Breast cancer is the most common malignant tumor among women worldwide, seriously threatening their physical and mental health. Near-infrared fluorescence (NIRF) imaging technology has the characteristics of fast imaging speed, high sensitivity, no radiation and simple operation. It is expected to achieve in vivo and real-time intraoperative guidance to optimize accuracy and results, and has completely changed breast cancer surgery. This chapter introduces the application and research of NIRF in surgical fields such as the evaluation of breast-conserving surgery margins for breast cancer, sentinel lymph node localization, axillary reverse mapping (ARM), and reconstruction surgery, and discusses its limitations and advantages. With the advancement of NIRF imaging technology, it is likely to become a powerful clinical tool for accurate breast cancer surgery.