Image scanning microscopy based on multifocal metalens for sub-diffraction-limited imaging of brain organoids

Yongjae Jo; Hyemi Park; Seho Lee; Hyeyoung Yoon; Taehoon Lee; Gyusoo Bak; Hanjun Cho; Jong-Chan Park; Inki Kim

doi:10.1038/s41377-025-01900-3

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Image scanning microscopy based on multifocal metalens for sub-diffraction-limited imaging of brain organoids

Original Articles

- Image scanning microscopy based on multifocal metalens for sub-diffraction-limited imaging of brain organoids
- Light: Science & Applications Vol. 14, Issue 11, Pages: 3765-3780(2025)
- Affiliations：
  
  1.Department of Biophysics, Institute of Quantum Biophysics, Sungkyunkwan University, Suwon, Republic of Korea
  2.Department of Intelligent Precision Healthcare Convergence, Sungkyunkwan University, Suwon, Republic of Korea
  3.Center for Quantum Technology, Korea Institute of Science and Technology (KIST), Seoul, Republic of Korea
  4.Department of Biopharmaceutical Convergence, Sungkyunkwan University, Suwon, Republic of Korea
  5.Department of MetaBioHealth, Sungkyunkwan University, Suwon, Republic of Korea
- Author bio：
  
  Inki Kim (inki.kim@skku.edu)
- Funds：
- DOI：10.1038/s41377-025-01900-3
  CLC：
- Received：19 November 2024，
  
  Revised：2025-05-16，
  
  Accepted：21 May 2025，
  
  Published Online：13 October 2025，
  
  Published：30 November 2025
- Accepted：
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Jo, Y. et al. Image scanning microscopy based on multifocal metalens for sub-diffraction-limited imaging of brain organoids. Light: Science & Applications, 14, 3765-3780 (2025).
DOI：

Jo, Y. et al. Image scanning microscopy based on multifocal metalens for sub-diffraction-limited imaging of brain organoids. Light: Science & Applications, 14, 3765-3780 (2025). DOI： 10.1038/s41377-025-01900-3.

摘要

Abstract

Image scanning microscopy (ISM) is a promising imaging technique that offers sub-diffraction-limited

resolution and optical sectioning. Theoretically

ISM can improve the optical resolution by a factor of two through pixel reassignment and deconvolution. Multifocal array illumination and scanning have been widely adopted to implement ISM because of their simplicity. Conventionally

digital micromirror devices (DMDs)

and microlens arrays (MLAs)

have been used to generate dense and uniform multifocal arrays for ISM

which are critical for achieving fast imaging and high-quality ISM reconstruction. However

these approaches have limitations in terms of cost

numerical aperture (NA)

pitch

and uniformity

making it challenging to create dense and high-quality multifocal arrays at high NA. To overcome these limitations

we introduced a novel multifocal metalens design strategy called the hybrid multiplexing method

which combines two conventional multiplexing approaches: phase addition and random multiplexing. Through numerical simulations

we demonstrate that the proposed method generates more uniform and denser multifocal arrays than conventional methods

even at small pitches. As a proof of concept

we fabricated a multifocal metalens generating 40 × 40 array of foci with a 3 μm pitch and NA of 0.7 operating at a wavelength of 488 nm and then constructed the multifocal metalens-based ISM (MMISM). We demonstrated that MMISM successfully resolved sub-diffraction-limited features in imaging of microbead samples and forebrain organoid sections. The results showed that MMISM imaging achieved twice the diffraction-limited resolution and revealed clearer structural features of neurons compared to wide-field images. We anticipate that our novel design strategy can be widely applied to produce multifunctional optical elements and replace conventional optical elements in specialized applications.

关键词

Keywords

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