Super-resolution imaging via programmable autonomous blinking

通过可编程自主闪烁进行超分辨率成像

• 批准号: 8549370
• 负责人: Peng Yin
• 金额: $ 69.02万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-25 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8549370
• 关键词: Adopted; Area; Behavior; Binding; Biological; Biomedical Research; Blinking; Complex; DNA; DNA Probes; Devices; Docking; Engineering; Equipment; Fluorescence; Fluorescent Probes; Frequencies; Image; Imaging Techniques; Individual; Label; Life; Methods; Microscopy; Molecular; Nanostructures; Nucleic Acid Probes; Nucleic Acids; Optics; Performance; RNA; RNA Probes; Research; Resolution; Scheme; Techniques; Tubulin; Work; base; cellular engineering; density; fluorescence imaging; fluorophore; imaging probe; innovation; instrumentation; novel strategies; programs; public health relevance; reconstruction; research in practice; success; synthetic construct; technique development; usability

DESCRIPTION (provided by applicant): Super-resolution microscopy enables biologists to see the previously invisible by circumventing the classical diffraction limit, and holds promise to broadly transform biomedical research. However, current methods offer only limited resolution and multiplexing power, and tend to require either expensive instrumentation or specialized experimental conditions. They thus have not yet been widely adopted in standard biological labs. We propose a simple and robust super-resolution method based on the programmable autonomous blinking of a nucleic acid fluorescent probe. Unlike existing techniques that rely on the externally controlled stochastic blinking of special fluorophores, our technique utilizes programmable autonomous blinking of a nucleic acid (DNA/RNA) probe with prescribed brightness and blinking frequency. This unprecedented level of molecular control on the blinking behavior of the probe will enable an imaging technique that offers high multiplexing power and resolution. Unlike current super-resolution methods, our programmable blinking technique only requires standard instrumentation and is widely applicable and potentially genetically encodable. Development of this technique will bring the performance, usability, and applicability of super-resolution imaging to a new level, and help transform research practice in diverse biomedical fields.

描述（由申请人提供）：超分辨率显微镜使生物学家能够通过规避经典衍射限制来查看以前的看不见，并持有承诺 广泛地改变生物医学研究。但是，当前方法仅提供有限的分辨率和多重功率，并且倾向于需要昂贵的仪器或专门的实验条件。因此，它们尚未在标准生物实验室中广泛采用。我们提出了一种基于核酸荧光探针的可编程自主眨眼的简单稳健的超分辨率方法。与现有的技术依赖于特殊荧光团的外部控制随机眨眼的技术不同，我们的技术利用了具有亮度和闪烁频率的核酸（DNA/RNA）探针的可编程自主眨眼。对探针闪烁行为的这种前所未有的分子控制水平将实现一种成像技术，该技术具有高多重能力和分辨率。与当前的超分辨率方法不同，我们可编程的闪烁技术仅需要标准仪器，并且广泛适用并且潜在地具有遗传编码。该技术的开发将使超分辨率成像的性能，可用性和适用性提高到一个新的水平，并有助于改变不同生物医学领域的研究实践。