报告题目：使用vDISCO对完整器官和成年啮齿动物体进行全景成像
时 间：2019年4月22日10:30
地 点：光电国家研究中心 A101
报 告 人：Cai Ruiyao（蔡瑞瑶）, 德国慕尼黑大学卒中和痴呆研究所
邀 请 人：骆海明 教授
【个人简介】
Cai Ruiyao（蔡瑞瑶）出生于中国浙江，5岁随父母搬到意大利米兰。 她在意大利米兰-比可卡大学于2011、2014年分别获得生物技术学士和硕士学位，其硕士论文研究软脑膜脑血管是如何影响脑中风。自2015年1月至今，她在德国慕尼黑大学卒中和痴呆研究所的Ali Ertürk博士实验室攻读博士学位，其研究方向是开发新的组织清除技术，以探索由创伤性脑损伤、脊髓损伤等脑损伤引发的变化。目前，Cai Ruiyao共发表SCI论文9篇，其中以第一作者在Nature Neuroscience、Nature Methods杂志上发表SCI论文各一篇。
Ruiyao Cai was born in Zhejiang, China. At the age of 5 she moved to Milan, Italy with her parents and here she grew up. She obtained her BSc and MSc in Biotechnology at the University of Milan-Bicocca, Italy and during her Master‘s thesis she worked in the laboratory of Neurobiology on the role of leptomeningeal cerebral vessels in influencing the stroke outcome. After a period as intern at the University of Oxford and as research fellow in the lab of Neurobiology of the University of Milan-Bicocca, where she continued in the stroke field, she moved to Munich for her PhD studies in Neuroscience under the GSN graduate school. Currently, she is completing her doctoral program in Dr. Ali Ertürk's lab at the Institute for Stroke and Dementia Research, LMU, in Munich. Her research projects focus on the development of new tissue clearing technologies to explore changes triggered by brain injuries such as traumatic brain injury and spinal cord injury. Ruiyao has published 9 refereed international papers in top journals including first-authored papers in Nature Neuroscience (2018) and Nature Methods (2016).
【摘要】 大多数疾病的影响并不局限于特定的身体区域，因此需要用系统生物学方法来研究全身水平的疾病。组织清除方法彻底改变了标准组织学，允许对器官和生物进行成像而无需切片。到目前为止，DISCO清除方法已实现最高水平的组织透明度，且它们还能与抗体标记相兼容。然而，由于有机溶剂最终可以淬灭内源荧光信号，机体内生物荧光信息的可靠检测和定量仍然是一个挑战。为了克服这个问题，我们开发了“vDISCO”，它利用基于纳米抗体的全身免疫标记系统来增强荧光蛋白的信号，可以对致密和坚硬组织进行成像和永久保存。vDISCO允许我们在完整透明小鼠中成像和量化亚细胞细节。使用vDISCO，我们在亚细胞分辨率下可视化成年小鼠的第一个全身神经元图谱。接下来观察小鼠中枢神经系统创伤后的变化，我们发现躯干周围神经末梢退化。此外，使用vDISCO我们观察到骨髓和脑膜之间的短血管连接，其在中风时充满白细胞。总之，我们的方法是分析疾病对整个生物体影响的有力工具。
The effects of most diseases are not confined to a particular body region, therefore a systems biology approach is needed to study diseases at a whole-body level. Tissue clearing methods revolutionized standard histology, allowing the imaging of organs and organisms without sectioning. So far, the highest level of tissue transparency has been achieved by DISCO clearings. It has also been shown that they are compatible with antibody labeling. Nevertheless, owing to the fact that organic solvents can eventually quench the endogenous fluorescent signal, the reliable detection and quantification of the biological fluorescent information in all body districts have still represented a challenge. To overcome this issue, we developed vDISCO", which exploits a nanobody-based whole-body immunolabeling system to enhance the signal of fluorescent proteins, to preserve it permanently and to image through dense and hard tissues such as bones and skin. vDISCO allowed us to image and quantify subcellular details in intact transparent mice. Using vDISCO we visualized the first whole-body neuronal map in adult mice at subcellular resolution. Next, we screened whole mice for changes after CNS trauma and we found degeneration of peripheral nerve terminals in the torso. Moreover, using vDISCO we observed short vascular connections between skull marrow and meninges, which were filled with leucocytes upon stroke. Taken together, our method represents a powerful tool to analyze effects of diseases in whole organisms.
