In recent years, notable progress has been achieved in both the hardware and algorithms of structured illumination microscopy (SIM). Nevertheless, the advancement of three-dimensional structured illumination microscopy (3DSIM) has been impeded by challenges arising from the speed and intricacy of polarization modulation. We introduce a high-speed modulation 3DSIM system, leveraging the polarization-maintaining and modulation capabilities of a digital micromirror device (DMD) in conjunction with an electro-optic modulator. The DMD-3DSIM system yields a twofold enhancement in both lateral (133 nm) and axial (300 nm) resolution compared to wide-field imaging and can acquire a data set comprising 29 sections of 1024 pixels×1024 pixels, with 15 ms exposure time and 6.75 s per volume. The versatility of the DMD-3DSIM approach was exemplified through the imaging of various specimens, including fluorescent beads, nuclear pores, microtubules, actin filaments, and mitochondria within cells, as well as plant and animal tissues. Notably, polarized 3DSIM elucidated the orientation of actin filaments. Furthermore, the implementation of diverse deconvolution algorithms further enhances 3D resolution. The DMD-based 3DSIM system presents a rapid and reliable methodology for investigating biomedical phenomena, boasting capabilities encompassing 3D superresolution, fast temporal resolution, and polarization imaging.
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 pattern. Opto-electronic devices, such as Spatial Light Modulator (SLM) or Digital Micro-mirror Device (DMD), can provide rapid switch of illumination patterns for SIM. Although DMD is much more cost-effective than SLM, it was previously restricted in association with incoherent light sources. To extend its application with coherent illumination, we model the DMD as a blazed grating, and simulate the effect with DMD pattern changes in SIM. Based on the simulation, we report a fast, high-resolution and cost-efficient SIM with DMD. Our home-built laser interference-based DMD-SIM (LiDMD-SIM) reveals the nuclear pore complex and microtubule in mammalian cells with doubled spatial resolution.
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