Integrated Fluorescent Nanoprobe Design for High‐Speed In Vivo Two‐Photon Microscopic Imaging of Deep‐Brain Vasculature in Mice

2021 ◽  
pp. 2010698
Author(s):  
Minami Takezaki ◽  
Ryosuke Kawakami ◽  
Shozo Onishi ◽  
Yasutaka Suzuki ◽  
Jun Kawamata ◽  
...  
Keyword(s):  
High Speed ◽  
Microscopic Imaging ◽  
Two Photon ◽  
Fluorescent Nanoprobe ◽  
Brain Vasculature ◽  
Deep Brain

scholarly journals AIE-active two-photon fluorescent nanoprobe with NIR-II light excitability for highly efficient deep brain vasculature imaging

Theranostics ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 2137-2148
Author(s):  
Soham Samanta ◽  
Meina Huang ◽  
Shaoqiang Li ◽  
Zhigang Yang ◽  
Ying He ◽  
...  
Keyword(s):  
Two Photon ◽  
Fluorescent Nanoprobe ◽  
Highly Efficient ◽  
Brain Vasculature ◽  
Deep Brain

scholarly journals High-speed volumetric two-photon fluorescence imaging of neurovascular dynamics

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Jiang Lan Fan ◽  
Jose A. Rivera ◽  
Wei Sun ◽  
John Peterson ◽  
Henry Haeberle ◽  
...  
Keyword(s):  
Blood Flow ◽  
High Speed ◽  
Laser Scanning ◽  
Three Dimensional ◽  
Laser Scanning Microscopy ◽  
Fluorescence Signal ◽  
Imaging Method ◽  
Spatiotemporal Resolution ◽  
Two Photon

AbstractUnderstanding the structure and function of vasculature in the brain requires us to monitor distributed hemodynamics at high spatial and temporal resolution in three-dimensional (3D) volumes in vivo. Currently, a volumetric vasculature imaging method with sub-capillary spatial resolution and blood flow-resolving speed is lacking. Here, using two-photon laser scanning microscopy (TPLSM) with an axially extended Bessel focus, we capture volumetric hemodynamics in the awake mouse brain at a spatiotemporal resolution sufficient for measuring capillary size and blood flow. With Bessel TPLSM, the fluorescence signal of a vessel becomes proportional to its size, which enables convenient intensity-based analysis of vessel dilation and constriction dynamics in large volumes. We observe entrainment of vasodilation and vasoconstriction with pupil diameter and measure 3D blood flow at 99 volumes/second. Demonstrating high-throughput monitoring of hemodynamics in the awake brain, we expect Bessel TPLSM to make broad impacts on neurovasculature research.

scholarly journals Dual-color volumetric imaging of neural activity of cortical columns

2018 ◽  
Author(s):  
Shuting Han ◽  
Weijian Yang ◽  
Rafael Yuste
Keyword(s):  
Neural Activity ◽  
High Speed ◽  
Neural Circuits ◽  
Emergent Properties ◽  
Spatiotemporal Resolution ◽  
Population Activity ◽  
Volumetric Imaging ◽  
Two Photon ◽  
Dual Color

To capture the emergent properties of neural circuits, high-speed volumetric imaging of neural activity at cellular resolution is desirable. But while conventional two-photon calcium imaging is a powerful tool to study population activity in vivo, it is restrained to two-dimensional planes. Expanding it to 3D while maintaining high spatiotemporal resolution appears necessary. Here, we developed a two-photon microscope with dual-color laser excitation that can image neural activity in a 3D volume. We imaged the neuronal activity of primary visual cortex from awake mice, spanning from L2 to L5 with 10 planes, at a rate of 10 vol/sec, and demonstrated volumetric imaging of L1 long-range PFC projections and L2/3 somatas. Using this method, we map visually-evoked neuronal ensembles in 3D, finding a lack of columnar structure in orientation responses and revealing functional correlations between cortical layers which differ from trial to trial and are missed in sequential imaging. We also reveal functional interactions between presynaptic L1 axons and postsynaptic L2/3 neurons. Volumetric two-photon imaging appears an ideal method for functional connectomics of neural circuits.

44 nJ, 114 fs Nd-doped fiber laser pulses at 920nm for in vivo two-photon microscopic imaging

CLEO: 2015 ◽  
2015 ◽  
Author(s):  
Bingying Chen ◽  
Tongxiao Jiang ◽  
Weijian Zong ◽  
Fuzeng Niu ◽  
Liangyi Chen ◽  
...  
Keyword(s):  
Fiber Laser ◽  
Laser Pulses ◽  
Microscopic Imaging ◽  
Two Photon

scholarly journals 910nm femtosecond Nd-doped fiber laser for in vivo two-photon microscopic imaging

2016 ◽  
Vol 24 (15) ◽  
pp. 16544 ◽  
Author(s):  
Bingying Chen ◽  
Tongxiao Jiang ◽  
Weijian Zong ◽  
Liangyi Chen ◽  
Zhigang Zhang ◽  
...  
Keyword(s):  
Fiber Laser ◽  
Microscopic Imaging ◽  
Two Photon

scholarly journals Adaptive optics two-photon endomicroscopy enables deep-brain imaging at synaptic resolution over large volumes

2020 ◽  
Vol 6 (40) ◽  
pp. eabc6521 ◽  
Author(s):  
Zhongya Qin ◽  
Congping Chen ◽  
Sicong He ◽  
Ye Wang ◽  
Kam Fai Tam ◽  
...  
Keyword(s):  
Brain Imaging ◽  
Adaptive Optics ◽  
Critical Role ◽  
Random Access ◽  
Brain Structures ◽  
Invasive Approach ◽  
Two Photon ◽  
Grin Lens ◽  
Deep Brain

Optical deep-brain imaging in vivo at high resolution has remained a great challenge over the decades. Two-photon endomicroscopy provides a minimally invasive approach to image buried brain structures, once it is integrated with a gradient refractive index (GRIN) lens embedded in the brain. However, its imaging resolution and field of view are compromised by the intrinsic aberrations of the GRIN lens. Here, we develop a two-photon endomicroscopy by adding adaptive optics based on direct wavefront sensing, which enables recovery of diffraction-limited resolution in deep-brain imaging. A new precompensation strategy plays a critical role to correct aberrations over large volumes and achieve rapid random-access multiplane imaging. We investigate the neuronal plasticity in the hippocampus, a critical deep brain structure, and reveal the relationship between the somatic and dendritic activity of pyramidal neurons.

scholarly journals Photon counting, censor corrections, and lifetime imaging for improved detection in two-photon microscopy

2011 ◽  
Vol 105 (6) ◽  
pp. 3106-3113 ◽  
Author(s):  
Jonathan D. Driscoll ◽  
Andy Y. Shih ◽  
Satish Iyengar ◽  
Jeffrey J. Field ◽  
G. Allen White ◽  
...  
Keyword(s):  
High Speed ◽  
Signal To Noise Ratio ◽  
Photon Counting ◽  
In Vivo Model ◽  
Fluorescence Lifetime Imaging ◽  
Photon Counter ◽  
Two Photon Microscopy ◽  
Two Photon ◽  
Lifetime Imaging

We present a high-speed photon counter for use with two-photon microscopy. Counting pulses of photocurrent, as opposed to analog integration, maximizes the signal-to-noise ratio so long as the uncertainty in the count does not exceed the gain-noise of the photodetector. Our system extends this improvement through an estimate of the count that corrects for the censored period after detection of an emission event. The same system can be rapidly reconfigured in software for fluorescence lifetime imaging, which we illustrate by distinguishing between two spectrally similar fluorophores in an in vivo model of microstroke.

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