Fluorescence-guided surgery (FGS) is a type of surgery that is performed with the guidance of fluorescence imaging. It has emerged as a new approach to detect the primary tumor, tumor margins, and metastatic lymph nodes. A major cause of tumor recurrence is the presence of residual tumor cells after surgery. Therefore, it is necessary to have a negative tumor margin to ensure a successful cancer resection. Despite widespread use of preoperative imaging techniques such as computed tomography (CT) scan, magnetic resonance imaging (MRI) scan, and positron emission tomography (PET) scan, the surgical margin positive rate failed to reflect remarkable improvement. The plausible reason for this failure is the reliance of surgeons on the visual inspection and intraoperative histopathological analysis of the frozen tissue specimen. FGS is now emerging as an effective tool to guide the surgical oncologists by generating real time fluorescence images of the surgical region. It reduces the possibility of recurrence of cancer. This book strives to provide a fair idea about the fluorescence-guided surgery and to help develop a better understanding of the latest advances related to this medical procedure. It is a vital tool for all researching and studying fluorescence-guided surgery as it gives incredible insights into the emerging trends and concepts.