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(申请人提供)：超分辨率显微镜使生物学家能够绕过经典的衍射极限看到以前看不见的东西，并有望 广泛转变生物医学研究。然而，目前的方法只能提供有限的分辨率和多路复用能力，并且往往需要昂贵的仪器或专门的实验条件。因此，它们还没有在标准的生物学实验室中被广泛采用。我们提出了一种基于核酸荧光探针可编程自主闪烁的简单而健壮的超分辨率方法。与依赖于特殊荧光团的外部控制随机闪烁的现有技术不同，我们的技术利用具有规定亮度和闪烁频率的核酸(DNA/RNA)探针的可编程自主闪烁。这种对探测器闪烁行为的前所未有的分子控制水平将使成像技术能够提供高多路复用能力和分辨率。与目前的超分辨率方法不同，我们的可编程眨眼技术只需要标准仪器，具有广泛的适用性和潜在的遗传编码能力。这项技术的发展将把超分辨率成像的性能、可用性和适用性带到一个新的水平，并有助于改变不同生物医学领域的研究实践。