In vivo three-photon microscopy of subcortical structures within an intact mouse brain
Nicholas G. Horton,Ke Wang,Demirhan Kobat,Catharine G. Clark,Frank W. Wise,Chris B. Schaffer,Chris Xu +6 more
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TLDR
Non-invasive, high-resolution, in vivo imaging of subcortical structures (the external capsule and hippocampus) within an intact mouse brain is demonstrated using three-photon fluorescence microscopy at the new spectral window of 1700 nm.Abstract:
Two-photon fluorescence microscopy (2PM)1 enables scientists in various fields including neuroscience2,3, embryology4, and oncology5 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 tissue3 or the insertion of optical probes6,7. 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.read more
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Improved deep two-photon calcium imaging in vivo.
TL;DR: The method involves an optimized protocol for depth-restricted labeling with the red-shifted fluorescent calcium indicator Cal-590 and benefits from the use of ultra-short laser pulses, which allows in vivo functional imaging of neuronal populations with single cell resolution in all six layers of the mouse cortex.
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Octave-spanning supercontinuum generation in a silicon-rich nitride waveguide
Xing Liu,Minhao Pu,Binbin Zhou,Clemens J. Kruckel,Attila Fülöp,Victor Torres-Company,Morten Bache +6 more
TL;DR: This work generates supercontinuum (817-2250 nm at -30dB) in a dispersion-engineered silicon-rich nitride waveguide by pumping fs pulses with 82 pJ from an erbium-fiber oscillator.
Journal ArticleDOI
Investigation of the long wavelength limit of soliton self-frequency shift in a silica fiber.
TL;DR: In vivo two-photon excited fluorescence imaging of vasculature and neurons in a mouse brain at wavelength beyond the tuning range of a mode-locked Ti:Sapphire lasers is demonstrated.
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 T. C. So +8 more
TL;DR: In this article, three-photon wide-field depth-resolved excitation was used to overcome some of the limitations in conventional point-scanning 2-and 3-Photon microscopy, achieving a penetration depth of more than 700 μm into fixed scattering brain tissue.
Journal ArticleDOI
High-resolution structural and functional deep brain imaging using adaptive optics three-photon microscopy
Lina L. Streich,Lina L. Streich,Juan Carlos Boffi,Ling Wang,Khaleel Alhalaseh,Matteo Barbieri,Ronja Rehm,Senthilkumar Deivasigamani,Cornelius Gross,Amit Agarwal,Robert Prevedel +10 more
TL;DR: In this paper, a minimally invasive intravital imaging methodology based on three-photon excitation, indirect adaptive optics (AO) and active electrocardiogram (ECG) gating was proposed to advance deep-tissue imaging.
References
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Book
The Mouse Brain in Stereotaxic Coordinates
TL;DR: The 3rd edition of this atlas is now in more practical 14"x11" format for convenient lab use and includes a CD of all plates and diagrams, as well as Adobe Illustrator files of the diagrams, and a variety of additional useful material.
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Journal ArticleDOI
Deep tissue two-photon microscopy
Fritjof Helmchen,Winfried Denk +1 more
TL;DR: Fundamental concepts of nonlinear microscopy are reviewed and conditions relevant for achieving large imaging depths in intact tissue are discussed.
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Measurement of two-photon excitation cross sections of molecular fluorophores with data from 690 to 1050 nm
Chris Xu,Watt W. Webb +1 more
TL;DR: In this paper, the two-photon fluorescence excitation (TPE) spectra were measured for 11 common molecular fluorophores in the excitation wavelength range 690 nm < λ < 1050 nm.
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Transgenic strategies for combinatorial expression of fluorescent proteins in the nervous system
Jean Livet,Tamily A. Weissman,Hyuno Kang,Ju Lu,Robyn A Bennis,Joshua R. Sanes,Jeff W. Lichtman +6 more
TL;DR: Strategies to visualize synaptic circuits by genetically labelling neurons with multiple, distinct colours are presented and may facilitate the analysis of neuronal circuitry on a large scale.