ABSTRACT This Academic-Industrial Partnership (AIP) will further the clinical development of fluorescence guided surgery (FGS) through the utilization of the FDA exploratory Investigational New Drug (eIND) pathway. While recent advances in both imaging agents and technology for FGS has been impressive, significant limitations remain in detecting low dose administration of imaging agents and obtaining true molecular contrast between the tumor and normal surrounding tissues. The basis of this application and the underlying tenet of the proposed AIP is that a cost-effective, risk-diluted approach to clinical translation of imaging methodologies is needed in order to realize the promise of FGS. Utilization of our previously developed bench-to-clinic pipeline stands to accelerate surgical oncology to revolutionize both the procedures that are possible and the surgical outcomes that will result. In the first funding cycle our AIP implemented a low-cost testing pipeline for ABY-029, a fluorescent epidermal growth factor receptor-targeted synthetic peptide. We established the necessary protocols to bring ABY-029 from pre-clinical animal models into three Phase 0 clinical trials using the FDA eIND pathway (eIND #122681). We will build upon our previous success to apply for Investigational New Drug (IND) status of ABY-029 for economical, investigator-led Phase II testing of detection accuracy in glioma and head and neck cancers. Sim- ultaneously, we will utilize the pipeline to achieve eIND approval of paired-agent imaging. The proposed AIP between Dartmouth (Engineering and Medical Schools), Affibody AB, LI-COR, and DoseOptics brings together 4 partners who have the intellectual property (IP), expertise and infrastructure to develop, test and advance molecular-targeted fluorescent tracers and imaging systems for surgical guidance. Clinical translation and testing of FGS will be achieved through the completion of the following aims: Aim 1 - Advance near-microdose ABY-029 FGS for resection of primary high-grade glioma in a Phase 2 trial; Aim 2 - Adapt FGS system for minimally invasive resection of head and neck cancers for Phase 2 testing; and Aim 3 - Leverage the Dartmouth fluorescent agent development pipeline to bring quantitative molecular paired- agent imaging (PAI) to Phase 0 trial using a cocktail of ABY-029 and IRDye 680LT carboxylate (pABY-IR680). The translational INNOVATION will include advancement of ABY-029 into Phase II clinical trials, first-in-human implementation of paired-agent imaging for true molecular contrast via the established bench-to-clinic eIND pipe- line, and the integration of a highly sensitive, multi-channel near infrared surgical camera imaging system capa- ble of imaging in full surgical lights. The SIGNIFICANCE of this AIP will be to demonstrate improved margin detection for surgical resection of glioma and head and neck cancers using a low-cost, efficient testing pipeline.