Molecular Neurobiology Program, Department of Physiology and Neuroscience, Skirball Institute, New York University School of Medicine, New York, New York, USA.
Nat Protoc. 2010 Feb;5(2):201-8. doi: 10.1038/nprot.2009.222. Epub 2010 Jan 14.
Imaging neurons, glia and vasculature in the living brain has become an important experimental tool for understanding how the brain works. Here we describe in detail a protocol for imaging cortical structures at high optical resolution through a thinned-skull cranial window in live mice using two-photon laser scanning microscopy (TPLSM). Surgery can be performed within 30-45 min and images can be acquired immediately thereafter. The procedure can be repeated multiple times allowing longitudinal imaging of the cortex over intervals ranging from days to years. Imaging through a thinned-skull cranial window avoids exposure of the meninges and the cortex, thus providing a minimally invasive approach for studying structural and functional changes of cells under normal and pathological conditions in the living brain.
在活体大脑中对神经元、神经胶质细胞和脉管成像已成为理解大脑工作方式的重要实验工具。本文详细描述了一种通过活体小鼠颅骨开窗，利用双光子激光扫描显微镜（TPLSM）以高光学分辨率对皮质结构进行成像的方案。手术可以在 30-45 分钟内完成，之后可以立即进行图像采集。该过程可以重复多次，从而可以在从数天到数年的时间间隔内对皮质进行纵向成像。通过颅骨开窗进行成像可以避免脑膜和皮质暴露，从而为在活体大脑中研究正常和病理条件下细胞的结构和功能变化提供了一种微创方法。
