研究人员报道了一种2.17g的微型三光子显微镜（m3PM），该显微镜具有在高散射状态下成像时最大限度地收集荧光的配置。研究人员人员通过成像整个皮层和海马背侧CA1的钙活性来证明其能力，高达1.2毫米深度，激光功率安全。它还能够在自由移动的小鼠在单球到达任务中检测后顶皮层第6层神经元的感觉运动行为相关活动。此外，m3PM成像能够研究自由行为动物的深层皮层和皮层下结构的神经机制，如海马背侧和纹状体背侧。
据介绍，在深层组织多光子显微镜中，荧光光子的扩散和散射，而不是来自焦点的弹道发射，一直是一个混杂因素。
附：英文原文
Title: Miniature three-photon microscopy maximized for scattered fluorescence collection
Author: Zhao, Chunzhu, Chen, Shiyuan, Zhang, Lifeng, Zhang, Dong, Wu, Runlong, Hu, Yanhui, Zeng, Fengqingyang, Li, Yijun, Wu, Dakun, Yu, Fei, Zhang, Yunfeng, Zhang, Jue, Chen, Liangyi, Wang, Aimin, Cheng, Heping
Issue&Volume: 2023-02-23
Abstract: In deep-tissue multiphoton microscopy, diffusion and scattering of fluorescent photons, rather than ballistic emanation from the focal point, have been a confounding factor. Here we report on a 2.17-g miniature three-photon microscope (m3PM) with a configuration that maximizes fluorescence collection when imaging in highly scattering regimes. We demonstrate its capability by imaging calcium activity throughout the entire cortex and dorsal hippocampal CA1, up to 1.2mm depth, at a safe laser power. It also enables the detection of sensorimotor behavior-correlated activities of layer 6 neurons in the posterior parietal cortex in freely moving mice during single-pellet reaching tasks. Thus, m3PM-empowered imaging allows the study of neural mechanisms in deep cortex and subcortical structures, like the dorsal hippocampus and dorsal striatum, in freely behaving animals.
DOI: 10.1038/s41592-023-01777-3
Source: https://www.nature.com/articles/s41592-023-01777-3
Nature Methods：《自然—方法学》，创刊于2004年。隶属于施普林格·自然出版集团，最新IF：47.99
官方网址：https://www.nature.com/nmeth/
投稿链接：https://mts-nmeth.nature.com/cgi-bin/main.plex
本期文章：《自然—方法学》：Online/在线发表
