2. Academic Validation
  3. Fast reconstruction and optical-sectioning three-dimensional structured illumination microscopy

Fast reconstruction and optical-sectioning three-dimensional structured illumination microscopy

  • Innovation (Camb). 2025 Jan 12;6(2):100757. doi: 10.1016/j.xinn.2024.100757.
Ruijie Cao 1 2,  Yaning Li 1 2 3,  Wenyi Wang 1 2 4,  Yunzhe Fu 1 2,  Xiaoyu Bu 5,  Dilizhatai Saimi 6,  Jing Sun 5,  Xichuan Ge 4,  Shan Jiang 7,  Yuru Pei 8,  Baoxiang Gao 4,  Zhixing Chen 1 2 9,  Meiqi Li 10,  Peng Xi 1 2
Affiliations

Affiliations

  • 1 Department of Biomedical Engineering, College of Future Technology, Peking University, Beijing 100871, China.
  • 2 National Biomedical Imaging Center, College of Future Technology, Peking University, Beijing 100871, China.
  • 3 China Academy of Space Technology, Beijing Institute of Space Mechanics and Electricity, Beijing 100094, China.
  • 4 Airy Technologies Co., Ltd., Beijing 100081, China.
  • 5 Key Laboratory of Analytical Science and Technology of Hebei Province, College of Chemistry and Materials Science, Hebei University, Baoding 071002, China.
  • 6 College of Future Technology, Institute of Molecular Medicine, National Biomedical Imaging Center, Beijing Key Laboratory of Cardiometabolic Molecular Medicine, Peking University, Beijing 100871, China.
  • 7 Institute of Biomedical Engineering, Beijing Institute of Collaborative Innovation, Beijing, China.
  • 8 Key Laboratory of Machine Perception (MOE), Department of Machine Intelligence, Peking University, Beijing 100871, China.
  • 9 Peking-Tsinghua Center for Life Science, Academy for Advanced Interdisciplinary Studies, Peking University, Beijing 100871, China.
  • 10 School of Life Sciences, Peking University, Beijing 100871, China.
PMID: 39991474 DOI: 10.1016/j.xinn.2024.100757
Abstract

Three-dimensional structured illumination microscopy (3DSIM) is a popular method for observing subcellular/cellular structures or animal/plant tissues with gentle phototoxicity and 3D super-resolution. However, its time-consuming reconstruction process poses challenges for high-throughput imaging and real-time observation. Moreover, traditional 3DSIM typically requires more than six z layers for successful reconstruction and is susceptible to defocused backgrounds. This poses a great gap between single-layer 2DSIM and 6-layer 3DSIM, and limits the observation of thicker samples. To address these limitations, we developed FO-3DSIM, a novel method that integrates spatial-domain reconstruction with optical-sectioning SIM. FO-3DSIM enhances reconstruction speed by up to 855.7 times with superior performance with limited z layers and under high defocused backgrounds. It retains the high-fidelity, low-photon reconstruction capabilities of our previously proposed Open-3DSIM. Utilizing fast reconstruction and optical sectioning, we achieved large field-of-view (FOV) 3D super-resolution imaging of mouse kidney actin, covering a region of 0.453 mm × 0.453 mm × 2.75 μm within 23 min of acquisition and 13 min of reconstruction. Near real-time performance was demonstrated in live actin imaging with FO-3DSIM. Our approach reduces photodamage through limited z layer reconstruction, allowing the observation of ER tubes with just three layers. We anticipate that FO-3DSIM will pave the way for near real-time, large FOV 6D imaging, encompassing xyz super-resolution, multi-color, long-term, and polarization imaging with less photodamage, removed defocused backgrounds, and reduced reconstruction time.

Keywords

large field of view imaging; optical sectioning; real-time observation; reconstruction speed; three-dimensional structure illumination.

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