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

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.

Data availability

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

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.

Correspondence to Lingjie Kong or Zhenrong Zheng or Qionghai Dai.

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The authors declare no competing interests.

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Supplementary information

41566_2019_474_MOESM2_ESM.mp4

Calcium imaging of cardiac cells with periodic behaviour.

41566_2019_474_MOESM3_ESM.mp4

Calcium imaging of cardiac cells with non-periodic dynamics.

41566_2019_474_MOESM4_ESM.mp4

Calcium imaging of neurons.

41566_2019_474_MOESM5_ESM.mp4

Calcium imaging of human cortical slices.

41566_2019_474_MOESM6_ESM.mp4

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

41566_2019_474_MOESM7_ESM.mp4

Tracking of immune cells.

41566_2019_474_MOESM8_ESM.mp4

Imaging of vascular and calcium signals in awake mice.

41566_2019_474_MOESM9_ESM.mp4

Calcium imaging of neural activity in mice.

41566_2019_474_MOESM10_ESM.mp4

Calcium imaging of cardiac tissue in a microfluidic device.

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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