Fluorescence-guided surgery (FGS) is defined as a medical imaging technique that uses a fluorescent dye or a near-infrared emitting light source to identify anatomic structures during surgical procedures. In 2020 alone, over 1200 academic articles were published on the topic of fluorescence-guidance surgery, a sign that this modality is making significant inroads into surgical practice. The use of near-infrared imaging and FGS is a rapidly growing modality, allowing surgeons to see more intraoperatively, enhance surgical precision, and improve surgical decision-making and patient outcomes. This manual provides a comprehensive, state-of-the art review of this field and will serve as a valuable resource for clinicians, surgeons and researchers with an interest in fluorescence-guided surgery, guiding patient management and stimulating investigative efforts. After initial chapters discussing the history of FGS and the current platforms and devices, it presents the most up-to-date data regarding the use of FGS in multiple surgical fields - colorectal, hepatic, endocrine, reconstructive, pediatric, among others - as well as in the treatment of specific conditions such as burns. Chapters are generously illustrated with full-color figures and intraoperative photographs, and selected chapters include video segments. Access to a comprehensive resource such as this is currently limited by the relatively new inroads that fluorescence-guided technology has made into surgery. The SAGES Manual of Fluorescence-Guided Surgery fills this gap in the literature.