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.

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In light-sheet fluorescence microscopy, the axial resolution and field of view are mutually constrained. Axially swept light-sheet microscopy (ASLM) can decouple the trade-off, but the confocal detection scheme using a rolling shutter also rejects fluorescence signals from the specimen in the field of interest, which sacrifices the photon efficiency. Here, we report a laterally swept light-sheet microscopy (LSLM) scheme in which the focused beam is first scanned along the axial direction and subsequently laterally swept with the rolling shutter. We show that LSLM can obtain a higher photon efficiency when similar axial resolution and field of view can be achieved. Moreover, based on the principle of image scanning microscopy, applying the pixel reassignment to the LSLM images, hereby named iLSLM, improves the optical sectioning. Both simulation and experimental results demonstrate the higher photon efficiency with similar axial resolution and optical sectioning. Our proposed scheme is suitable for volumetric imaging of specimens that are susceptible to photobleaching or phototoxicity.

The pixel size of a charge-coupled device (CCD) camera plays a major role in the image resolution, and the square pixels are attributed to the physical anisotropy of the sampling frequency. We synthesize the high sampling frequency directions from multiple frames acquired with different angles to enhance the resolution by 1.4 × over conventional CCD orthogonal sampling. To directly demonstrate the improvement of frequency-domain diagonal extension (FDDE) microscopy, lens-free microscopy is used, as its resolution is dominantly determined by the pixel size. We demonstrate the resolution enhancement with a mouse skin histological specimen and a clinical blood smear sample. Further, FDDE is extended to lens-based photography with an ISO 12233 resolution target. This method paves a new way for enhancing the image resolution for a variety of imaging techniques in which the resolution is primarily limited by the sampling pixel size, for example, microscopy, photography, and spectroscopy.