Multi-color two-laser super-resolution structured illumination microscopy for the visualization of multi-organelle in living cells

IF 2 3区 物理与天体物理 Q3 BIOCHEMICAL RESEARCH METHODS
Xuejuan Hu, Yadan Tan, Yujie Huang, Jianze Ye, Yifei Liang, Xiaokun Yang, Hengliang Wang, Zihao Cheng, Lihu Wang, Shiqian Liu, Minfei Li, Zhengdi He, Qianding Gao, Jingli Zhong
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Abstract

In this study, we introduced a novel dual-laser multi-color imaging system. Integrated with a multi-channel filter wheel, this system compared three spectral decontamination algorithms (nonnegative matrix factorization [NMF], RCAN, and PICASSO) showcasing its efficacy in achieving four-color imaging with only two laser sources. Combined with a reliable image reconstruction algorithm, the spatial resolution of four channels super-resolution four-color images reached 130, 125, 133, and 132 nm, respectively. Lipid droplets, mitochondria, lysosomes, and nuclei from the mouse hepatocytes (AML12), human neuroblastoma cells (SH-SY5Y), mouse hippocampal neuronal cells (HT-22), and immortalized murine bone marrow-derived macrophages were imaged. At the same time, the chromatin condensation, nuclear contraction, DNA fragmentation, apoptotic body formation, as well as the fusion of Mito and Lyso involved in mitochondrial autophagy were observed in HT-22 and SH-SY5Y cells suffering oxidative stress. Our multi-color SIM imaging system establishes a powerful platform for dynamic organelle studies and other high-resolution investigations in live cells.

Abstract Image

用于活细胞多细胞器可视化的多色双激光超分辨率结构照明显微镜。
在这项研究中,我们介绍了一种新型双激光多色成像系统。该系统集成了一个多通道滤光轮,比较了三种光谱净化算法(非负矩阵因式分解[NMF]、RCAN 和 PICASSO),展示了其在仅使用两个激光源实现四色成像方面的功效。结合可靠的图像重建算法,四通道超分辨率四色图像的空间分辨率分别达到了 130、125、133 和 132 nm。对小鼠肝细胞(AML12)、人神经母细胞瘤细胞(SH-SY5Y)、小鼠海马神经元细胞(HT-22)和永生化小鼠骨髓巨噬细胞的脂滴、线粒体、溶酶体和细胞核进行了成像。同时,在遭受氧化应激的 HT-22 和 SH-SY5Y 细胞中观察到染色质凝结、核收缩、DNA 断裂、凋亡体形成以及参与线粒体自噬的 Mito 和 Lyso 融合。我们的多色 SIM 成像系统为活细胞中的动态细胞器研究和其他高分辨率研究建立了一个强大的平台。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Journal of Biophotonics
Journal of Biophotonics 生物-生化研究方法
CiteScore
5.70
自引率
7.10%
发文量
248
审稿时长
1 months
期刊介绍: The first international journal dedicated to publishing reviews and original articles from this exciting field, the Journal of Biophotonics covers the broad range of research on interactions between light and biological material. The journal offers a platform where the physicist communicates with the biologist and where the clinical practitioner learns about the latest tools for the diagnosis of diseases. As such, the journal is highly interdisciplinary, publishing cutting edge research in the fields of life sciences, medicine, physics, chemistry, and engineering. The coverage extends from fundamental research to specific developments, while also including the latest applications.
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