Miniscope3D: optimized single-shot miniature 3D fluorescence microscopy

Kyrollos Yanny; Nick Antipa; William Liberti; Sam Dehaeck; Kristina Monakhova; Fanglin Linda Liu; Konlin Shen; Ren Ng; Laura Waller

doi:10.1038/s41377-020-00403-7

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Miniscope3D: optimized single-shot miniature 3D fluorescence microscopy

Original Articles

- Miniscope3D: optimized single-shot miniature 3D fluorescence microscopy
- Light: Science & Applications Vol. 9, Issue 5, Pages: 1638-1650(2020)
- Affiliations：
  
  1.UCB/UCSF Joint Graduate Program in Bioengineering, University of California, Berkeley, CA 94720, USA
  2.Department of Electrical Engineering & Computer Sciences, University of California, Berkeley, CA 94720, USA
  3.TIPs Department, Université libre de Bruxelles (ULB), 1050 Brussels, Belgium
- Author bio：
  
  Kyrollos Yanny (kyrollosyanny@gmail.com)
  Nick Antipa (naantipa@gmail.com)
- Funds：
- DOI：10.1038/s41377-020-00403-7
  CLC：
- Published：2020，
  
  Published Online：02 October 2020，
  
  Received：12 May 2020，
  
  Revised：01 September 2020，
  
  Accepted：04 September 2020
- Accepted：
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Yanny, K. et al. Miniscope3D: optimized single-shot miniature 3D fluorescence microscopy. Light: Science & Applications, 9, 1638-1650 (2020).
DOI：

Yanny, K. et al. Miniscope3D: optimized single-shot miniature 3D fluorescence microscopy. Light: Science & Applications, 9, 1638-1650 (2020). DOI： 10.1038/s41377-020-00403-7.

摘要

Abstract

Miniature fluorescence microscopes are a standard tool in systems biology. However

widefield miniature microscopes capture only 2D information

and modifications that enable 3D capabilities increase the size and weight and have poor resolution outside a narrow depth range. Here

we achieve the 3D capability by replacing the tube lens of a conventional 2D Miniscope with an optimized multifocal phase mask at the objective's aperture stop. Placing the phase mask at the aperture stop significantly reduces the size of the device

and varying the focal lengths enables a uniform resolution across a wide depth range. The phase mask encodes the 3D fluorescence intensity into a single 2D measurement

and the 3D volume is recovered by solving a sparsity-constrained inverse problem. We provide methods for designing and fabricating the phase mask and an efficient forward model that accounts for the field-varying aberrations in miniature objectives. We demonstrate a prototype that is 17 mm tall and weighs 2.5 grams

achieving 2.76 μm lateral

and 15 μm axial resolution across most of the 900 × 700 × 390 μm

volume at 40 volumes per second. The performance is validated experimentally on resolution targets

dynamic biological samples

and mouse brain tissue. Compared with existing miniature single-shot volume-capture implementations

our system is smaller and lighter and achieves a more than 2× better lateral and axial resolution throughout a 10× larger usable depth range. Our microscope design provides single-shot 3D imaging for applications where a compact platform matters

such as volumetric neural imaging in freely moving animals and 3D motion studies of dynamic samples in incubators and lab-on-a-chip devices.

关键词

Keywords

references

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