Simultaneous real-time multispectral fluorescence and reflectance imaging for enhanced intraoperative guidance

Intraoperative fluorescence imaging enhances surgical precision, but current systems require switching between anatomical and fluorescence views, disrupting workflow and forcing surgeons to mentally overlay separate images. We developed a dual-sensor multispectral imaging system that uniquely enables simultaneous visualization of full-color anatomy and multiple fluorescence signals (protoporphyrin IX, fluorescein sodium, and indocyanine green) in real-time, a capability not available in existing surgical platforms. By employing custom optical filters and linear unmixing algorithms, the system captures all imaging modes concurrently, providing perfectly co-registered anatomical and fluorescence information in a single unified view. Phantom validation demonstrated accurate fluorophore separation while maintaining color fidelity for tissue visualization. This approach eliminates mode-switching delays and mental image fusion, streamlining surgical workflow with a practical platform designed for clinical integration.