Pancreatic cancer is the third leading cause of cancer-related deaths in the United States. Up to 20% of patients are candidates for resection, which is currently the only potentially curative treatment option. However, pancreatic cancer patients have high rates of recurrence after resection due in part to cells left behind at the surgical margin. Fluorescence guided surgery (FGS) has emerged as a method to improve surgical resection. For pancreatic cancer patients eligible for surgical resection, intraoperative imaging would serve two purposes. For patients with resectable disease, image-guidance could help delineate tumor tissue to achieve complete resection. Of critical value, fluorescence guided surgery (FGS) could be used to identify locally metastatic disease, which could spare the patient from major, unnecessary surgery and move directly to other treatments. Recent studies demonstrated that MUC16 is overexpressed in 60-80% of pancreatic cancers, but this biomarker has not yet been explored for FGS of pancreatic cancer. Herein, we describe the validation and development of a near infrared fluorescence antibody that recognizes MUC16 for surgical imaging.