In vivo imaging of central nervous system fluid spaces using synchrotron radiation-based micro computed tomography

Current approaches to in vivo imaging of the mouse central nervous system (CNS) do not offer a combination of micrometer resolution and a whole-brain field of view. To address this limitation, we introduce an approach based on synchrotron radiation-based hard X-ray micro computed tomography (SR$\mu$CT). We performed intravital SR$\mu$CT acquisitions of mouse CNS fluid spaces at three synchrotron radiation facilities. Imaging was conducted on both anesthetized free-breathing and ventilated animals, with and without retrospective cardiac gating. We achieved whole-brain imaging at 6.3 $\mu$m uniform voxel size, observed the distribution of cerebrospinal fluid (CSF) contrast agent over time and quantified choroid plexus movement. SR$\mu$CT bridges the gap between multiphoton microscopy and magnetic resonance imaging, offering dynamic imaging with micrometer-scale resolution and whole-organ field of view. Intravital SR$\mu$CT will play a crucial role in validating and integrating hypotheses on CSF dynamics and solute transport by providing unique data that cannot be acquired otherwise.