Coherent Super-Resolution Optical Microscopy for Enhanced Image Resolution and Speed

相干超分辨率光学显微镜可提高图像分辨率和速度

• 批准号: 1309041
• 负责人: Rajesh Menon
• 金额: $ 36万
• 依托单位: University of Utah
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-10-01 至 2019-09-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1309041&HistoricalAwards=false
• 关键词: Coherent; Super; Resolution; Optical; Microscopy

The objective of this proposal is to increase the resolving power and the image acquisition speed, of super-resolution fluorescence microscopy by about an order of magnitude (to 5-10nm). Resolution in conventional photoactivatable-localization microscopy is limited by the localization precision, which, in turn is proportional to 1/sqrt(N), where N is the number of collected photons. In the proposed approach, we can achieve localization precision that is proportional to 1/N. This is achieved by: (1) reduction in the effective size of the point-spread function (PSF) as a result of a novel grating system, (2) increase in the number of detected photons due to interference with a high intensity reference wave and (3) utilization of phase information in addition to the intensity of the signals using a novel optical correlator for optical filtering.The intellectual merit arises from the potential acquisition of fluorescence images with macro-molecular (sub-5nm) resolution enabled by a relatively simple addition to conventional confocal microscopes. The transformative nature of the proposed technique is a result of the higher resolution and faster imaging, which will elucidate the fundamental interactions between proteins and sub-cellular nanostructures with unprecedented spatial and temporal resolution. Such mechanistic understanding will prove to be essential for future advancements in biology.The broader impacts of this project include (1) training of the scientific workforce via an innovative, inter-disciplinary course, (2) recruitment of under-represented students via a hands-on demonstration module to be utilized at 3 large outreach events, and (3) widespread dissemination via commercialization of the subject technology.

本提案的目的是将超分辨率荧光显微镜的分辨率和图像采集速度提高约一个数量级（至5-10nm）。传统的光激活定位显微镜的分辨率受到定位精度的限制，而定位精度又与1/sqrt(N)成正比，其中N是收集到的光子数量。在提出的方法中，我们可以实现与1/N成正比的定位精度。这是通过：(1)减少点扩散函数（PSF）的有效尺寸作为一个新的光栅系统的结果，(2)增加检测光子的数量由于高强度参考波的干扰和(3)利用相位信息除了信号的强度使用一个新的光学相关器进行光学滤波。智能优点在于，通过在传统共聚焦显微镜上相对简单的添加，可以获得具有大分子（低于5nm）分辨率的荧光图像。所提出的技术的变革性质是更高分辨率和更快成像的结果，这将以前所未有的空间和时间分辨率阐明蛋白质和亚细胞纳米结构之间的基本相互作用。这种机械的理解将被证明对生物学未来的发展至关重要。该项目的更广泛影响包括：(1)通过创新的跨学科课程培训科学工作者；(2)通过在3个大型外展活动中使用的实践示范模块招募代表性不足的学生；(3)通过主题技术的商业化进行广泛传播。