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Iterative tomography with digital adaptive optics permits hou
LV-GCamp6s was used to infect the human 3D cerebral organoids. (From BrainVTA)
The viruses used in this article are in the table below
Custom made LVs  rLV-EF1a-GCamp6s-WPRE
Jiamin Wu, Zhi Lu, Dong Jiang, Yuduo Guo, Hui Qiao, Yi Zhang, Tianyi Zhu, Yeyi Cai, Xu Zhang, Karl Zhanghao , Hao Xie, Tao Yan, Guoxun Zhang, Xiaoxu Li, Zheng Jiang, Xing Lin, Lu Fang, Bing Zhou, Peng Xi, Jingtao Fan, Li Yu, Qionghai Dai
Pub Date: 2021-05-25, DOI: 10.1016/j.cell.2021.04.029, Email: [email protected]
Long-term subcellular intravital imaging in mammals is vital to study diverse intercellular behaviors and organelle functions during native physiological processes. However, optical heterogeneity, tissue opacity, and phototoxicity pose great challenges. Here, we propose a computational imaging framework, termed digital adaptive optics scanning light-field mutual iterative tomography (DAOSLIMIT), featuring high-speed, high-resolution 3D imaging, tiled wavefront correction, and low phototoxicity with a compact system. By tomographic imaging of the entire volume simultaneously, we obtained volumetric imaging across 225 × 225 × 16 μm3, with a resolution of up to 220 nm laterally and 400 nm axially, at the millisecond scale, over hundreds of thousands of time points. To establish the capabilities, we investigated large-scale cell migration and neural activities in different species and observed various subcellular dynamics in mammals during neutrophil migration and tumor cell circulation.

Figure 1. 3D calcium dynamics in human cerebral organoids, Drosophila larvae, and zebrafish larvae.
In this study, ultrahigh-resolution fluorescence imaging with low phototoxicity enables 3D visualization of fast cellular and subcellular processes in organoids, zebrafish, and mammals.
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