Neuro-Electronics Research Flanders, Leuven, Belgium.
VIB, Leuven, Belgium.
Nat Protoc. 2021 Jul;16(7):3547-3571. doi: 10.1038/s41596-021-00548-8. Epub 2021 Jun 4.
Most brain functions engage a network of distributed regions. Full investigation of these functions thus requires assessment of whole brains; however, whole-brain functional imaging of behaving animals remains challenging. This protocol describes how to follow brain-wide activity in awake head-fixed mice using functional ultrasound imaging, a method that tracks cerebral blood volume dynamics. We describe how to set up a functional ultrasound imaging system with a provided acquisition software (miniScan), establish a chronic cranial window (timing surgery: ~3-4 h) and image brain-wide activity associated with a stimulus at high resolution (100 × 110 × 300 µm and 10 Hz per brain slice, which takes ~45 min per imaging session). We include codes that enable data to be registered to a reference atlas, production of 3D activity maps, extraction of the activity traces of ~250 brain regions and, finally, combination of data from multiple sessions (timing analysis averages ~2 h). This protocol enables neuroscientists to observe global brain processes in mice.
大多数脑功能涉及到分布区域的网络。因此，对这些功能的全面研究需要评估整个大脑；然而，对行为动物的全脑功能成像仍然具有挑战性。本方案描述了如何使用功能超声成像来跟踪脑血流动力学，对清醒固定头部的小鼠进行全脑活动的跟踪。我们描述了如何使用提供的采集软件（miniScan）设置功能超声成像系统，建立慢性颅窗（手术时间：~3-4 小时）并以高分辨率（每脑切片 100×110×300µm 和 10Hz，每次成像大约需要 45 分钟）对与刺激相关的全脑活动进行成像。我们还提供了可将数据注册到参考图谱、生成 3D 活动图、提取约 250 个脑区活动轨迹的代码，最后可将多个时间段的数据进行组合（时间分析平均约 2 小时）。该方案使神经科学家能够在小鼠中观察到全局脑过程。
