Hochauflösendes konfokales Fluoreszenz-Mikroskop mit STED-Technologie

采用 STED 技术的高分辨率共焦荧光显微镜

• 批准号: 414012445
• 金额: --
• 依托单位: Technische Universität Braunschweig
• 依托单位国家: 德国
• 项目类别: Major Research Instrumentation
• 财政年份: 2018
• 资助国家: 德国
• 起止时间: 2017-12-31 至 无数据
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/414012445?language=en
• 关键词: Hochaufl; sendes; konfokales; Fluoreszenz; Mikroskop

Functional and structural changes at synapses (plasticity) have been shown to underlie learning and memory processes. A very precise balance between plasticity and stability of synapses and thereby of the mature CNS network is crucial to ensure learning and memory formation. Thus, it is of crucial interest to analyse changes in the architecture of dendritic spines, the site of most excitatory synapses and to correlate them to functional changes. In addition, synaptic localization of molecules involved in synaptic transmission and plasticity is of extreme interest. In particular, it is of prime importance to analyse activity-dependent changes in the localization of plasticity-promoting (e.g. BDNF) or plasticity-limiting (e.g. Nogo-A) proteins and their consequences on spine/synapse structure. However, the imaging methods used so far suffer from intrinsic limitations that do not allow to resolve changes occurring below optical resolution. Combining confocal-based super-resolution (HyVolution) and Nanoscopy (Stimulated Emission Depletion, STED) will allow us to analyse structural plasticity upon learning, molecular mechanisms regulating it and its functional consequences. The combination of HyVolution and STED will allow us to analyse changes in size and shape of synaptic structures as well as protein/mRNA localization relative to the synapses under physiological (learning) or pathological conditions (Alzheimer, mental retardation, neuroinflammation).

突触的功能和结构变化(可塑性)已被证明是学习和记忆过程的基础。在突触的可塑性和稳定性以及成熟的中枢神经系统网络之间保持非常精确的平衡，对于确保学习和记忆的形成至关重要。因此，分析树突棘结构的变化是至关重要的，树突棘是大多数兴奋性突触的位置，并将它们与功能变化联系起来。此外，参与突触传递和可塑性的分子的突触定位也引起了极大的兴趣。特别是，分析可塑性促进蛋白(如BDNF)或可塑性限制蛋白(如Nogo-A)定位的活性依赖变化及其对脊柱/突触结构的影响是至关重要的。然而，到目前为止使用的成像方法受到固有的限制，不允许解决发生在光学分辨率以下的变化。结合基于共焦的超分辨率(HyVolution)和纳米显微镜(受激发射耗尽，STED)将使我们能够分析学习时的结构可塑性，调节它的分子机制及其功能后果。HyVolution和STED的结合将使我们能够分析突触结构的大小和形状的变化，以及在生理(学习)或病理条件(阿尔茨海默病、智力低下、神经炎症)下相对于突触的蛋白质/mRNA定位。