Two-color multiphoton <italic style="font-style: italic">in vivo</italic> imaging with a femtosecond diamond Raman laser

Evan P Perillo; Jeremy W Jarrett; Yen-Liang Liu; Ahmed Hassan; Daniel C Fernée; John R Goldak; Andrei Bonteanu; David J Spence; Hsin-Chih Yeh; Andrew K Dunn

doi:10.1038/lsa.2017.95

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Two-color multiphoton in vivo imaging with a femtosecond diamond Raman laser

Original Articles

- Two-color multiphoton in vivo imaging with a femtosecond diamond Raman laser
- Light: Science & Applications Vol. 6, Issue 4, (2017)
- Affiliations：
  
  1.Department of Biomedical Engineering, The University of Texas at Austin, TX 78712, USA
  2.Department of Physics, The University of Texas at Austin, TX 78712, USA
  3.MQ Photonics, Department of Physics and Astronomy, Macquarie University, Sydney, NSW 2109, Australia
- Author bio：
  
  AK Dunn, Email: adunn@utexas.edu
- Funds：
- DOI：10.1038/lsa.2017.95
  CLC：
- Published：2017-12，
  
  Published Online：01 June 2017，
  
  Received：21 December 2016，
  
  Revised：22 May 2017，
  
  Accepted：31 May 2017
- Accepted：
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Perillo, E. P. et al. Two-color multiphoton in vivo imaging with a femtosecond diamond Raman laser. Light: Science & Applications, 6, e17095 (2017).
DOI：

Perillo, E. P. et al. Two-color multiphoton in vivo imaging with a femtosecond diamond Raman laser. Light: Science & Applications, 6, e17095 (2017). DOI： 10.1038/lsa.2017.95.

摘要

Abstract

Two-color multiphoton microscopy through wavelength mixing of synchronized lasers has been shown to increase the spectral window of excitable fluorophores without the need for wavelength tuning. However

most currently available dual output laser sources rely on the costly and complicated optical parametric generation approach. In this report

we detail a relatively simple and low cost diamond Raman laser pumped by a ytterbium fiber amplifier emitting at 1055 nm

which generates a first Stokes emission centered at 1240 nm with a pulse width of 100 fs. The two excitation wavelengths of 1055 and 1240 nm

along with the effective two-color excitation wavelength of 1140 nm

provide an almost complete coverage of fluorophores excitable within the range of 1000–1300 nm. When compared with 1055 nm excitation

two-color excitation at 1140 nm offers a 90% increase in signal for many far-red emitting fluorescent proteins (for ex

ample

tdKatushka2). We demonstrate multicolor imaging of tdKatushka2 and Hoechst 33342 via simultaneous two-color two-photon

and two-color three-photon microscopy in engineered 3D multicellular spheroids. We further discuss potential benefits and applications for two-color three-photon excitation. In addition

we show that this laser system is capable of

in vivo

imaging in mouse cortex to nearly 1 mm in depth with two-color excitation.

关键词

Keywords

microscopynonlinear processesthree-dimensional imagingultrafast lasers

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⁰

Two-color multiphoton in vivo imaging with a femtosecond diamond Raman laser

DOI：10.1038/lsa.2017.95

摘要

Abstract

关键词

Keywords

references