Open Access
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
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TLDR
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.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.read more
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Deep optical imaging within complex scattering media
Seokchan Yoon,Moonseok Kim,Mooseok Jang,Mooseok Jang,Youngwoon Choi,Wonjun Choi,Sungsam Kang,Wonshik Choi +7 more
TL;DR: In this paper, the authors provide an overview of recent developments in this area, with a focus on approaches that achieve a microscopic spatial resolution while remaining useful for in vivo imaging, and discuss their present limitations and future prospects.
Journal ArticleDOI
Light-sheet microscopy in the near-infrared II window.
Feifei Wang,Hao Wan,Zhuoran Ma,Yeteng Zhong,Qinchao Sun,Ye Tian,Liangqiong Qu,Haotian Du,Mingxi Zhang,Lulin Li,Huilong Ma,Jian Luo,Jian Luo,Yongye Liang,Wen J. Li,Wen J. Li,Guosong Hong,Lianqing Liu,Hongjie Dai +18 more
TL;DR: Near-infrared II light-sheet microscopy in normal and oblique configurations enabled in vivo imaging of live mice through intact tissue, revealing abnormal blood flow and T-cell motion in tumor microcirculation and mapping out programmed-death ligand 1 and programmed cell death protein 1 in tumors with cellular resolution.
Journal ArticleDOI
A review of biomedical multiphoton microscopy and its laser sources
TL;DR: In this article, the main interests and technical requirements of MPM are resumed with a focus on the crucial role of excitation source for optimization of multiphoton processes, and an overview of the different sources successfully demonstrated in the literature for MPM is presented and their physical parameters are inventoried.
Journal ArticleDOI
Three-photon neuronal imaging in deep mouse brain
Tianyu Wang,Chris Xu +1 more
TL;DR: A systematic summary of the optimization of three-photon imaging parameters for neural imaging, based on their effects on calcium imaging quality and perturbation to brain tissues, is presented.
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Dual-color deep-tissue three-photon microscopy with a multiband infrared laser
Khmaies Guesmi,Lamiae Abdeladim,Samuel Tozer,Pierre Mahou,Takuma Kumamoto,Karolis Jurkus,Philippe Rigaud,Karine Loulier,Nicolas Dray,Patrick Georges,Marc Hanna,Jean Livet,Willy Supatto,Emmanuel Beaurepaire,Frédéric Druon +14 more
TL;DR: In-depth dual-color 3P imaging in a fixed mouse brain, chick embryo spinal cord, and live adult zebrafish brain is demonstrated, with an improved signal-to-background ratio compared to multicolor two-photon imaging.
References
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TL;DR: 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.