Various biological behaviors can only be observed in 3D at high speed over the long term with low phototoxicity. Light-field microscopy (LFM) provides an elegant compact solution to record 3D information in a tomographic manner simultaneously, which can facilitate high photon efficiency. However, LFM still suffers from the missing-cone problem, leading to degraded axial resolution and ringing effects after deconvolution. Here, we propose a mirror-enhanced scanning LFM (MiSLFM) to achieve long-term high-speed 3D imaging at super-resolved axial resolution with a single objective, by fully exploiting the extended depth of field of LFM with a tilted mirror placed below samples. To establish the unique capabilities of MiSLFM, we performed extensive experiments, we observed various organelle interactions and intercellular interactions in different types of photosensitive cells under extremely low light conditions. Moreover, we demonstrated that superior axial resolution facilitates more robust blood cell tracking in zebrafish larvae at high speed.

各种生物行为只能在 3D 中以低光毒性长期高速观察。光场显微镜 (LFM) 提供了一种优雅紧凑的解决方案，可以同时以断层扫描方式记录 3D 信息，从而提高光子效率。然而，LFM 仍然存在缺失锥体问题，导致解卷积后轴向分辨率和振铃效应降低。在这里，我们提出了一种镜面增强扫描 LFM (MiSLFM)，通过在下方放置倾斜镜充分利用 LFM 的扩展景深，以超分辨率轴向分辨率实现长期高速 3D 成像，使用单个物镜样品。为了建立 MiSLFM 的独特功能，我们进行了大量实验，我们在极低光照条件下观察到不同类型光敏细胞的各种细胞器相互作用和细胞间相互作用。此外，我们证明了卓越的轴向分辨率有助于在斑马鱼幼虫中高速追踪更强大的血细胞。