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Speed scaling in multiphoton fluorescence microscopy

Nature Photonics volume 15, pages 800–812 (2021)Cite this article

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A Publisher Correction to this article was published on 29 November 2021

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Abstract

After over 30 years of advances, multiphoton microscopy (MPM) is now instrumental in a wide range of in vivo biological imaging applications. However, it has, until recently, remained not achievable or affordable to meet the unmet need for fast monitoring of biological dynamics and large-scale examination of biological heterogeneity. Only within the past few years have new strategies emerged to empower MPM at a speed that was once inconceivable, notably at kilohertz two-dimensional (2D) frame rate, and 3D rate or beyond. This Review highlights the latest high-speed innovations and discusses the potential of their synergism with other advanced, but less speed-centric MPM toolboxes. Recognizing these prospects and challenges could inspire new approaches for reprioritizing imaging speed in future MPM developments.

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Fig. 1: Advances in high-speed 2PFM.
Fig. 2: High-speed strategies enabling planar MPM at subkilohertz to kilohertz frame rate.
Fig. 3: Strategies for high-speed MPM axial access.
Fig. 4: Ongoing advances in MPM that address the challenges of imaging a FOV, SNR and tissue penetration, together with temporal and spatial resolution.

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Acknowledgements

This work is supported by the Research Grants Council of the Hong Kong Special Administrative Region of China (grants 17208918, 17209017 and 17259316 to J.W. and K.K.T.; RFS2021-7S06 and C7047-16G to K.K.T.) and NIH BRAIN Initiative grants (1UF1NS107696 to J.W., N.J. and K.K.T.).

Author information

Authors and Affiliations

  1. Department of Physics, University of California, Berkeley, CA, USA

    Jianglai Wu & Na Ji

  2. Chinese Institute for Brain Research, Beijing, China

    Jianglai Wu

  3. Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA

    Na Ji

  4. Department of Molecular and Cell Biology, University of California, Berkeley, CA, USA

    Na Ji

  5. Helen Wills Neuroscience Institute, University of California, Berkeley, CA, USA

    Na Ji

  6. Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, CA, USA

    Na Ji

  7. Department of Electrical and Electronic Engineering, The University of Hong Kong, Hong Kong, China

    Kevin K. Tsia

  8. Advanced Biomedical Instrumentation Centre, Hong Kong Science Park, Shatin, Hong Kong, China

    Kevin K. Tsia

Authors
  1. Jianglai Wu
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  2. Na Ji
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  3. Kevin K. Tsia
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Contributions

All authors contributed to writing the manuscript.

Corresponding authors

Correspondence to Na Ji or Kevin K. Tsia.

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The authors declare no competing interests.

Additional information

Peer review information Nature Photonics thanks Keisuke Goda and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Wu, J., Ji, N. & Tsia, K.K. Speed scaling in multiphoton fluorescence microscopy. Nat. Photon. 15, 800–812 (2021). https://doi.org/10.1038/s41566-021-00881-0

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