Super-resolution microscopy through single moleculelocalization at cryogenic temperature

通过低温下单分子定位的超分辨率显微镜

• 批准号: 390890664
• 负责人: Professor Dr. Jörg Enderlein
• 金额: --
• 依托单位: III. Physikalisches Institut - Biophysik
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2017
• 资助国家: 德国
• 起止时间: 2016-12-31 至 2021-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/390890664?language=en
• 关键词: Super; resolution; microscopy; through; single

Super-resolution microscopy has revolutionized optical microscopy. In particular, methods based on stochastic fluorophore blinking, such as SOFI, or on single-molecule localization, such as PALM or STORM, have become widely popular. We propose to combine SOFI and PALM with cryo-fluorescence microscopy to enable imaging of shock-frozen samples with high conservation of structural detail and to open the door to correlative cryogenic optical and electron microscopy. A major bottleneck towards these goals is the lack of suitable fluorescent markers able to efficiently photo-switch at cryogenic temperatures, in particular photo-transformable fluorescent proteins (PTFPs). The core goal of our project is to design and characterize PTFPs optimized for cryo-SOFI and cryo-PALM, and to modify our existing cryo-fluorescence microscope for localizing single molecules with extreme precision and allowing cryo-CLEM studies. This research will contribute a new boost in the super-resolution field, of major interest to integrated structural and cell biology.

超分辨率显微镜彻底改变了光学显微镜。特别是，基于随机荧光团闪烁的方法，如SOFI，或基于单分子定位的方法，如PALM或STORM，已经变得广泛流行。我们建议将联合收割机SOFI和PALM与低温荧光显微镜相结合，以使冲击冷冻样品的成像具有高度的结构细节保护，并为相关的低温光学和电子显微镜打开大门。实现这些目标的主要瓶颈是缺乏能够在低温下有效地光开关的合适的荧光标记物，特别是可光转化的荧光蛋白（PTFP）。我们项目的核心目标是设计和表征针对cryo-SOFI和cryo-PALM优化的PTFP，并修改我们现有的冷冻荧光显微镜，以极精确地定位单分子并进行cryo-CLEM研究。这项研究将为超分辨率领域带来新的推动力，这对整合结构和细胞生物学具有重大意义。

项目成果

Fluorescence polarization filtering for accurate single molecule localization

• DOI: 10.1063/5.0009904
• 发表时间: 2020-06-01
• 期刊: APL PHOTONICS
• 影响因子: 5.6
• 作者: Nevskyi, Oleksii;Tsukanov, Roman;Enderlein, Joerg
• 通讯作者: Enderlein, Joerg