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多色结构照明显微镜及利用数字微镜器件对多色光进行定量控制

Multicolor structured illumination microscopy and quantitative control of polychromatic light with a digital micromirror device.

作者信息

Brown Peter T, Kruithoff Rory, Seedorf Gregory J, Shepherd Douglas P

机构信息

Department of Physics and Center for Biological Physics, Arizona State University, Tempe, AZ 85287, USA.

Department of Pediatrics and Pediatric Heart Lung Center, University of Colorado Anschutz Medical Campus, Aurora, CO 80045, USA.

出版信息

Biomed Opt Express. 2021 Jun 1;12(6):3700-3716. doi: 10.1364/BOE.422703.

Abstract

Linear structured illumination microscopy (SIM) is a super-resolution microscopy technique that does not impose photophysics requirements on fluorescent samples. Multicolor SIM implementations typically rely on liquid crystal on silicon (LCoS) spatial light modulators (SLM's) for patterning the excitation light, but digital micromirror devices (DMD's) are a promising alternative, owing to their lower cost and higher speed. However, existing coherent DMD SIM implementations use only a single wavelength of light, limited by the lack of efficient approaches for solving the blazed grating effect for polychromatic light. We develop the requisite quantitative tools, including a closed form solution of the blaze and diffraction conditions, forward models of DMD diffraction and pattern projection, and a model of DMD aberrations. Based on these advances, we constructed a three-color DMD microscope, quantified the effect of aberrations from the DMD, developed a high-resolution optical transfer function measurement technique, and demonstrated SIM on fixed and live cells. This opens the door to applying DMD's in polychromatic applications previously restricted to LCoS SLM's.

摘要

线性结构光照显微镜(SIM)是一种超分辨率显微镜技术,对荧光样品没有光物理要求。多色SIM实现通常依赖于硅基液晶(LCoS)空间光调制器(SLM)来对激发光进行图案化,但数字微镜器件(DMD)因其成本较低和速度较高而成为一种有前途的替代方案。然而,现有的相干DMD SIM实现仅使用单一波长的光,这受到缺乏解决多色光闪耀光栅效应的有效方法的限制。我们开发了必要的定量工具,包括闪耀和衍射条件的闭式解、DMD衍射和图案投影的正向模型以及DMD像差模型。基于这些进展,我们构建了一台三色DMD显微镜,量化了DMD像差的影响,开发了一种高分辨率光学传递函数测量技术,并在固定细胞和活细胞上展示了SIM。这为将DMD应用于以前仅限于LCoS SLM的多色应用打开了大门。

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