新型宽场超分辨及超高速光学显微成像基础研究与应用

Optical imaging plays a critical role in the exploration and discovery of the unknown world. In many important applications, it not only needs to display in the space with the super resolution, but also needs to measure in the time with the ultra-high speed. The existing super-resolution or ultra-high speed optical imaging is limited to the compromise of spatial and temporal resolutions, and therefore it is difficult to simultaneously obtain the super-resolution and ultra-high speed optical imaging. To break the compromise of spatial and temporal resolutions in the optical imaging, we plan to develop a new wide-field super-resolution and ultra-high speed optical microscopy technique. Here, we will combine the structured illumination microscopy (SIM) and compressed ultrafast photography (CUP). In this new technique, we utilize the wide-field imaging of both SIM and CUP and the single shot and multi-frame imaging of CUP, and we also develop the new method of three-dimensional computational imaging. Finally, we hope to obtain the optical microscopy with the spatial resolution less than 100 nm and the frame number more than 1250 fps. Moreover, we will apply this technique to the measurement of biological subcellular dynamics, and realize the super-resolution and ultra-high speed imaging of spatial distribution, structure, function and interaction in subcellular organisms. Through this research project, it can provide a well-established tool to explore the ultrafast phenomena in the micro world under the nanoscale, and also can provide an important research foundation for the application studies in the subject areas of physics, chemistry, biology and medicine.

光学成像在人类探索和发现未知世界奥秘扮演至关重要的角色。在很多关键科学应用研究中，不仅需要在空间上超分辨显示，同时还需要在时间上超高速测量。现有超分辨或超高速光学成像受限于空间和时间分辨率的相互制约，难以获取既超分辨又超高速光学成像。为了打破光学成像空间和时间分辨率的这种对弈，本项目拟发展一种新型宽场超分辨及超高速光学显微成像技术，该技术结合了结构光照明显微SIM和压缩超高速成像CUP，利用SIM和CUP都是宽场成像特性以及CUP单次拍摄多帧成像优势，辅之以三维计算成像新方法，最终实现空间分辨率小于100纳米和成像帧率大于1250帧/秒的光学成像，并把该项新技术应用于生物亚细胞动力学测量，实现亚细胞结构生物体空间分布、结构、功能及相互作用的超分辨及超高速成像。该项目研究为探索纳米尺度下微观世界超快现象提供强有力的技术支撑，同时为物理、化学、生物、医学等学科领域研究和应用提供重要的研究基础。

在光学成像相关的很多关键科学应用研究中，不仅需要在空间上超分辨显示，同时还需要在时间上超高速测量。现有超分辨或超高速光学成像受限于空间和时间分辨率的相互制约，难以获取既超分辨又超高速光学成像。为了打破光学成像空间和时间分辨率的这种相互制约，本项目将宽场结构光照明显微与压缩超高速成像相结合，提出了多种新型宽场超分辨及超高速光学显微成像技术，同时开发了一系列高速超分辨成像新算法，最终实现了空间分辨率100纳米和成像帧率大于1200帧/秒的光学成像，并成功捕获微流控芯片中流动的荧光小球。在压缩超高速成像方面，我们发明了四维和五维成像新方法，提出了一系列提升图像重构质量的高性能算法，并在超快光场诊断领域获得重要应用。此外，我们还开展了宽场超分辨拉曼显微成像研究，为无标记超分辨成像在生物应用中提供了一条新的途径；分别开发了新型荧光染料和smURFP荧光蛋白等新型探针，并研究了其发光机理。本项目研究成果在超高速光学成像、生物微结构动力学观测、流体动力学研究等方面具有重要的应用前景。项目执行期间，共计发表研究论文54篇，其中高水平论文13篇（含Phys. Rev. Lett. 1篇，JACS 1篇，Angew. Chem. Int. Ed. 2篇，Adv. Photonics 2篇，ACS Photonics 2篇，Photonics Res. 3篇，Opto-Electron. Adv. 1篇，Sci. China Phys. Mech. 1篇）,申请发明专利11项，已授权发明专利6项，其中申请美国发明专利1项并已获批。项目负责人因其在光场调控领域的突出成就获得2019年上海市自然科学一等奖（2/4），入选2021年度上海市优秀学术/技术带头人和上海市普陀区领军人才，并担任Photonics Asia等国内外学术会议的专题组会委成员。