细胞分辨水平的猕猴脑光学成像方法研究

The brain is the organ we understand the least.The rhesus macaque, as a non-human primate model animal, has a special value in the study of advanced brain cognitive function and human brain disease.The neural circuit is the structural and functional foundation of the brain to perform the functions. To understand the working mechanism of the brain, we have to study the whole neural circuit. Due to the brain volume of macaque is 200 times larger than that of the mouse brain, there is still no method to obtain the cell resolved connectional map of brain of macaque in circuit level. This project intends to use virus-mediated fluorescent protein-labeled macaque brain as a sample. I propose to study high-quality embedding methods for large-volume macaque brain samples to solve the problem of fluorescence quenching and autofluorescence enhancement in sample embedding. I propose to investigate cutting method for large sample to solve the problem that the precision and cutting speed of brain samples can not be balanced at the same time. I propose to study optical tomography with high space bandwidth products to solve the contradiction between the imaging resolution, signal-to-noise ratio and imaging speed of data acquisition. Finally, we will establish the imaging method of neural circuit in macaque brain at a single cell resolution. We will provide a new tool for future acquisition of fine connection altas of macaque whole-brain. Our method also can be used for brain disease research and advanced brain function mechanism research using primate model.

脑是人类认识最少的器官，猕猴作为非人灵长类模式动物在高级脑认知功能和人脑疾病研究方面具有独特的价值。神经环路是脑行使各项职能的结构与功能基础，完整环路信息的获取是理解大脑本质工作机理的基本前提。由于猕猴脑体积是鼠脑的200倍以上，目前仍缺乏工具对猕猴脑神经环路进行细胞分辨水平连接信息的获取。本项目拟以病毒介导的荧光蛋白标记的猕猴脑作为样本，通过研究针对猕猴脑大体积样本的高质量包埋方法，解决样本包埋中荧光蛋白淬灭和自发荧光提高的问题；通过研究猕猴脑大体积样本的切削方法，解决大体积的猕猴脑样本切削时精度和速度不能兼顾的问题；通过研究高空间带宽积的光学层析成像方法，解决猕猴脑大体积样本成像分辨率、信噪比与成像速度不可兼得的难题。最终建立针对猕猴脑神经环路在细胞分辨水平的成像方法，为将来获取猕猴全脑精细连接图谱提供新的工具，也可用于以灵长类动物为模型的脑疾病研究和高级脑功能机制的研究中。

脑是人类认识最少的器官，猕猴作为非人灵长类模式动物在高级脑认知功能和人脑疾病研究方面具有独特的价值。神经环路是脑行使各项职能的结构与功能基础，完整环路信息的获取是理解大脑本质工作机理的基本前提。由于猕猴脑体积是鼠脑的200倍以上，目前仍缺乏工具对猕猴脑神经环路进行细胞分辨水平连接信息的获取。本项目以病毒介导的荧光蛋白标记的猕猴脑作为样本，通过研究针对猕猴脑大体积样本的高质量包埋方法，解决样本包埋中荧光蛋白淬灭和自发荧光提高的问题；通过研究猕猴脑大体积样本的切削方法，解决大体积的猕猴脑样本切削时精度和速度不能兼顾的问题；通过研究高空间带宽积的光学层析成像方法，解决猕猴脑大体积样本成像分辨率、信噪比与成像速度不可兼得的难题。建立了针对猕猴脑神经环路在细胞分辨水平的成像方法，为将来获取猕猴全脑精细连接图谱提供新的工具。