Imaging volumetric dynamics at high speed in mouse and zebrafish brain with confocal light field microscopy

• Published in: Nature biotechnology Vol. 39; no. 1; pp. 74 - 83
• Main Authors: Zhang, Zhenkun, Bai, Lu, Cong, Lin, Yu, Peng, Zhang, Tianlei, Shi, Wanzhuo, Li, Funing, Du, Jiulin, Wang, Kai
• Format: Journal Article
• Language: English
• Published: New York Nature Publishing Group US 01.01.2021; Nature Publishing Group
• Subjects: 631/1647/245/2225; 631/1647/245/2226; 631/1647/328/2235; Agriculture; Animals; Bioinformatics; Biomedical and Life Sciences; Biomedical Engineering/Biotechnology; Biomedicine; Biotechnology; Blood cells; Brain; Brain - blood supply; Brain - cytology; Brain - diagnostic imaging; Brain - physiology; Calcium; Calcium - metabolism; Cell Tracking; Danio rerio; Diameters; Fluorescence; Image resolution; Imaging, Three-Dimensional - methods; Larva - cytology; Larva - metabolism; Larva - physiology; Larvae; Life Sciences; Male; Methods; Mice; Mice, Inbred C57BL; Micrometers; Microscopes; Microscopy; Microscopy, Confocal - methods; Neural circuitry; Neural networks; Neuroimaging; Neurons - cytology; Optical sectioning; Prey; Sectioning; Swimming; Vascular system; Zebrafish; China
• ISSN: 1087-0156; 1546-1696
• DOI: 10.1038/s41587-020-0628-7

A detailed understanding of the function of neural networks and how they are supported by a dynamic vascular system requires fast three-dimensional imaging in thick tissues. Here we present confocal light field microscopy, a method that enables fast volumetric imaging in the brain at depths of hundreds of micrometers. It uses a generalized confocal detection scheme that selectively collects fluorescent signals from the in-focus volume and provides optical sectioning capability to improve imaging resolution and sensitivity in thick tissues. We demonstrate recording of whole-brain calcium transients in freely swimming zebrafish larvae and observe behaviorally correlated activities in single neurons during prey capture. Furthermore, in the mouse brain, we detect neural activities at depths of up to 370 μm and track blood cells at 70 Hz over a volume of diameter 800 μm × thickness 150 μm and depth of up to 600 μm. A new imaging method enables deep imaging of neural networks.