Super-resolution fluorescence-assisted diffraction computational tomography reveals the threedimensional landscape of the cellular organelle interactome

• 作者: Dashan Dong Xiaoshuai Huang Liuju Li Heng Mao Yanquan Mo Guangyi Zhang Zhe Zhang Jiayu Shen Wei Liu Zeming Wu Guanghui Liu Yanmei Liu Hong Yang Qihuang Gong Kebin Shi Liangyi Chen
• 来源期刊: 光：科学与应用（英文版）
• 年，卷（期）: 2020,（1）
• 所属期刊栏目: Original Articles
• 页码范围: 99-113
• 页数: 15页
• 语种: 英文

摘要

The emergence of super-resolution (SR) fluorescence microscopy has rejuvenated the search for new cellular sub-structures. However, SR fluorescence microscopy achieves high contrast at the expense of a holistic view of the interacting partners and surrounding environment. Thus, we developed SR fluorescence-assisted diffraction computational tomography (SR-FACT), which combines label-free three-dimensional optical diffraction tomography (ODT) with two-dimensional fluorescence Hessian structured illumination microscopy. The ODT module is capable of resolving the mitochondria, lipid droplets, the nuclear membrane, chromosomes, the tubular endoplasmic reticulum, and lysosomes. Using dual-mode correlated live-cell imaging for a prolonged period of time, we observed novel subcellular structures named dark-vacuole bodies, the majority of which originate from densely populated perinuclear regions, and intensively interact with organelles such as the mitochondria and the nuclear membrane before ultimately collapsing into the plasma membrane. This work demonstrates the unique capabilities of SR-FACT, which suggests its wide applicability in cell biology in general.