Cold Spring Harbor Laboratory, Neuroscience, Cold Spring Harbor, NY, USA.
Department of Neurobiology, University of California, Los Angeles, Los Angeles, CA, USA.
Nat Protoc. 2021 Jul;16(7):3241-3263. doi: 10.1038/s41596-021-00527-z. Epub 2021 Jun 2.
Measurements of neuronal activity across brain areas are important for understanding the neural correlates of cognitive and motor processes such as attention, decision-making and action selection. However, techniques that allow cellular resolution measurements are expensive and require a high degree of technical expertise, which limits their broad use. Wide-field imaging of genetically encoded indicators is a high-throughput, cost-effective and flexible approach to measure activity of specific cell populations with high temporal resolution and a cortex-wide field of view. Here we outline our protocol for assembling a wide-field macroscope setup, performing surgery to prepare the intact skull and imaging neural activity chronically in behaving, transgenic mice. Further, we highlight a processing pipeline that leverages novel, cloud-based methods to analyze large-scale imaging datasets. The protocol targets laboratories that are seeking to build macroscopes, optimize surgical procedures for long-term chronic imaging and/or analyze cortex-wide neuronal recordings. The entire protocol, including steps for assembly and calibration of the macroscope, surgical preparation, imaging and data analysis, requires a total of 8 h. It is designed to be accessible to laboratories with limited expertise in imaging methods or interest in high-throughput imaging during behavior.
跨脑区神经元活动的测量对于理解注意力、决策和动作选择等认知和运动过程的神经相关性非常重要。然而，允许细胞分辨率测量的技术既昂贵又需要高度的技术专长，这限制了它们的广泛使用。基因编码指示剂的宽场成像技术是一种高通量、具有成本效益且灵活的方法，可以高时间分辨率和全皮层视野测量特定细胞群体的活动。在这里，我们概述了组装宽场显微镜设置、进行手术准备完整颅骨以及在行为活跃的转基因小鼠中进行慢性神经活动成像的方案。此外，我们还强调了一种利用新型基于云的方法分析大规模成像数据集的处理管道。该方案针对的是那些正在寻求构建显微镜、优化长期慢性成像手术程序和/或分析全皮层神经元记录的实验室。整个方案，包括显微镜的组装和校准、手术准备、成像和数据分析步骤，总共需要 8 小时。它旨在为成像方法经验有限或对行为期间高通量成像感兴趣的实验室提供帮助。
