A Multimodal Hyperspectral and Laser Speckle Imaging Platform for Simultaneous Monitoring of Spinal Cord Oxygenation, Perfusion and Metabolism
Hyperspectral imaging (HSI) enables spatial mapping of haemoglobin oxygenation and cytochrome-c-oxidase (CCO) redox changes in exposed neural tissue. In this study, we present a technical upgrade of an existing HSI platform by the integration of laser speckle contrast imaging (LSCI), enabling contemporaneous assessment of oxygenation, oxidative metabolism, relative perfusion and a derived relative oxygen metabolic index in exposed spinal cord in a rat preclinical model. The upgraded system (hNIR+) uses a shared optical path and a single camera to acquire HSI and LSCI images from the same field of view using a sequential acquisition workflow. Hyperspectral images are acquired at 11 discrete wavelengths (600, 630, 665, 784, 800, 818, 835, 851, 868, 881, and 894 nm) to estimate changes in oxygenated haemoglobin (HbO2), deoxygenated haemoglobin (HHb) and oxidized CCO (oxCCO). LSCI is implemented using 632.8-nm coherent illumination to generate relative blood-flow index (rBFI) maps. The LSCI subsystem was validated using a controlled blood-flow phantom, where the LSCI-derived inverse speckle contrast index showed a strong positive linear relationship with pump-controlled flow. The upgraded platform was then evaluated in proof-of-concept observations of the rat spinal cord under normoxia, hypoxia and recovery conditions. Representative multimodal maps and ROI-based time-course analysis showed clear dynamic responses in HbO2, HHb, oxCCO and rBFI during oxygen challenges that were consistent with the expected physiological responses of the spinal cord during hypoxia, hyperoxia and subsequent recoveries. Overall, these results demonstrate that integrating LSCI into HSI extends the platform to multimodal and simultaneous monitoring of haemodynamic-metabolic coupling.