A Long-Term Cranial Window for High-Resolution Photoacoustic Imaging

In this study, we introduce the design, fabrication, and assessment of an optically and acoustically transparent, long-term and biocompatible cranial window for high-resolution photoacoustic microscopy of rat cerebral cortex.The cranial window is fabricated with a polydimethylsiloxane (PDMS) layer bonded with a glass ring (outer diameter: 8 mm, inner diameter: 5 mm) via air plasma cleaning. A detailed comparison of image quality was performed with the implantation of cranial windows using different thicknesses of the PDMS film, and the cover glass. In addition, long-term in vivo monitoring of rat cerebral cortex was conducted to evaluate the stability of the cranial window. Furthermore, we successfully applied this window for longitudinal photoacoustic imaging in freely moving rats.Based on a detailed evaluation, the cranial window fabricated with PDMS has a better imaging quality compared with a conventional cover-glass-based cranial window. The optimal film thickness is 50 μm considering the elastic deforming capability of PDMS. The cranial window maintained good quality for 21 and 12 days in anesthetized and free moving rats, respectively.The cranial window has a good imaging quality for both anesthetized and behaving rats, enabling long-term, high-resolution, and steady photoacoustic imaging of cerebral vasculatures.Based on the studies of both anesthetized and behaving rats, the proposed cranial window has the potential to be used in the longitudinal in vivo study of chronic brain diseases in freely moving rodents.