Structured illumination microscopy using digital micro-mirror device and coherent light source
Structured illumination microscopy using digital micro-mirror device and coherent light source
复制标题
使用数字微镜器件和相干光源的结构照明显微镜
DOI：
10.1063/5.0008264
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发表时间：
2020-06-08
影响因子：
4
通讯作者：
Xi, Peng
中科院分区：
文献类型：
--
作者：
Li, Meiqi;Li, Yaning;Xi, Peng
结构照明显微镜（SIM）实现双倍的空间分辨率，通过激发试样与高对比度，高频正弦模式。这种照明图案可以由激光干涉或非相干结构图案产生。光电器件，如空间光调制器（SLM）或数字微镜器件（DMD），可以为SIM提供快速的照明模式切换。虽然DMD比SLM更具成本效益，但由于其衍射阶数与入射角和相干入射波长有关，因此它以前受到非相干光源的限制。为了扩展其在相干照明中的应用，本文将DMD建模为燃烧光栅，并在SIM卡中模拟了DMD模式变化的效果。通过仔细分析不同角度和相位的照明对比度，我们报告了一种具有DMD调制的快速，高分辨率和经济高效的SIM。我们自制的基于激光干涉的DMD-SIM （LiDMD-SIM）以双倍的空间分辨率揭示了哺乳动物细胞的核孔复合物和微管。我们进一步提出了多色LiDMD-SIM概念，联合使用不同波长不同入射角的DMD开/关状态，具有高对比度和最大分辨率增强。
Structured illumination microscopy (SIM) achieves doubled spatial resolution through exciting the specimen with high-contrast, high-frequency sinusoidal patterns. Such an illumination pattern can be generated by laser interference or incoherent structured patterns. Opto-electronic devices, such as a Spatial Light Modulator (SLM) or a Digital Micro-mirror Device (DMD), can provide rapid switch of illumination patterns for SIM. Although the DMD is much more cost-effective than the SLM, it was previously restricted in association with incoherent light sources, as its diffractive orders are related to the incident angle and the wavelength of coherent incidence. To extend its application with coherent illumination, here, we model the DMD as a blazed grating and simulate the effect with DMD pattern changes in the SIM. With careful analysis of the illumination contrast along different angles and phases, we report a fast, high-resolution, and cost-efficient SIM with DMD modulation. Our home-built laser interference-based DMD-SIM (LiDMD-SIM) reveals the nuclear pore complex and microtubule in mammalian cells with doubled spatial resolution. We further proposed the multi-color LiDMD-SIM concept by jointly employing the DMD ON/OFF states with different incident angles for different wavelengths, with high contrast and maximum resolution enhancement.