Hou Steven S, Yang Joyce, Kwon Yeseo, Pian Qi, Tang Yijing, Dauphinais Christine A, Calvo-Rodriguez Maria, Khatib Mirna El, Vinogradov Sergei A, Sakadzic Sava, Bacskai Brian J
Department of Neurology, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02129.
Athinoula A. Martinos Center for Biomedical Imaging, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02129.
bioRxiv. 2025 Jan 23:2025.01.22.634409. doi: 10.1101/2025.01.22.634409.
Multiphoton microscopy serves as an essential tool for high-resolution imaging of the living mouse brain. To facilitate optical access to the brain during imaging, the cranial window surgery is commonly used. However, this procedure restricts physical access above the imaging area and hinders the direct delivery of imaging agents and drugs. To overcome this limitation, we have developed a cannula delivery system that enables the implantation of a low-profile cannula nearly parallel to the brain surface at angles as shallow as 8 degrees, while maintaining compatibility with multiphoton microscopy. To validate this approach, we perform direct infusion and imaging of various fluorescent cell markers in the brain. Additionally, we successfully demonstrate tracking of degenerating neurons over time in Alzheimer's disease mice using Fluoro-Jade C. Furthermore, we show longitudinal imaging of brain tissue partial pressure of oxygen using a phosphorescent oxygen sensor. Our developed technique should enable a wide range of new longitudinal imaging studies in the mouse brain.
多光子显微镜是对活体小鼠大脑进行高分辨率成像的重要工具。为便于成像过程中对大脑进行光学观察，通常采用颅骨开窗手术。然而，该手术限制了成像区域上方的物理通路，阻碍了成像剂和药物的直接递送。为克服这一局限性，我们开发了一种套管输送系统，该系统能够以低至8度的角度植入几乎与脑表面平行的薄型套管，同时保持与多光子显微镜的兼容性。为验证该方法，我们对大脑中的各种荧光细胞标记物进行了直接输注和成像。此外，我们成功地使用氟玉髓C对阿尔茨海默病小鼠中退化神经元随时间的变化进行了追踪。此外，我们使用磷光氧传感器对脑组织氧分压进行了纵向成像。我们开发的技术应能在小鼠大脑中开展广泛的新型纵向成像研究。
