Rapid Discrimination of High-Grade Prostate Cancer Using Label-Free Fluorescence Lifetime Measurements.

Purpose: Histologic evaluation of prostatic needle biopsies is essential for prostate cancer (PCa) diagnosis and treatment planning, yet tissue targeting remains suboptimal despite MRI-guided Bx procedures. This pilot study investigates the use of label-free Fluorescence Lifetime Imaging (FLIm) for real-time biopsy guidance. Using ex vivo specimens, we assess FLIm's preliminary efficacy in discriminating malignant from benign prostate tissue. Materials and Methods: Twenty patients undergoing prostate biopsy were enrolled. FLIm measurements were performed immediately after sample collection using a custom fiber-optic probe. Optical parameters from 4 spectral bands associated with distinct endogenous fluorophores including structural proteins and metabolic cofactors (e.g. NADH, FAD) were extracted and labeled based on histological annotation. Data were analyzed to characterize tissue-type differences and train and evaluate a classifier to distinguish malignancy. Results: Separation between benign tissue and Gleason grade ≥4 PCa was achieved using just 2 of 56 FLIm-derived parameters. A Support Vector Machine classifier using all parameters achieved a ROC of 0.88 in identifying grade 4 PCa. A reduced lifetime value in the NADH-associate band, likely due to increased free NADH from upregulated glycolysis, supports the biochemical basis for optical differentiation. Conclusions: FLIm shows significant potential for high-grade PCa identification. The single-fiber approach requires minimal modification for integration into current biopsy tools, supporting its feasibility for clinical translation.