scholarly journals Volumetric Two-photon Imaging of Neurons Using Stereoscopy (vTwINS)

2016 ◽  
Author(s):  
Alexander Song ◽  
Adam S. Charles ◽  
Sue Ann Koay ◽  
Jeff L. Gauthier ◽  
Stephan Y. Thiberge ◽  
...  
Keyword(s):  
Calcium Imaging ◽  
Laser Scanning ◽  
Large Scale ◽  
Laser Scanning Microscopy ◽  
Frame Rate ◽  
Neural Population ◽  
Imaging Method ◽  
Two Photon ◽  
Photon Imaging ◽  
Two Photon Imaging

AbstractTwo-photon laser scanning microscopy of calcium dynamics using fluorescent indicators is a widely used imaging method for large scale recording of neural activity in vivo. Here we introduce volumetric Two-photon Imaging of Neurons using Stereoscopy (vTwINS), a volumetric calcium imaging method that employs an elongated, V-shaped point spread function to image a 3D brain volume. Single neurons project to spatially displaced image pairs in the resulting 2D image, and the separation distance between images is proportional to depth in the volume. To demix the fluorescence time series of individual neurons, we introduce a novel orthogonal matching pursuit algorithm that also infers source locations within the 3D volume. We illustrate vTwINS by imaging neural population activity in mouse primary visual cortex and hippocampus. Our results demonstrate that vTwINS provides an effective method for volumetric two-photon calcium imaging that increases the number of neurons recorded while maintaining a high frame-rate.

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 Complex Pattern Selectivity in Macaque Primary Visual Cortex Revealed by Large-Scale Two-Photon Imaging

2018 ◽  
Vol 28 (1) ◽  
pp. 38-48.e3 ◽  
Author(s):  
Shiming Tang ◽  
Tai Sing Lee ◽  
Ming Li ◽  
Yimeng Zhang ◽  
Yue Xu ◽  
...  
Keyword(s):  
Visual Cortex ◽  
Primary Visual Cortex ◽  
Large Scale ◽  
Complex Pattern ◽  
Two Photon ◽  
Photon Imaging ◽  
Two Photon Imaging

Multispot two-photon imaging of calcium waves dynamics in cardial tissue at 16Hz frame rate

2013 ◽  
Author(s):  
C. de Mauro ◽  
C. A. Cecchetti ◽  
D. Alfieri ◽  
G. Borile ◽  
A. Urbani ◽  
...  
Keyword(s):  
Frame Rate ◽  
Calcium Waves ◽  
Two Photon ◽  
Photon Imaging ◽  
Two Photon Imaging

scholarly journals Large-Scale 3D Two-Photon Imaging of Molecularly Identified CA1 Interneuron Dynamics in Behaving Mice

Neuron ◽  
2020 ◽  
Vol 108 (5) ◽  
pp. 968-983.e9
Author(s):  
Tristan Geiller ◽  
Bert Vancura ◽  
Satoshi Terada ◽  
Eirini Troullinou ◽  
Spyridon Chavlis ◽  
...  
Keyword(s):  
Large Scale ◽  
Two Photon ◽  
Photon Imaging ◽  
Two Photon Imaging

scholarly journals Two-photon imaging in mice shows striosomes and matrix have overlapping but differential reinforcement-related responses

eLife ◽  
2017 ◽  
Vol 6 ◽  
Author(s):  
Bernard Bloem ◽  
Rafiq Huda ◽  
Mriganka Sur ◽  
Ann M Graybiel
Keyword(s):  
Calcium Imaging ◽  
In Vivo Imaging ◽  
Differential Reinforcement ◽  
Response Strength ◽  
Two Photon ◽  
Expected Outcome ◽  
Photon Imaging ◽  
Two Photon Imaging ◽  
Circuit Function

Striosomes were discovered several decades ago as neurochemically identified zones in the striatum, yet technical hurdles have hampered the study of the functions of these striatal compartments. Here we used 2-photon calcium imaging in neuronal birthdate-labeled Mash1-CreER;Ai14 mice to image simultaneously the activity of striosomal and matrix neurons as mice performed an auditory conditioning task. With this method, we identified circumscribed zones of tdTomato-labeled neuropil that correspond to striosomes as verified immunohistochemically. Neurons in both striosomes and matrix responded to reward-predicting cues and were active during or after consummatory licking. However, we found quantitative differences in response strength: striosomal neurons fired more to reward-predicting cues and encoded more information about expected outcome as mice learned the task, whereas matrix neurons were more strongly modulated by recent reward history. These findings open the possibility of harnessing in vivo imaging to determine the contributions of striosomes and matrix to striatal circuit function.

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