Development of spectroscopic Single-Molecule Localization Microscopy

项目摘要 最近的一个重大科学突破是超分辨率光学纳米镜的出现。 纳米尺度光学成像目前在生物医学科学中起着至关重要的作用， 在大分子尺度上的细胞结构和功能的信息。然而，这一潜力 技术可以大大提高。我们建议开发和优化一类新的光谱 超分辨率成像工具，称为光谱单分子定位显微镜或sSMLM。 它将使生物医学研究人员首次实现具有分子特异性的多色成像， 亚10纳米的空间分辨率，基本上具有无限数量的可区分分子。我们将开发 sSMLM采用模块化光学设计和开源数据处理软件。独特的光谱 分析光学模块将与现有的商业SMLM系统兼容， 改性新的硬件和软件开发以及优化的实验方案将 确保广泛的生物医学研究社区最大限度地访问sSMLM。我们预计sSMLM 将推动基础生物学研究中超分辨率成像的范围超越目前的静态 空间分析只有少数（<4）分子。

PROJECT SUMMARY One of the major recent scientific breakthroughs was the emergence of super-resolution optical nanoscopy. Nanoscale optical imaging currently plays a crucial role in biomedical sciences and has provided invaluable information about the cellular structure and function at the macromolecular scale. However, the potential of this technology can be immensely enhanced. We propose to develop and optimize a new class of spectroscopic super-resolution imaging tool, referred to as spectroscopic single-molecular localization microscope or sSMLM. It will allow biomedical researchers, for the first time, to achieve multi-color imaging with molecular specificity at sub 10-nm spatial resolution with principally unlimited number of distinguishable molecules. We will develope the sSMLM with modular optical designs and open-source data processing software. The unique spectral analysis optical module will be compatible with existing commercial SMLM systems with minimal or no system modification. The new hardware and software development together with optimized experimental protocols will ensure maximal accessibility of sSMLM by the broad biomedical research community. We anticipate that sSMLM will push the envelope of super-resolution imaging in fundamental biological research beyond current static spatial analysis of only few (<4) molecules.