本项课题旨在突破多尺度全脑连接图谱绘制所面临的神经环路显微成像技术瓶颈，发展适合于啮齿类和非人灵长类的新型神经元标记技术和脑组织透明技术，建立完备和实用的样品制备流程，实现微米分辨率高通量荧光显微成像技术的新突破，提出高效、快速的图像处理新方法。研究内容：（1）优化适合于猕猴全脑连接图谱绘制和新型脑组织透明技术的示踪剂组合策略，并发展高效的单纯疱疹病毒等新型示踪工具；（2）阐明脑组织透明技术荧光长时间保持的机理，并发展适合于非人灵长类的新型脑组织透明技术；（3）发展偏振光敏感光学相干断层成像等新成像技术；（4）建立基于逆向示踪标记的快速、可靠的自动化细胞识别和计数方法，以及基于偏振光敏感光学相干断层成像的高效、快速的微米尺度脑白质纤维束重建新方法。为绘制细胞类型特异的、基于即早基因表达和嗜神经病毒示踪等策略的全脑连接图谱提供新技术和新方法学。

The purpose of this study is to break through the bottleneck of microscopic imaging of neural circuits faced by the construction of a multi-scale whole-brain connectivity atlas, to develop new neural tracing and tissue optical clearing technology for rodents and nonhuman primates, to establish a comprehensive and practical procedure for tissue sample preparation, to achieve a new breakthrough in high-throughput fluorescence microscopic imaging technology at the micrometer scale, and to develop innovative and efficient image processing methods. Research contents: (1) To optimize the tracer combination strategy that is suitable for the construction of a macaque whole-brain connectivity atlas and for the new tissue optical clearing techniques, and to develop efficient new tracer tools, such as herpes simplex virus; (2) to elucidate the mechanism of long-term preservation of fluorescence in tissue optical clearing and to develop new tissue optical clearing techniques suitable for nonhuman primates; (3) to develop new imaging techniques such as polarization-sensitive optical coherence tomography; (4) to establish fast and reliable automated cell recognition and counting methods for retrograde neuronal tracing, as well as efficient and fast reconstruction of white matter tracts at the micrometer scale based on polarization-sensitive optical coherence tomography. This study will provide new technology and methodology for the construction of a whole-brain connectivity atlas that is a cell-specific and based on strategies such as immediately early gene expression and neurotropic virus tracing.