Cited by 1,505Advanced Fluorescence Microscopy TechniquesOptical Imaging and Spectroscopy TechniquesNonlinear Optical Materials Studies

In vivo three-photon microscopy of subcortical structures within an intact mouse brain

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Horton, Nicholas G., Wang, Ke, Kobat, Demirhan, Clark, Catharine G., Wise, Frank W., Schaffer, Chris B., Xu, Chris·2013·Nature Photonics

Abstract

Two-photon fluorescence microscopy (2PM)1 enables scientists in various fields including neuroscience 2,3, embryology 4, and oncology 5 to visualize in vivo and ex vivo tissue morphology and physiology at a cellular level deep within scattering tissue. However, tissue scattering limits the maximum imaging depth of 2PM within the mouse brain to the cortical layer, and imaging subcortical structures currently requires the removal of overlying brain tissue 3 or the insertion of optical probes 6,7. Here we demonstrate non-invasive, high resolution, in vivo imaging of subcortical structures within an intact mouse brain using three-photon fluorescence microscopy (3PM) at a spectral excitation window of 1,700 nm. Vascular structures as well as red fluorescent protein (RFP)-labeled neurons within the mouse hippocampus are imaged. The combination of the long excitation wavelength and the higher order nonlinear excitation overcomes the limitations of 2PM, enabling biological investigations to take place at greater depth within tissue.

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Crossref · 10.1038/nphoton.2012.336

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  1. [27]Franklin KBJ Paxinos G Mouse brain in stereotaxic coordinates Academic Press London 2008

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Authors
Horton, N. G., Wang, K., Kobat, D., Clark, C. G., Wise, F. W., Schaffer, C. B., & Xu, C.
Affiliations
Cornell University (US)
Corresponding author
Xu, Chris
Venue
Nature Photonics
Published
2013-01-20
Volume
7
Issue
3
Pages
205-209
ISSN
  • 1749-4885 (ISSN-L)
  • 1749-4893
OpenAlex
W2150708165
Cited by
1,505
References
32
Citations by year
2026: 84 · 2025: 102 · 2024: 136 · 2023: 98 · 2022: 146 · 2021: 126 · 2020: 123 · 2019: 122 · 2018: 119 · 2017: 127 · 2016: 105 · 2015: 111 · 2014: 70 · 2013: 36
FWCI
65.07
Type
journal article
Language
en
Open access
Locations
  • Nature Photonics (publishedVersion)
  • PubMed (publishedVersion)
  • Europe PMC (PubMed Central) (acceptedVersion)
  • PubMed Central (acceptedVersion)
Funders
  • National Science Foundation
  • National Institutes of Health
  • National Cancer Institute (R01CA133148)
  • National Institute of General Medical Sciences (R21GM103461)
  • National Institute of Biomedical Imaging and Bioengineering (R01EB014873)
  • National Center for Research Resources (R21RR032392)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
  • Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Optical Imaging and Spectroscopy Techniques
  • Physical Sciences › Engineering › Biomedical Engineering › Nonlinear Optical Materials Studies
Keywords
three-photon microscopy, mouse brain, hippocampus, deep tissue imaging, tissue scattering, vascular imaging, infrared excitation, noninvasive in vivo imaging

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