Zhang Youchang, Asghari Parisa, Scriven David R L, Moore Edwin D W, Chou Keng C
Opt Lett. 2023 Aug 1;48(15):3933-3936. doi: 10.1364/OL.494655.
Among various super-resolution microscopic techniques, structured illumination microscopy (SIM) stands out for live-cell imaging because of its higher imaging speed. However, conventional SIM lacks optical sectioning capability. Here we demonstrate a new, to the best of our knowledge, approach using a phase-modulated spinning disk (PMSD) that enhances the optical sectioning capability of SIM. The PMSD consists of a pinhole array for confocal imaging and a transparent polymer layer for light phase modulation. The light phase modulation was designed to cancel the zeroth-order diffracted beam and create a sharp lattice illumination pattern using the interference of four first-order diffracted beams. In the detection optical path, the PMSD serves as a spatial filter to physically reject about 80% of the out-of-focus signals, an approach that allows for real-time optical reconstruction of super-resolved images with enhanced contrast. Furthermore, the simplicity of the design makes it easy to upgrade a conventional fluorescence microscope to a PMSD SIM system.
在各种超分辨率显微技术中，结构光照明显微镜（SIM）因其较高的成像速度而在活细胞成像方面脱颖而出。然而，传统的SIM缺乏光学切片能力。在此，据我们所知，我们展示了一种使用相位调制旋转盘（PMSD）的新方法，该方法增强了SIM的光学切片能力。PMSD由用于共焦成像的针孔阵列和用于光相位调制的透明聚合物层组成。光相位调制旨在消除零阶衍射光束，并利用四个一阶衍射光束的干涉产生清晰的晶格照明图案。在检测光路中，PMSD用作空间滤波器，物理上滤除约80%的离焦信号，这种方法能够实时光学重建具有增强对比度的超分辨图像。此外，该设计的简单性使得将传统荧光显微镜升级为PMSD SIM系统变得容易。
