ライブセル超解像イメージングのための新顕微鏡技術を開発(Xi Peng Lab Develops Triangle Structured Illumination Microscopy for Sustained Live-Cell Super-Resolution Imaging)

2025-08-18 北京大学(PKU)

北京大学・彭熙教授の研究チームは、新しい三角構造化照明顕微鏡(3I-SIM)を開発し、生細胞の長時間超解像イメージングを実現しました。従来のSIMは3方向の1次元ストライプ照明が必要で撮影速度や光退色に課題がありましたが、3I-SIMは三角ビーム干渉により2次元六角格子照明を生成。7枚のフレームで再構成可能となり、従来比3倍の高速化と光退色低減を達成しました。その結果、最大1,697 Hzでの撮像と13時間以上の連続観察(10万フレーム超)が可能となり、神経成長円錐や小胞体の動態観察などこれまで困難だった生物学的現象を捉えました。この技術は既存2D-SIM装置へのアップグレードも容易で、幅広い研究応用が期待されます。成果はNature Photonicsに掲載されました。

ライブセル超解像イメージングのための新顕微鏡技術を開発(Xi Peng Lab Develops Triangle Structured Illumination Microscopy for Sustained Live-Cell Super-Resolution Imaging)
Figure 1. Principle and performance characterization of 3I-SIM. a. Schematic of the 3I-SIM imaging system. b. Comparison of conventional two-beam interference modulation and 3I-SIM triangular-beam interference modulation patterns. c. Dual-color imaging of synaptonemal complexes, comparing widefield and super-resolution results. d. Imaging of Nile Red–labeled COS-7 cells, comparing widefield and super-resolution results.

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三角ビーム干渉構造照明顕微鏡法 Triangle-beam interference structured illumination microscopy

Yunzhe Fu,Yiwei Hou,Qianxi Liang,Wenyi Wang,Xin Chen,Boya Jin,Jing Ling,Qiuchen Gu,Donghyun Kim,Pengli Zheng,Meiqi Li & Peng Xi
Nature Photonics  Published:15 August 2025
DOI:https://doi.org/10.1038/s41566-025-01730-0

Abstract

Structured illumination microscopy (SIM) is a powerful tool for live-cell super-resolution imaging. Conventional two-dimensional (2D)-SIM uses one-dimensional stripe patterns and rotates them at three angles to achieve uniform resolution. Here, to alleviate photobleaching and improve the temporal resolution of 2D-SIM, we develop triangle-beam interference SIM (3I-SIM), which generates a 2D lattice pattern based on radially polarized beam interference. The radial polarization enhances the signal-to-noise ratio of the high-frequency components. Compared with conventional 2D-SIM, 3I-SIM reduces photobleaching and improves the temporal resolution to 242 Hz. Benefiting from unidirectional phase shift, 3I-SIM provides threefold higher rolling frame rate than conventional 2D-SIM to visualize fast biological dynamics. We further developed 3I-Net, a deep neural network with a co-supervised training scheme, to enhance the performance of 3I-SIM under an extremely low signal intensity. Its higher sensitivity enables the consecutive acquisition of over 100,000 time points at a spatial resolution of 100 nm. We continuously monitor the fine morphological changes in neuronal growth cones for up to 13 h, as well as the transient signals from actin filaments regulating endoplasmic reticulum dynamics. We believe 3I-SIM will offer a suitable platform to study complex and rapid biological processes with high data throughput.

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