potential in bioenergy, food science, drug screening, disease diagnosis, and many other fields. In this review, we introduce the basic principles, key techniques, and recent advances of high-speed optofluidic imaging. The future prospects and challenges of high-speed optofluidic imaging are also discussed.