Cellular and Molecular Imaging of Synaptic Vesicle Dynamics

我们正在进行一项成像计划，利用最新的超材料超透镜技术来构建超透镜纳米镜，以获得优越的时间和空间分辨率，并研究脑突触传递和细胞吞噬/胞吐过程中的膜动力学。这种超透镜由特殊设计和微细加工的结构元件组成，能够以亚衍射限制的空间分辨率放大近场图像，并实时将高分辨率图像投射到远场(姚等人，2008，Science，321：930；Liu，et et.阿尔，2007，科学，315：1686；方等人。Al，2005，Science，308：534-7)。与EM、STED(Hell，2007，Science，316：1153-8)和Storm/Palm(Huang，等，2008，Science，319：810-813；Bates等)等其他成像技术相比，这种形式的光学纳米显微镜具有实时光学成像(视频率)和纳米空间分辨率的一些独特优势。等人，2007年，《科学》，1749-53；赫斯等人，2007年，《美国国家科学院院刊》第104卷，17370-5页；贝齐格等人。Al，2006，Science 313：1642)。在孙成教授(现为西北大学校外合作者)和张X.教授(加州大学伯克利分校)的带领下，超透镜相关技术正在迅速发展。 在从TRANS-NIH成像计划获得设备拨款后，我们一直在NIH开发我们的成像设备，并致力于解决建模和纳米制造问题，以改进超透镜的设计和建造。在这个项目年，我们还利用我们在原子力显微镜(AFM)和各种光学显微镜平台上的多模式仪器的相关经验，将牛嗜铬细胞(固定的或活的，或转染了各种荧光蛋白结构)作为囊泡运输模型系统进行了研究。我们仍在为基础生物学和医学的亚细胞膜动力学的突破性研究而努力。

We are undertaking an imaging initiative to build a hyperlens nanoscope, using newly advanced meta-material hyperlens technology, to achieve superior temporal and spatial resolution and to investigate membrane dynamics during brain synaptic transmission and cellular endocytosis/exocytosis in general. The hyperlens consists of specially designed and micro-fabricated structure elements that can magnify near-field images at sub-diffraction-limited spatial resolution and project the high resolution image at far-field in real time (Yao, et al, 2008, Science, 321:930; Liu, et. al, 2007, Science, 315:1686; Fang, et. al, 2005, Science, 308:534-7). This form of optical nanoscope possesses some unique advantages of both real-time optical imaging (at video rate) and nano-metric spatial resolution, in comparison with other imaging techniques such as EM, STED (Hell, 2007, Science, 316: 1153-8), and STORM/PALM (Huang, et al., 2008, Science, 319:810-813; Bates, et. al, 2007, Science 317, 1749-53; Hess, et al, 2007, PNAS 104, 17370-5; Betzig, et. al, 2006, Science 313:1642). Hyperlens-related technology is under rapid development led by the groups of Prof. Cheng Sun (extramural collaborator now at Northwestern University) and Prof. X. Zhang (UC Berkeley). After winning an equipment grant from the trans-NIH imaging initiative, we have been developing our imaging setup at NIH and are working to resolve modeling and nano-manufacturing issues toward improved hyperlens design and construction. In this project year, we have also investigated bovine chromaffin cells (fixed or live, or transfected with various fluorescent protein constructs) as a vesicular trafficking model system, by leveraging our related experience on multimodal instrumentation around atomic force microscopy (AFM) and various optical microscopy platforms. We are still working toward breakthrough investigations of sub-cellular membrane dynamics for both fundamental biology and medicine.