Significance: The practicality of optical methods detecting tissue optical contrast (absorption, elastic and inelastic scattering, fluorescence) for surgical guidance is limited by interferences from blood pooling and the resulting partial or complete inability to interrogate cortex and blood vessels.

Aim: A multispectral diffuse reflectance technique was developed for intraoperative brain imaging of hemodynamic activity to automatically discriminate blood vessels, cortex, and bleeding at the brain surface.

Approach: A manual segmentation of blood pooling, cortex, and vessels allowed the identification of a frequency range in hemoglobin concentration variations associated with high optical signal in blood vessels and cortex but not in bleeding. Reflectance spectra were then used to automatically segment areas with and without hemodynamic activity as well as to discriminate blood from cortical areas.

Results: The frequency range associated with low-frequency hemodynamics and respiratory rate (0.03 to 0.3 Hz) exhibits the largest differences in signal amplitudes for bleeding, blood vessels, and cortex. A segmentation technique based on simulated reflectance spectra initially allowed discrimination of blood (bleeding and vessels) from cortical tissue. Then, a threshold applied to the low-frequency components from deoxyhemoglobin allowed the segmentation of bleeding from vessels. A study on the minimum acquisition time needed to discriminate all three components determined that ∼25  s was necessary to detect changes in the low-frequency range. Other frequency ranges such as heartbeat (1 to 1.7 Hz) can be used to reduce the acquisition time to few seconds but would necessitate optimizing instrumentation to ensure larger signal-to-noise ratios are achieved.

Conclusions: A method based on multispectral reflectance signals and low-frequency hemoglobin concentration changes can be used to distinguish bleeding, blood vessels, and cortex. This could be integrated into fiber optic probes to enhance signal specificity by providing users an indication of whether measurements are corrupted by blood pooling, an important confounding factor in biomedical optics applied to surgery.