Metabolics Underlying Dynamic Optical Contrast Imaging for Head & Neck Cancer Margin Determination

Purpose/Objective(s) To describe the metabolics underlying cancer and cancer margin detection of lifetime fluorescence in Dynamic Optical Contrast Imaging (DOCI). Materials/Methods Cell lines Tu686, OKF, FADU, and RH2 were cultured in and sequentially treated with mitochondrial poisons oligomycin, FCCP, then rotenone + antimycin A. Treated cells, untreated cells, and vehicle control were plated with powdered magnesium at 50,000 cell density and imaged with the DOCI machine. These same cell lines then were plated in a 96 well culture microplate with the same mitochondrial poisons as above, and analysis was run via a Seahorse XF96 assay, yielding in vitro metabolic data. Redox ratio was calculated for the cell lines after mitochondrial poisons were treated, and were compared to data from the Seahorse assay. Results Redox ratio as measured from the DOCI machine correlate with changes in the oxygen consumption rate of the cell, as measured via Seahorse, after administration of oligomycin, FCCP, and rotenone with antimycin A. In both DOCI and Seahorse assay, average values decreased after administration of oligomycin, increased after adding FCCP, and again decreased when adding rotenone + antimycin A. Conclusion DOCI has been utilized for cancer margin detection, and this in vitro study demonstrates the underlying mechanism of detecting specific metabolic shifts, by comparing DOCI redox ratios to Seahorse assay metabolic data.