Wide-field three-photon excitation in biological samples

Christopher J Rowlands; Demian Park; Oliver T Bruns; Kiryl D Piatkevich; Dai Fukumura; Rakesh K Jain; Moungi G Bawendi; Edward S Boyden; Peter TC So

doi:10.1038/lsa.2016.255

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Wide-field three-photon excitation in biological samples

Original Articles

- Wide-field three-photon excitation in biological samples
- Wide-field three-photon excitation in biological samples
- LSA 2017年6卷第2期
- Affiliations：
  
  1.Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA
  2.Media Lab, Massachusetts Institute of Technology, Cambridge, MA 02139, USA
  3.Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA 02139, USA
  4.Edwin L. Steele Laboratories, Department of Radiation Oncology, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA
  5.Department of Brain and Cognitive Sciences, McGovern Institute and MIT Center for Neurobiological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139-4307, USA
  6.Department of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA
- Author bio：
  
  C Rowlands, Email: chrisrow@mit.edu
- Funds：
- DOI：10.1038/lsa.2016.255
  中图分类号：
- 纸质出版日期：2017-06，
  
  网络出版日期：2016-11-23，
  
  收稿日期：2016-01-22，
  
  修回日期：2016-11-17，
  
  录用日期：2016-11-21
- Accepted：
Scan QR Code
Wide-field three-photon excitation in biological samples[J]. LSA, 2017,6(2).

Rowlands, C. J. et al. Wide-field three-photon excitation in biological samples. Light: Science & Applications, 6, e16255 (2017).
Wide-field three-photon excitation in biological samples[J]. LSA, 2017,6(2). DOI： 10.1038/lsa.2016.255.

Rowlands, C. J. et al. Wide-field three-photon excitation in biological samples. Light: Science & Applications, 6, e16255 (2017). DOI： 10.1038/lsa.2016.255.

摘要

Abstract

Three-photon wide-field depth-resolved excitation is used to overcome some of the limitations in conventional point-scanning two- and three-photon microscopy. Excitation of chromophores as diverse as channelrhodopsins and quantum dots is shown

and a penetration depth of more than 700 μm into fixed scattering brain tissue is achieved

approximately twice as deep as that achieved using two-photon wide-field excitation. Compatibility with live animal experiments is confirmed by imaging the cerebral vasculature of an anesthetized mouse; a complete focal stack was obtained without any evidence of photodamage. As an additional validation of the utility of wide-field three-photon excitation

functional excitation is demonstrated by performing three-photon optogenetic stimulation of cultured mouse hippocampal neurons expressing a channelrhodopsin; action potentials could reliably be excited without causing photodamage.

关键词

Keywords

biophotonicsmultiphoton microscopyoptogeneticstemporal focusingthree-photon

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