A rapid and reversible skull optical clearing method for monitoring cortical blood flow

In vivo cortex optical imaging is of great important for revealing both structural and functional architecture of brain with high temporal-spatial resolution. To reduce the limitation of turbid skull, researchers had to establish various skull windows or directly expose cortex through craniotomy. Here we developed a skull optical clearing method to make skull transparent. Laser speckle contrast imaging technique was used to monitor the cortical blood flow after topical treatment with the optical clearing agents. The results indicated that the image contrast increased gradually, and then maintained at a high level after 15 min for adult mice, which made the image quality and resolution of micro-vessels nearly approximate to those of exposed cortex. Both the cortical blood flow velocity almost kept constant after skull became transparent. Besides, the treatment of physiological saline on the skull could make skull return to the initial state again and the skull could become transparent again when SOCS retreated it. Thus, we could conclude that the skull optical clearing method was rapid, valid, reversible and safe, which provided us available approach for performing the cortical structural and functional imaging at high temporal-spatial resolution.