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用于大景深三维纳米分辨多分子追踪的衍射光学元件的设计制备和实验研究*
Design, fabrication, and experimental demonstration of a diffractive optical element with long depth of field for nanoscale three-dimensional multi-molecule tracking*
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摘要: 发展能实时检测完整细胞内多个生物分子随时空变化的单分子探测和追踪技术，对于研究生命过程的分子机理具有重要意义。在变形光栅多阶成像和双螺旋点扩散函数成像方法的基础上，基于波前编码的原理，提出将二者优化结合，获得全新的衍射光学元件，该器件同时具有多重平面成像和双螺旋点扩散函数成像的功能，旨在解决活细胞内单分子探测和追踪技术中的大景深探测难题。设计和制备了该器件，并基于该器件搭建了显微成像系统，实验模拟证明该衍射光学元件同时可实现轴向12μm 的探测范围，与理论设计结果相符合，从而有效扩大了显微镜系统的景深，证明了设计的可行性。Abstract: The development of real-time single-molecule detection and tracking technology in time and space for multiple bio-molecules in intact cells has important significance for the study on molecular behaviors in life processes. However, limited depth of field is the main drawback of conventional microscopy that prevents observation and tracking of multiple molecules in intact cells. Based on the principle of wavefront coding, the design and fabrication of a novel diffractive optical element (DOE) which combines a distorted diffraction grating with a double-helix point spread function (PSF) phase plate so that it can simultaneously perform multi-plane imaging with double-helix PSF and realize an extended depth of field is presented. Experiments have been carried out on a self-built microscopic system based on the novel DOE, showing that a depth of field can be up to 12 μm. Experimental results are in good agreement with the theoretical predictions, thus proving the feasibility of this method.
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