Significance: Cerebral blood flow (CBF) regulation at neurovascular coupling (NVC) plays an important role in normal brain functioning to support oxygen delivery to activating neurons. Therefore, studying the mechanisms of CBF adjustment is crucial for the improved understanding of brain activity.

Aim: We investigated the temporal profile of hemodynamic signal change in mouse cortex caused by neural activation and its variation over cortical depth.

Approach: Following the cranial window surgery, intrinsic optical signal imaging (IOSI) was used to spatially locate the activated region in mouse cortex during whisker stimulation. Optical microangiography (OMAG), the functional extension of optical coherence tomography, was applied to image the activated and control regions identified by IOSI. Temporal profiles of hemodynamic response signals obtained by IOSI and OMAG were compared, and OMAG signal was analyzed over cortical layers.

Results: Our results showed that the hemodynamic response to neural activity revealed by blood flow change signal signal through IOSI is slower than that observed by OMAG signal. OMAG also indicated the laminar variation of the response over cortical depth, showing the largest response in cortical layer IV.

Conclusions: Overall, we demonstrated the development and application of dual-modality imaging system composed of IOSI and OMAG, which may have potential to enable the future investigations of depth-resolved CBF and to provide the insights of hemodynamic events associated with the NVC.