College of Physics Science and Technology, Guangxi Normal University, Guilin, Guangxi, China.
Sino-German College of Intelligent Manufacturing, Shenzhen Technology University, Shenzhen, Guang-dong, China.
J Biophotonics. 2024 Oct;17(10):e202400154. doi: 10.1002/jbio.202400154. Epub 2024 Aug 4.
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.
在这项研究中，我们引入了一种新型的双激光多色成像系统。该系统与多通道滤光轮集成，可以比较三种光谱解卷积算法（非负矩阵分解 [NMF]、RCAN 和 PICASSO），展示了仅使用两个激光源实现四色成像的效果。结合可靠的图像重建算法，四个通道的超分辨率四色图像的空间分辨率分别达到 130、125、133 和 132nm。我们对来自小鼠肝细胞（AML12）、人神经母细胞瘤细胞（SH-SY5Y）、小鼠海马神经元细胞（HT-22）和永生化的骨髓源性巨噬细胞的脂滴、线粒体、溶酶体和核进行了成像。同时，我们观察到 HT-22 和 SH-SY5Y 细胞在遭受氧化应激时发生的染色质浓缩、核收缩、DNA 片段化、凋亡小体形成以及线粒体自噬中 Mito 和 Lyso 的融合。我们的多色 SIM 成像系统为动态细胞器研究和活细胞中的其他高分辨率研究建立了一个强大的平台。
