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<p>Despite its proven value for biomedical research, super-resolution structured illumination microscopy still faces challenges in both fidelity of image reconstruction and imaging speed. Substantial background interference introduces artefacts and degrades resolution, while computationally intensive image reconstruction and illumination pattern switching limit imaging throughput. Here we present digital array modulation microscopy (DaMo), which combines digital array modulation with a single-spectrum reconstruction algorithm. Gaussian illumination modulation combined with digital detection modulation enables heterodyne detection with a 100% contrast. Therefore, DaMo achieves high-fidelity reconstruction (Pearson correlation coefficient 0.99 ± 0.01) under substantial background interference, with a 102× faster reconstruction speed than state-of-the-art super-resolution structured illumination microscopy processing. DaMo offers an axial resolution of 300 nm and a lateral resolution of 100 nm while achieving a 1,284-fold improvement in the signal-to-background ratio in whole-cell three-dimensional imaging. DaMo operates without additional modulators or extra image enhancement, providing artefact-free precision with a streamlined workflow. We demonstrate the versatility of DaMo via quantitative live-cell imaging of actin dynamics, tracking of cascaded filopodia fusion events, multicolour whole-smear imaging for cell cycle profiling, and tissue pathology assessment of intestinal epithelial injury in mitochondria. DaMo paves the way for large-scale, background-suppressed super-resolution imaging across diverse biological systems.</p>

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Three-dimensional super-resolution imaging with suppressed background via digital array modulation microscopy

  • 期刊论文

  • 发表日期：

    2026年3月26日

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  • Sijie Li,
  • Rui Jin,
  • Mengqi Lu,
  • Ning Zhang,
  • Meng Zhang,
  • Yunfei Wei,
  • Zhan Zhang,
  • Shirui Guo,
  • Yu-Hui Zhang,
  • Fengchao Wang,
  • Hui Gong,
  • Qingming Luo,
  • Jing Yuan

摘要

Despite its proven value for biomedical research, super-resolution structured illumination microscopy still faces challenges in both fidelity of image reconstruction and imaging speed. Substantial background interference introduces artefacts and degrades resolution, while computationally intensive image reconstruction and illumination pattern switching limit imaging throughput. Here we present digital array modulation microscopy (DaMo), which combines digital array modulation with a single-spectrum reconstruction algorithm. Gaussian illumination modulation combined with digital detection modulation enables heterodyne detection with a 100% contrast. Therefore, DaMo achieves high-fidelity reconstruction (Pearson correlation coefficient 0.99 ± 0.01) under substantial background interference, with a 102× faster reconstruction speed than state-of-the-art super-resolution structured illumination microscopy processing. DaMo offers an axial resolution of 300 nm and a lateral resolution of 100 nm while achieving a 1,284-fold improvement in the signal-to-background ratio in whole-cell three-dimensional imaging. DaMo operates without additional modulators or extra image enhancement, providing artefact-free precision with a streamlined workflow. We demonstrate the versatility of DaMo via quantitative live-cell imaging of actin dynamics, tracking of cascaded filopodia fusion events, multicolour whole-smear imaging for cell cycle profiling, and tissue pathology assessment of intestinal epithelial injury in mitochondria. DaMo paves the way for large-scale, background-suppressed super-resolution imaging across diverse biological systems.