Kilohertz volumetric imaging of in vivo dynamics using squeezed light field microscopy

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Zhaoqiang Wang, Ruixuan Zhao, Daniel A Wagenaar, Diego Espino, Liron Sheintuch, Ohr Benshlomo, Wenjun Kang, Enbo Zhu, Calvin K Lee, William C Schmidt, Aryan Pammar, Jing Wang, Gerard C L Wong, Rongguang Liang, Sungmoo Lee, Michael Z Lin, Madhuvanthi Kannan, Peyman Golshani, Tzung K Hsiai, Liang Gao

Nat Methods. 2025 Oct;22(10):2194-2204. doi: 10.1038/s41592-025-02843-8. Epub 2025 Sep 23.

ABSTRACT

Volumetric functional imaging of transient cellular signaling and motion dynamics is often limited by hardware bandwidth and the scarcity of photons under short exposures. To overcome these challenges, we introduce squeezed light field microscopy (SLIM), a computational imaging approach that rapidly captures high-resolution three-dimensional light signals using only a single, low-format camera sensor. SLIM records over 1,000 volumes per second across a 550-µm diameter field of view and 300-µm depth, achieving 3.6-µm lateral and 6-µm axial resolution. Here we demonstrate its utility in blood cell velocimetry within the embryonic zebrafish brain and in freely moving tails undergoing high-frequency swings. Millisecond-scale temporal resolution further enables precise voltage imaging of neural membrane potentials in the leech ganglion and hippocampus of behaving mice. Together, these results establish SLIM as a versatile and robust tool for high-speed volumetric microscopy across diverse biological systems.

PMID:40987870 | DOI:10.1038/s41592-025-02843-8