One-Photon/Two-Photon/Lifetime Confocal Microscope

描述（由申请人提供）：了解纳米尺度上生物和人工系统的特性不仅需要能够以精确的原子/分子水平控制来操纵、制造、合成和组装它们，而且还需要能够用最先进的纳米级光谱和成像工具来表征和探测它们。荧光显微镜为探测纳米级系统提供了许多优势。它是非侵入性的，并提供三维成像;它具有高灵敏度（低至单分子水平），并允许观察分子和细胞器特异性信号。我们建议收购一个组合的单光子/双光子/寿命配置（;al激光扫描显微镜（1 P/2 P/寿命CLSM），以支持在加州大学洛杉矶分校的加州纳米系统研究所（CNSI）进行的人工和生物纳米系统组件的研究。组合的1 P/2 P/Lifetime CLSM将允许以可实现的最高空间和时间分辨率表征生物和人工系统。这种先进的光学工具将特别有用：（1）研究生物大分子在活细胞中的动态相互作用;（2）在单个光学平面上以高分辨率显示多个荧光标记（光学平面切片）和从连续切片重建3D图像;（3）嵌入散射介质和组织中的多个荧光标记物的可视化和3D切片，具有高信噪比和最小的光漂白。 1 P/2 P/Lifetime CLSM将用于解决悬而未决的问题，涵盖化学，生物学，生理学和材料科学的广泛主题。我们建议在以下研究项目中使用1 P/2 P/Lifetime CLSM的先进性能：（1）活细胞中半导体探针的靶向，（2）电压依赖性离子通道构象变化的检测和（3）正常和营养不良肌纤维横管系统中结构和功能异常的检测。 1 P/2 P/Lifetime CLSM的获得将大大增强CNSI进行的纳米级科学以及我们对细胞中心功能的理解。

DESCRIPTION (provided by applicant): Understanding the properties of biological and artificial systems on the nanometer scale requires not only the ability to manipulate, fabricate, synthesize and assemble them with precise atomic/molecular level control, but also the ability to characterize and probe them with state-of-the-art nanoscale spectroscopic and imaging tools. Fluorescence microscopy offers many advantages for probing nanoscale systems. It is non-invasive and provides imaging in three dimensions; it has high sensitivity (down to the single molecule level), and allows the observation of molecular- and organelle-specific signals. We propose the acquisition of a combined One-Photon / Two-Photon / Lifetime Confo(;al Laser Scanning Microscope (1P/2P/Lifetime CLSM) to support research in artificial and biological nanoscale system assemblies conducted at the California Nano-Systems Institute (CNSI) at UCLA. The combined 1P/2P/Lifetime CLSM will allow the characterization of biological and artificial systems with the highest spatial and temporal resolutions achievable. This advanced optical tool would be particularly useful for: (1) The study dynamic interactions of biological macromolecules in living cells; (2) The visualization of multiple fluorescent markers at high resolution in a single optical plane (optical plane sectioning) and reconstruction of 3D images from consecutive sections; (3) The visualization and 3D sectioning of multiple fluorescent markers embedded in scattering media and tissues with high signal to noise and minimal photo-bleaching. The 1P/2P/Lifetime CLSM will be use to address outstanding questions covering a broad spectrum of topics in Chemistry, Biology, Physiology and Material Sciences. We propose to use the advanced performances of the 1P/2P/Lifetime CLSM in the following research projects: (1) The targeting of semiconductor nanocrystal probes in live cells, (2) The detection of conformational changes in voltage dependent ion channels and (3) The detection of structural and functional abnormalities in the transverse tubular system of normal and dystrophic muscle fibers. The acquisition of the 1P/2P/Lifetime CLSM would considerably enhance the nanoscale science performed at the CNSI and our understanding of the central functions of the cell.