Bioimaging has advanced the field of nanomedicine, drug delivery, and tissue engineering by directly visualizing the dynamic mechanism of diagnostic agents or therapeutic drugs in the body. In particular, wide-field, planar, near-infrared (NIR) fluorescence imaging has the potential to revolutionize human surgery by providing real-time image guidance to surgeons for target tissues to be resected and vital tissues to be preserved. In this review, we introduce the principles of NIR fluorescence imaging and analyze currently available NIR fluorescence imaging systems with special focus on optical source and packaging. We also introduce the evolution of the FLARE intraoperative imaging technology as an example for image-guided surgery.