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Video-rate imaging of biological dynamics at centimetre scale and micrometre resolution

Nature Photonics volume 13, pages 809–816 (2019)Cite this article

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Abstract

Large-scale imaging of biological dynamics with high spatiotemporal resolution is indispensable to system biology studies. However, conventional microscopes have an inherent compromise between the achievable field of view and spatial resolution due to the space–bandwidth product theorem. In addition, a further challenge is the ability to handle the enormous amount of data generated by a large-scale imaging platform. Here, we break these bottlenecks by proposing the use of a flat–curved–flat imaging strategy, in which the sample plane is magnified onto a large spherical image surface and then seamlessly conjugated to multiple planar sensors. Our real-time, ultra-large-scale, high-resolution (RUSH) imaging platform operates with a 10 × 12 mm2 field of view, a uniform resolution of ~1.20 μm after deconvolution and a data throughput of 5.1 gigapixels per second. We use the RUSH platform to perform video-rate, gigapixel imaging of biological dynamics at centimetre scale and micrometre resolution, including brain-wide structural imaging and functional imaging in awake, behaving mice.

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Fig. 1: Schematic and characterization of a RUSH system.
Fig. 2: High-throughput calcium imaging of cardiac cellular ensembles and neuron ensembles.
Fig. 3: In vitro calcium imaging of spontaneous epileptiform activity in acute human cortical slices.
Fig. 4: In vivo brain-wide imaging of a Cx3Cr1-GFP mouse.
Fig. 5: Simultaneous imaging of vascular morphological dynamics and neural calcium signals in awake, behaving mice.
Fig. 6: In vivo brain-wide calcium imaging of adult C57BL/6 mice at dendritic resolution.

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The data that support the plots within this paper and other findings of this study are available from the corresponding authors upon reasonable request.

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Acknowledgements

We thank Y. Shu, S. Wang, Y. Li, Z. Guo, B. Zhou, W. Wang and Y. Jia for assistance with sample preparation. We also thank members of Dai’s group for helping with experiments and data analysis: X. Zhang, Y. Zhang, Y. Cheng, K. Liu, C. Zhuang, C. Qiao, Z. Zhao, X. Han, T. Zhou, Y. Zhang, X. Chen, W. Liu, T. Yan, G. Zhang, L. Wang, Y. Ma, X. Hu, J. Hu, T. Zhu, X. Chen, J. Bao and X. Zhang. We acknowledge P. Xi, L. Fang and G. Holtom for their assistance with editing the manuscript. This work is supported by the National Natural Science Foundation of China (61327902, 61671265, 61771287, 61741116, 61722110, 61831014, 31430038 and 81571275) and the Beijing Municipal Science & Technology Commission (Z181100003118014).

Author information

Author notes

  1. These authors contributed equally: Jingtao Fan, Jinli Suo, Jiamin Wu, Hao Xie.

Authors and Affiliations

  1. Department of Automation, Tsinghua University, Beijing, China

    Jingtao Fan, Jinli Suo, Jiamin Wu, Hao Xie, Feng Chen & Qionghai Dai

  2. State Key Laboratory of Modern Optical Instrumentation, Zhejiang University, Hangzhou, China

    Yibing Shen & Zhenrong Zheng

  3. Department of Electronic Engineering, Tsinghua University, Beijing, China

    Guijin Wang

  4. State Key Laboratory of Precision Measurement Technology and Instruments, Department of Precision Instrument, Tsinghua University, Beijing, China

    Liangcai Cao, Guofan Jin & Lingjie Kong

  5. State Key Laboratory of Cognitive Neuroscience and Learning & IDG/McGovern Institute for Brain Research, Beijing Normal University, Beijing, China

    Quansheng He

  6. Department of Neurosurgery, Brain Institute, and Department of Neurology, Epilepsy Centre, Sanbo Brain Hospital, Capital Medical University, Beijing, China

    Tianfu Li & Guoming Luan

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Contributions

J.F., J.S. and J.W. designed the system architecture. J.F. performed system integration and calibration, aided by F.C. and G.W. J.W. and H.X. conducted performance optimization. H.X. conducted most biological experiments, data capturing and analysis, and Y.S. led the optical and mechanical manufacturing. F.C. and G.W. developed the graphical user interface software. L.C. and G.J. contributed to the optimization of system design. Q.H. designed and conducted experiments on acute human cortical slices from epilepsy patients. T.L. and G.L. contributed to human tissue processing. L.K. conceived and supervised the biological experiments and data analysis, aided by H.X. Z.Z. designed the optical system, aided by J.F., J.S. and J.W. Q.D. conceived and supervised the project, and all authors contributed to the writing and editing of the manuscript.

Corresponding authors

Correspondence to Lingjie Kong, Zhenrong Zheng or Qionghai Dai.

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Supplementary information

Supplementary Information

Objective lens design, optical transfer function, point spread function, calcium imaging data.

Supplementary Video 1

Calcium imaging of cardiac cells with periodic behaviour.

Supplementary Video 2

Calcium imaging of cardiac cells with non-periodic dynamics.

Supplementary Video 3

Calcium imaging of neurons.

Supplementary Video 4

Calcium imaging of human cortical slices.

Supplementary Video 5

Calcium imaging of human cortical slices with single-cell resolution.

Supplementary Video 6

Tracking of immune cells.

Supplementary Video 7

Imaging of vascular and calcium signals in awake mice.

Supplementary Video 8

Calcium imaging of neural activity in mice.

Supplementary Video 9

Calcium imaging of cardiac tissue in a microfluidic device.

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Fan, J., Suo, J., Wu, J. et al. Video-rate imaging of biological dynamics at centimetre scale and micrometre resolution. Nat. Photonics 13, 809–816 (2019). https://doi.org/10.1038/s41566-019-0474-7

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