Open source core technology built into the projector hardware enables high-speed, automatic polarization modulated 3D structured illumination microscopy (SIM) imaging. 3DSIM reconstruction of plant and animal tissue samples: (a) cell walls of oleander leaves, (b) hollow structures in black algae leaves, (c) root tips of corn panicles, and (d) actin filaments in mouse kidney tissues; The corresponding maximum intensity projection (MIP) images are shown in the bottom row (eh) respectively. Scale bar: 2 μm. Credit: Advanced Photonics Nexus (2023). DOI: 10.1117/1.APN.3.1.016001
In the ever-evolving field of microscopy, recent years have seen remarkable advances in both hardware and algorithms, propelling our ability to explore the infinitesimal wonders of life. However, the journey toward three-dimensional structured illumination microscopy (3DSIM) has been hampered by challenges related to the speed and complexity of polarization modulation.
Enter the 3DSIM high-speed modulation system “DMD-3DSIM”, combining a digital display with super-resolution imaging, allowing scientists to see cellular structures in unprecedented detail.
As shown in Advanced Photonics Nexus, the team of Professor Peng Xi from Peking University developed this innovative configuration around a digital micromirror device (DMD) and an electro-optical modulator (EOM). It addresses resolution challenges by significantly improving lateral (side-to-side) and axial (top-to-bottom) resolution, for 3D spatial resolution that is reportedly twice that achieved by traditional wide-field imaging techniques.
In practical terms, this means that DMD-3DSIM can capture intricate details of subcellular structures, such as the nuclear pore complex, microtubules, actin filaments and mitochondria of animal cells. The application of the system has been extended to the study of highly scattering plant cell ultrastructures, such as the cell walls of oleander leaves and the hollow structures of black algae leaves. Even in a mouse kidney slice, the system revealed a pronounced polarization effect in actin filaments.
An open gateway to discovery
What makes DMD-3DSIM even more exciting is its commitment to open science. Xi’s team has made all hardware components and control mechanisms freely available on GitHub, fostering collaboration and encouraging the scientific community to build on this technology.
The DMD-3DSIM technique not only facilitates significant biological discoveries, but also lays the foundation for the next generation of 3DSIM. In applications involving live cell imaging, advances in brighter, more photostable dyes, denoising algorithms, and neural network-based deep learning models promise to improve imaging time, information recovery and real-time restoration of 3DSIM images from noisy data. By combining hardware and software openness, researchers hope to pave the way for the future of multidimensional imaging.
More information:
Yaning Li et al, High-speed self-polarization timing modulation three-dimensional structured illumination microscopy, Advanced Photonics Nexus (2023). DOI: 10.1117/1.APN.3.1.016001
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