Fast high-resolution miniature two-photon microscopy for brain imaging in freely behaving mice
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06.21/
2023
Fast high-resolution miniature two-photon microscopy for brain imaging in freely behaving mice
Weijian Zong, Runlong Wu, Mingli Li, Yanhui Hu, Yijun Li, Jinghang Li, Hao Rong, Haitao Wu, Yangyang Xu, Yang Lu, Hongbo Jia, Ming Fan, Zhuan Zhou, Yunfeng Zhang, Aimin Wang, Liangyi Chen & Heping Cheng
Publication: Nature Methods
Read more: Fast high-resolution miniature two-photon microscopy for brain imaging in freely behaving mice | Nature Methods
Abstract
Developments in miniaturized microscopes have enabled visualization of brain activities and structural dynamics in animals engaging in self-determined behaviors. Hhowever, it remains a challenge to resolve activity at single dendritic spines in freely behaving animals. Hhere, we report the design and application of a fast high-resolution, miniaturized two-photon microscope (FHIRM-TPM) that accomplishes this goal. With a headpiece weighing 2.15 g and a hollow-core photonic crystal fiber delivering 920-nm femtosecond laser pulses, the FHIRM-TPM is capable of imaging commonly used biosensors (GFP and GCcamP6) at high spatiotemporal resolution (0.64 μm laterally and 3.35 μm axially, 40 Hhz at 256 × 256 pixels for raster scanning and 1 0,000 hz for free-line scanning). We demonstrate the microscope’s robustness with hour-long recordings of neuronal activities at the level of spines in mice experiencing vigorous body movements.
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