Simultaneous axial-multifocal imaging of neural and vascular dynamics in the brain of awake mice using two-photon microscopy and head-mounted OCT

Understanding brain function requires large-scale, high-speed imaging of both neuronal and vascular dynamics. However, traditional two-photon microscopy and head-mounted Optical Coherence Tomography (OCT) typically suffer from limited depth of field, which restricts effective multi-plane imaging for functional neuroscience studies. In this study, we employ custom diffraction optical elements to generate single or multiple needle-shaped beams that extend the axial focal depth. This allows us to achieve an axial energy distribution with a Rayleigh length up to 10 times greater than conventional setups. Additionally, by modulating the gradient optical field, we effectively counteract light attenuation with depth, providing a comprehensive optical platform to explore the functional and structural aspects of the brain in awake mice.