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.

跨大脑区域的神经元活动测量对于理解认知和运动过程（如注意力、决策制定和行动选择）的神经相关性非常重要。然而，允许进行蜂窝分辨率测量的技术非常昂贵，并且需要高度的技术专长，这限制了它们的广泛使用。基因编码指标的广域成像是一种高通量、经济高效且灵活的方法，用于测量具有高时间分辨率和皮质范围视野的特定细胞群的活动。在这里，我们概述了我们的协议，用于组装广域宏观设置、执行手术以准备完整的头骨和长期在表现良好的转基因小鼠中成像神经活动。此外，我们强调了一个利用新颖的处理管道，基于云的方法来分析大规模成像数据集。该协议针对正在寻求建立宏观显微镜、优化长期慢性成像和/或分析皮质范围神经元记录的手术程序的实验室。整个协议，包括宏观组装和校准、手术准备、成像和数据分析的步骤，总共需要 8 小时。它旨在供在成像方法方面专业知识有限或对行为过程中的高通量成像感兴趣的实验室使用。手术准备、影像学和数据分析，共需要8 h。它旨在供在成像方法方面专业知识有限或对行为过程中的高通量成像感兴趣的实验室使用。手术准备、影像学和数据分析，共需要8 h。它旨在供在成像方法方面专业知识有限或对行为过程中的高通量成像感兴趣的实验室使用。