Yang Guang, Pan Feng, Chang Paul C, Gooden Frank, Gan Wen-Biao
Department of Anesthesiology, New York University School of Medicine, New York, NY, USA.
Methods Mol Biol. 2013;1010:35-43. doi: 10.1007/978-1-62703-411-1_3.
Transcranial two-photon microscopy allows long-term imaging of neurons, glia, and vasculature in the intact cortex of living animals. So far, this technique has been primarily used to acquire images in anesthetized animals. Here, we describe a detailed protocol for high-resolution two-photon imaging of neuronal structures in the cortex of awake head-restrained mice. Surgery is done within 1 h in anesthetized mice. After animals recover from anesthesia, two-photon imaging can be performed multiple times over minutes to days, allowing longitudinal studies of synaptic plasticity and pathology without the complication induced by anesthesia reagents.
经颅双光子显微镜可对活体动物完整皮层中的神经元、神经胶质细胞和脉管系统进行长期成像。到目前为止,该技术主要用于在麻醉动物身上获取图像。在此,我们描述了一种用于清醒头部固定小鼠皮层神经元结构高分辨率双光子成像的详细方案。手术在麻醉小鼠体内1小时内完成。动物从麻醉中恢复后,可在数分钟至数天内多次进行双光子成像,从而能够对突触可塑性和病理学进行纵向研究,而不会受到麻醉剂引发的并发症影响。