Converting lateral scanning into axial focusing to speed up three-dimensional microscopy

Tonmoy Chakraborty; Bingying Chen; Stephan Daetwyler; Bo-Jui Chang; Oliver Vanderpoorten; Etai Sapoznik; Clemens F. Kaminski; Tuomas P. J. Knowles; Kevin M. Dean; Reto Fiolka

doi:10.1038/s41377-020-00401-9

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Converting lateral scanning into axial focusing to speed up three-dimensional microscopy

Original Articles

- Converting lateral scanning into axial focusing to speed up three-dimensional microscopy
- Light: Science & Applications Vol. 9, Issue 5, Pages: 1587-1598(2020)
- Affiliations：
  
  1.Department of Cell Biology, UT Southwestern Medical Center, Dallas, TX, USA
  2.Department of Physics and Astronomy, University of New Mexico, Albuquerque, NM, USA
  3.Department of Chemical Engineering and Biotechnology, University of Cambridge, Cambridge CB3 0AS, UK
  4.Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, UK
  5.Lyda Hill Department of Bioinformatics, UT Southwestern Medical Center, Dallas, TX, USA
- Author bio：
  
  Reto Fiolka (Reto.Fiolka@utsouthwestern.edu)
- Funds：
- DOI：10.1038/s41377-020-00401-9
  CLC：
- Published：2020，
  
  Published Online：18 September 2020，
  
  Received：18 March 2020，
  
  Revised：25 August 2020，
  
  Accepted：02 September 2020
- Accepted：
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Chakraborty, T. et al. Converting lateral scanning into axial focusing to speed up three-dimensional microscopy. Light: Science & Applications, 9, 1587-1598 (2020).
DOI：

Chakraborty, T. et al. Converting lateral scanning into axial focusing to speed up three-dimensional microscopy. Light: Science & Applications, 9, 1587-1598 (2020). DOI： 10.1038/s41377-020-00401-9.

摘要

Abstract

In optical microscopy

the slow axial scanning rate of the objective or the sample has traditionally limited the speed of volumetric imaging. Recently

by conjugating either a movable mirror to the image plane in a remote-focusing geometry or an electrically tuneable lens (ETL) to the back focal plane

rapid axial scanning has been achieved. However

mechanical actuation of a mirror limits the axial scanning rate (usually only 10–100 Hz for piezoelectric or voice coil-based actuators)

while ETLs introduce spherical and higher-order aberrations that prevent high-resolution imaging. In an effort to overcome these limitations

we introduce a novel optical design that transforms a lateral-scan motion into a spherical aberration-free axial scan that can be used for high-resolution imaging. Using a galvanometric mirror

we scan a laser beam laterally in a remote-focusing arm

which is then back-reflected from different heights of a mirror in the image space. We characterize the optical performance of this remote-focusing technique and use it to accelerate axially swept light-sheet microscopy by an order of magnitude

allowing the quantification of rapid vesicular dynamics in three dimensions. We also demonstrate resonant remote focusing at 12 kHz with a two-photon raster-scanning microscope

which allows rapid imaging of brain tissues and zebrafish cardiac dynamics with diffraction-limited resolution.

关键词

Keywords

references

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