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

Intraoperative optical imaging is essential for surgical precision and patient safety, but current systems present anatomical and fluorescence information separately, causing delays and increasing cognitive load. A unified system for simultaneous visualization is critically needed to enhance guidance and efficiency. To develop and validate a multispectral imaging system that overcomes fragmented intraoperative imaging by simultaneously capturing and integrating white light reflectance and multiple fluorescence signals in real-time, enhancing surgical precision and safety. The system integrates two synchronized color cameras with optimized optical components, including custom filters and a beam splitter, tailored for clinically relevant fluorophores protoporphyrin IX (PpIX), fluorescein sodium, and indocyanine green (ICG). Advanced image processing techniques enhance visualization accuracy. Performance was evaluated using phantoms simulating clinical conditions. The system provided real time, simultaneous visualization of white light and multiple fluorescence signals without mode switching. Fluorescence emissions from PpIX, fluorescein sodium, and ICG were accurately separated using linear unmixing algorithms. Reflectance images showed high color fidelity, with Delta E values indicating imperceptible to barely perceptible differences compared to a conventional camera. Latency was minimal, ensuring immediate visual feedback suitable for intraoperative use. This multispectral imaging system addresses the limitations of fragmented intraoperative imaging by enabling continuous, real time, integrated visualization of anatomical and fluorescence information. It streamlines workflow, reduces cognitive load, and supports more precise interventions. By leveraging components similar to conventional surgical microscopes, it offers a practical, cost effective solution aligned with clinical needs.