Development of spectroscopic Single-Molecule Localization Microscopy

项目总结 最近的重大科学突破之一是出现了超分辨率光学纳米显微镜。 纳米级光学成像目前在生物医学科学中发挥着至关重要的作用，并提供了无价的 关于大分子尺度上的细胞结构和功能的信息。然而，这一潜力 技术可以得到极大的提升。我们建议开发和优化一类新的光谱 超分辨率成像工具，简称光谱单分子定位显微镜或sSMLM。 这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 空间分辨率低于10纳米，可区分分子的数量基本不受限制。我们将发展 采用模块化光学设计和开源数据处理软件的sSMLM。独一无二的光谱 分析光学模块将与现有的商业SMLM系统兼容，系统最少或没有系统 修改。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获得sSMLM。我们期待着SSMLM 将使超分辨率成像在基础生物学研究中的极限超越目前的静态 仅对少数(&lt；4)分子进行空间分析。

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.