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Large-field-of-view imaging by multi-pupil adaptive optics

Nature Methods volume 14, pages 581583 (2017) | Download Citation

Abstract

Adaptive optics can correct for optical aberrations. We developed multi-pupil adaptive optics (MPAO), which enables simultaneous wavefront correction over a field of view of 450 × 450 μm2 and expands the correction area to nine times that of conventional methods. MPAO's ability to perform spatially independent wavefront control further enables 3D nonplanar imaging. We applied MPAO to in vivo structural and functional imaging in the mouse brain.

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Acknowledgements

This work was funded by NIH grant no. 1U01NS094341-01 and Purdue University. The authors thank W. Gan for valuable discussion and advice; G. Holtom and B. Wei for their help on manuscript preparation; and the Howard Hughes Medical Institute for equipment support. J.-H.P. thanks the NRF (grant no. 2016R1C1B2015130) for support during the manuscript preparation; and L.K. thanks the NSFC (grant no. 61327902) for support during manuscript revision.

Author information

Author notes

    • Jung-Hoon Park
    •  & Lingjie Kong

    Present addresses: J.-H.P. (Department of Biomedical Engineering, Ulsan National Institute of Science and Technology, Ulsan, Republic of Korea) and L.K. (Department of Precision Instrument, Tsinghua University, Beijing, China.

    • Jung-Hoon Park
    •  & Lingjie Kong

    These authors contributed equally to this work.

Affiliations

  1. School of Electrical and Computer Engineering, Purdue University, West Lafayette, Indiana, USA.

    • Jung-Hoon Park
    • , Lingjie Kong
    • , Yifeng Zhou
    •  & Meng Cui
  2. Department of Biological Sciences, Purdue University, West Lafayette, Indiana, USA.

    • Meng Cui
  3. Integrated Imaging Cluster, Purdue University, West Lafayette, Indiana, USA.

    • Meng Cui
  4. Bindley Bioscience Center, Purdue University, West Lafayette, Indiana, USA.

    • Meng Cui

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Contributions

M.C. invented MPAO, developed the experimental schemes of wavefront correction and nonplanar imaging, designed the liquid-immersion-based tunable prism array, and supervised the project. J.-H.P. and M.C. designed the MPAO-based two-photon imaging system. J.-H.P. developed the wavefront measurement and correction algorithm, implemented the imaging system, and performed structural imaging of neurons and dynamic imaging of microglia. L.K. designed and performed structural imaging and calcium imaging of neurons, dynamic imaging of microglia, and nonplanar imaging of blood vessels and neurons (data shown in Figs. 1,2,3 and Supplementary Figs. 6,9,10,11,12). Y.Z. supported the system development, performed structural imaging of neurons and microglia, and assisted with figure preparation. M.C. and L.K. wrote the manuscript with input from all authors.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Meng Cui.

Integrated supplementary information

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Figures 1–14, Supplementary Notes 1–4 and Supplementary Table 1.

Zip files

  1. 1.

    Supplementary Software

    Matlab code for wavefront measurement.

Videos

  1. 1.

    Volumetric imaging of microglia with different wavefront correction methods

    We show the MIP of microglia along z axis (450 × 450 × 40 μm3 volume at the depth of 330-370 μm) recorded by system correction with MPAO, full correction with MPAO, full correction with an averaged wavefront (full average), and full correction with the wavefront measured from one region (full single, nine groups in sequence).

  2. 2.

    Dynamic imaging of resting state microglia, dendritic cells and other leukocytes

    MIP along z axis of the 450 × 450 × 50 μm3 volume at 0-50 μm under the dura. Using MPAO with full correction, we performed large FOV volumetric imaging of resting state microglia. In addition to the motion of microglia and dendritic cells, the video also captured GFP-expressing leukocytes trafficking in brain vasculatures.

  3. 3.

    Dynamic imaging of microglia before and after the activation by laser ablation

    MIP along z axis of the 450 × 450 × 35 μm3 volume at the depth of 330-365 μm. Using laser ablation, we introduced tissue damage, and employed MPAO with full correction to record the activated microglia dynamics.

  4. 4.

    3D rendering of MPAO recorded layer 5 neurons

    The image volume was 450×450×550 μm3. We show the volume data with system and with full correction for comparison.

  5. 5.

    Large-FOV calcium imaging at 15 Hz with full correction by MPAO

    In the video, we show the partial data of the calcium transient in Supplementary Fig. 10. The imaging FOV is 450 × 450 μm 2.

  6. 6.

    3D image stack with the depth information of each image segment in the nonplanar image recording

    The image in Supplementary Fig. 11d is extracted from this video. Magenta: the 3D image stacks. Green: the planes defined by the defocusing wavefront in nonplanar imaging. The imaging FOV is 450 × 450 μm2.

  7. 7.

    Nonplanar imaging of 3D neurovasculature regulation

    The plot in Fig. 3e is extracted from this video. The imaging FOV is 450 × 450 μm 2.

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DOI

https://doi.org/10.1038/nmeth.4290

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