对生理病理过程的整体理解需要在多个空间和时间尺度上对深层组织进行非侵入性3D成像,以将各种瞬时亚细胞行为与长期生理发生联系起来。尽管双光子显微镜(TPM)的广泛应用,时空分辨率之间仍然存在不可避免的权衡,成像体积,以及由于点扫描方案而导致的持续时间,累积的光毒性,和光学像差。这里,我们利用TPM中的合成孔径雷达的概念,在超过100,000个大体积的深层组织中,以毫秒为尺度实现亚细胞动力学的像差校正3D成像,光漂白减少三个数量级。凭借其优势,我们确定了创伤性脑损伤后通过migrasome生成的直接细胞间通讯,可视化小鼠淋巴结生发中心的形成过程,并在小鼠视觉皮层中表征异质细胞状态,为活体成像开辟了视野,从整体上了解生物系统的组织和功能。
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 of two-photon microscopy (TPM), there remains an inevitable tradeoff among spatiotemporal resolution, imaging volumes, and durations due to the point-scanning scheme, accumulated phototoxicity, and optical aberrations. Here, we harnessed the concept of synthetic aperture radar in TPM to achieve aberration-corrected 3D imaging of subcellular dynamics at a millisecond scale for over 100,000 large volumes in deep tissue, with three orders of magnitude reduction in photobleaching. With its advantages, we identified direct intercellular communications through migrasome generation following traumatic brain injury, visualized the formation process of germinal center in the mouse lymph node, and characterized heterogeneous cellular states in the mouse visual cortex, opening up a horizon for intravital imaging to understand the organizations and functions of biological systems at a holistic level.