小鼠双光子激光扫描显微镜活体成像窗口构建技术

Construction technologies of in vivo imaging window of mice for two-photon laser scanning microscopy

双光子显微成像技术以其组织器官穿透性深、光漂白及光毒性低等独特优势,成为小动物活体光学高分辨显微成像的重要工具;但是活体状态下,由于动物的呼吸、心跳以及一些组织非自主的蠕动等使观察视野无法固定,导致图像失焦,降低实验的可重复性。成像视窗以其支持原位、实时观察及长期成像等优势,已成为双光子活体成像的重要辅助手段之一。本文主要介绍了小鼠不同脏器活体成像固定装置的设计、成像视窗的构建方法以及成像窗口在实际应用中需注意的问题等,旨在为使用者提供更多的方法参考,解决活体成像图像稳定度低、清晰度差等问题,拓宽实验种类,提高观察深度和精度。

Two-photon microscopy imaging technology is an important tool for in vivo optical high-resolution microscopy imaging of small animals because of its unique advantages such as deep penetration into tissues and organs,low photobleaching and low phototoxicity.However,in the living state,due to the animal's breathing,heartbeat,and involuntary peristalsis of some tissues,the observation field cannot be fixed,which results in image defocus and reduces the repeatability of the experiment.The imaging window has become one of the important auxiliary methods for two-photon in vivo imaging with its advantages of supporting in-situ,real-time observation,and long-term imaging.Herein the design of in vivo imaging fixtures for different organs in mice,the construction methods of imaging windows,and the problems that need to be paid attention to in the actual application of imaging windows are mainly introduced,aiming to provide more method references for users,solve the problems of low stability and poor definition of in vivo imaging images,broaden the types of experiments,and improve the depth and accuracy of observation.