Evaluating the Performance of Hyperspectral Imaging Endoscopes: Mitigating Parameters Affecting Spectral Accuracy.

Hyperspectral imaging (HSI) is increasingly used in studies for medical applications as it provides both structural and functional information of biological tissue, enhancing diagnostic accuracy and clinical decision-making. Recently, HSI cameras (HSICs) have been integrated with medical endoscopes (HSIEs), capturing hypercube data beyond conventional white light imaging endoscopes. However, there are currently no cleared or approved HSIEs by the U.S. Food and Drug Administration (FDA). HSI accuracy depends on technologies and experimental parameters, which must be assessed for reliability. Importantly, the reflectance spectrum of a target can vary across different cameras and under different environmental or operational conditions. Thus, before reliable clinical translation can be achieved, a fundamental question must be addressed: can the same target yield consistent spectral measurements across different HSI systems and under varying acquisition conditions? This study investigates the impact of eight parameters-ambient light, exposure time, camera warm-up time, spatial and temporal averaging, camera focus, working distance, illumination angle, and target angle-on spectral measurements using two HSI techniques: interferometer-based spectral scanning and snapshot. Controlled experiments were conducted to evaluate how each parameter affects spectral accuracy and whether normalization can mitigate these effects. Our findings reveal that several parameters significantly influence spectral measurements, with some having a more pronounced impact. While normalization reduced variations for most parameters, it was less effective at mitigating errors caused by ambient light and camera warm-up time. Additionally, normalization did not eliminate spectral noise resulting from low exposure time, small region of interest, or a spectrally non-uniform light source. From these results, we propose practical considerations for optimizing HSI system performance. Implementing these measures can minimize variations in reflectance spectra of identical targets captured by different cameras and under diverse conditions, thereby supporting the reliable translation of HSI techniques to clinical applications.