近日，北京大学的席鹏与南方科技大学的金大勇等人合作并取得一项新进展。经过不懈努力，他们利用并行采集-读出结构照明显微镜(PAR-SIM)实现了超高时空分辨率成像。相关研究成果已于2024年5月29日在国际知名学术期刊《光：科学与应用》上发表。

在本文中，研究人员提出一种在硬件层面提高SIM帧速率和感兴趣区域(ROI)覆盖的方法，而无需增加相机费用或复杂的算法。在这里，并行采集-读出SIM (PAR-SIM)在当前可用的探测器灵敏度下实现了荧光成像的最高成像速度。通过同步模式生成和图像曝光读出过程，利用探测器的全帧宽度，研究人员实现了惊人的信息时空通量，达到132.9 MPixels·s^-1，是最新技术的9.6倍，最低信噪比为2.11dB，分辨率为100nm。

PAR-SIM在双重激励下展现出了卓越的能力，成功重建了多种细胞器，即便在信号较弱的条件下也表现优异，这得益于其超短的曝光时间。特别值得一提的是，在活的COS-7细胞中，PAR-SIM以惊人的408Hz帧率记录下了线粒体动态管化和正在进行的膜融合过程。研究人员认为，这种新型的并行曝光读出模式不仅大幅提升了SIM模式的调制效率，实现了更高的帧速率，而且通过其战略控制方法，还具有为其他复杂成像系统带来显著效益的潜力。

据悉，结构照明显微镜(SIM)已经成为一种很有前途的超分辨率荧光成像技术，提供不同的配置和计算策略来减轻生物标本实时成像过程中的光毒性。传统的提高系统帧率的努力主要集中在处理算法上，如滚动重建或简化帧重建，或者投资于具有加速行读出速率的昂贵的sCMOS相机。

附：英文原文

Title: Ultra-high spatio-temporal resolution imaging with parallel acquisition-readout structured illumination microscopy (PAR-SIM)

Author: Xu, Xinzhu, Wang, Wenyi, Qiao, Liang, Fu, Yunzhe, Ge, Xichuan, Zhao, Kun, Zhanghao, Karl, Guan, Meiling, Chen, Xin, Li, Meiqi, Jin, Dayong, Xi, Peng

Issue&Volume: 2024-05-29

Abstract: Structured illumination microscopy (SIM) has emerged as a promising super-resolution fluorescence imaging technique, offering diverse configurations and computational strategies to mitigate phototoxicity during real-time imaging of biological specimens. Traditional efforts to enhance system frame rates have concentrated on processing algorithms, like rolling reconstruction or reduced frame reconstruction, or on investments in costly sCMOS cameras with accelerated row readout rates. In this article, we introduce an approach to elevate SIM frame rates and region of interest (ROI) coverage at the hardware level, without necessitating an upsurge in camera expenses or intricate algorithms. Here, parallel acquisition-readout SIM (PAR-SIM) achieves the highest imaging speed for fluorescence imaging at currently available detector sensitivity. By using the full frame-width of the detector through synchronizing the pattern generation and image exposure-readout process, we have achieved a fundamentally stupendous information spatial-temporal flux of 132.9 MPixels · s^-1, 9.6-fold that of the latest techniques, with the lowest SNR of 2.11dB and 100nm resolution. PAR-SIM demonstrates its proficiency in successfully reconstructing diverse cellular organelles in dual excitations, even under conditions of low signal due to ultra-short exposure times. Notably, mitochondrial dynamic tubulation and ongoing membrane fusion processes have been captured in live COS-7 cell, recorded with PAR-SIM at an impressive 408Hz. We posit that this novel parallel exposure-readout mode not only augments SIM pattern modulation for superior frame rates but also holds the potential to benefit other complex imaging systems with a strategic controlling approach.

DOI: 10.1038/s41377-024-01464-8

Source: https://www.nature.com/articles/s41377-024-01464-8