Zhang Cilong, Xu Ning, Tan Qiaofeng
State Key Laboratory of Precision Measurement Technology and Instruments, Department of Precision Instrument, Tsinghua University, Beijing 100084, China.
Biomed Opt Express. 2022 Oct 31;13(11):6113-6123. doi: 10.1364/BOE.473899. eCollection 2022 Nov 1.
Structured illumination microscopy (SIM) enables live-cell super-resolution imaging with wide field of view (FOV) and high imaging speed, but the illumination system is usually bulky. With the advantages of small structure and high efficiency, lattice patterns assisted by diffractive optical elements (DOEs) have been used for structured illumination in SIM. But it is still challenging to raise the spatial frequency of diffractive lattice patterns when using traditional DOE design method, and thus the super-resolution imaging performance is restricted. In this paper, we propose a novel design method for DOE to generate lattice patterns with spatial frequency close to the cut-off frequency. It is the first time to obtain a lattice pattern with such high spatial frequency by diffractive optics. Finally, the proposed SIM achieves a lateral resolution of 131 nm at 519 nm fluorescent light while maintaining an original size as a standard inverted fluorescence microscope by only inserting a single well-designed DOE in the illumination optical path, which may promote this compact SIM applied in super-resolution imaging field.
结构光照明显微镜(SIM)能够实现具有宽视野(FOV)和高成像速度的活细胞超分辨率成像,但照明系统通常体积庞大。具有结构小和效率高的优点,由衍射光学元件(DOE)辅助的晶格图案已用于SIM中的结构照明。但是,使用传统的DOE设计方法提高衍射晶格图案的空间频率仍然具有挑战性,因此超分辨率成像性能受到限制。在本文中,我们提出了一种新颖的DOE设计方法,以生成空间频率接近截止频率的晶格图案。这是首次通过衍射光学获得具有如此高空间频率的晶格图案。最后,所提出的SIM在519nm荧光下实现了131nm的横向分辨率,同时通过仅在照明光路中插入单个精心设计的DOE,保持了作为标准倒置荧光显微镜的原始尺寸,这可能会促进这种紧凑型SIM在超分辨率成像领域的应用。