Super-resolution imaging of fast morphological dynamics of neurons in behaving animals

Yujie Zhang; Lu Bai; Xin Wang; Yuchen Zhao; Tianlei Zhang; Lichen Ye; Xufei Du; Zhe Zhang; Jiulin Du; Kai Wang

doi:10.1038/s41592-024-02535-9

Super-resolution imaging of fast morphological dynamics of neurons in behaving animals

Nat Methods. 2024 Nov 22. doi: 10.1038/s41592-024-02535-9. Online ahead of print.

Authors

Yujie Zhang^{1

2}, Lu Bai^{1

2}, Xin Wang^{1

2}, Yuchen Zhao^{1

2}, Tianlei Zhang^{1

2}, Lichen Ye¹, Xufei Du^{1

2}, Zhe Zhang¹, Jiulin Du^{1

2

3}, Kai Wang^{4

5

6}

Affiliations

¹ Institute of Neuroscience, Key Laboratory of Brain Cognition and Brain-inspired Intelligence Technology, CAS Center for Excellence in Brain Science and Intelligence Technology, Chinese Academy of Sciences, Shanghai, China.
² University of Chinese Academy of Sciences, Beijing, China.
³ School of Life Science and Technology, ShanghaiTech University, Shanghai, China.
⁴ Institute of Neuroscience, Key Laboratory of Brain Cognition and Brain-inspired Intelligence Technology, CAS Center for Excellence in Brain Science and Intelligence Technology, Chinese Academy of Sciences, Shanghai, China. wangkai@ion.ac.cn.
⁵ University of Chinese Academy of Sciences, Beijing, China. wangkai@ion.ac.cn.
⁶ School of Life Science and Technology, ShanghaiTech University, Shanghai, China. wangkai@ion.ac.cn.

PMID: 39578627
DOI: 10.1038/s41592-024-02535-9

Abstract

Neurons are best studied in their native states in which their functional and morphological dynamics support animals' natural behaviors. Super-resolution microscopy can potentially reveal these dynamics in higher details but has been challenging in behaving animals due to severe motion artifacts. Here we report multiplexed, line-scanning, structured illumination microscopy, which can tolerate motion of up to 50 μm s^-1 while achieving 150-nm and 100-nm lateral resolutions in its linear and nonlinear forms, respectively. We continuously imaged the dynamics of spinules in dendritic spines and axonal boutons volumetrically over thousands of frames and tens of minutes in head-fixed mouse brains during sleep-wake cycles. Super-resolution imaging of axonal boutons revealed spinule dynamics on a scale of seconds. Simultaneous two-color imaging further enabled analyses of the spatial distributions of diverse PSD-95 clusters and opened up possibilities to study their correlations with the structural dynamics of dendrites in the brains of head-fixed awake mice.