Two-photon synthetic aperture microscopy for minimally invasive fast 3D imaging of native subcellular behaviors in deep tissue

• DOI: 10.1016/j.cell.2023.04.016
• Publication Date: 2023-05-12T14:29:47Z
• Authors: Zhifeng Zhao; Yiliang Zhou; Bo Liu; Jing He; Jiayin Zhao; Yeyi Cai; Jingtao Fan; Xinyang Li; Zilin Wang; Zhi Lu; Jiamin Wu; Hai Qi; Qionghai Dai
• Keywords: Photobleaching; Two-photon excitation microscopy; Autofluorescence; Optical sectioning; Live cell imaging

Holistic understanding of physio-pathological processes requires noninvasive 3D imaging in deep tissue across multiple spatial and temporal scales to link diverse transient subcellular behaviors with long-term physiogenesis. Despite broad applications two-photon microscopy (TPM), there remains an inevitable tradeoff among spatiotemporal resolution, volumes, durations due the point-scanning scheme, accumulated phototoxicity, optical aberrations. Here, we harnessed concept synthetic aperture radar TPM achieve aberration-corrected dynamics at a millisecond scale for over 100,000 large volumes tissue, three orders magnitude reduction photobleaching. With its advantages, identified direct intercellular communications through migrasome generation following traumatic brain injury, visualized formation process germinal center mouse lymph node, characterized heterogeneous cellular states visual cortex, opening up horizon intravital understand organizations functions biological systems holistic level.