High-Speed Two-Photon Calcium Imaging of Neuronal Population Activity Using Acousto-Optic Deflectors

2014 ◽  
Vol 2014 (6) ◽  
pp. pdb.prot081778-pdb.prot081778 ◽  
Author(s):  
B. F. Grewe ◽  
F. Helmchen
Keyword(s):  
Calcium Imaging ◽  
High Speed ◽  
Neuronal Population ◽  
Population Activity ◽  
Two Photon

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.

scholarly journals Diesel2p mesoscope with dual independent scan engines for flexible capture of dynamics in distributed neural circuitry

2020 ◽  
Author(s):  
Che-Hang Yu ◽  
Jeffrey N. Stirman ◽  
Yiyi Yu ◽  
Riichiro Hira ◽  
Spencer L. Smith
Keyword(s):  
Adaptive Optics ◽  
Calcium Imaging ◽  
High Speed ◽  
Optical Design ◽  
Neural Circuitry ◽  
Brain Regions ◽  
Field Of View ◽  
Large Field ◽  
Two Photon ◽  
Subcellular Resolution

AbstractImaging the activity of neurons that are widely distributed across brain regions deep in scattering tissue at high speed remains challenging. Here, we introduce an open-source system with Dual Independent Enhanced Scan Engines for Large Field-of-view Two-Photon imaging (Diesel2p). Combining novel optical design, adaptive optics, and temporal multiplexing, the system offers subcellular resolution over a large field-of-view (∼ 25 mm2) with independent scan engines. We demonstrate the flexibility and various use cases of this system for calcium imaging of neurons in the living brain.

scholarly journals A computational toolbox and step-by-step tutorial for the analysis of neuronal population dynamics in calcium imaging data

2017 ◽  
Author(s):  
Sebastián A. Romano ◽  
Verónica Pérez-Schuster ◽  
Adrien Jouary ◽  
Alessia Candeo ◽  
Jonathan Boulanger-Weill ◽  
...  
Keyword(s):  
Population Dynamics ◽  
Calcium Imaging ◽  
Single Neuron ◽  
Neuronal Population ◽  
Population Coding ◽  
Imaging Data ◽  
Neuronal Responses ◽  
Population Activity ◽  
The Brain

The development of new imaging and optogenetics techniques to study the dynamics of large neuronal circuits is generating datasets of unprecedented volume and complexity, demanding the development of appropriate analysis tools. We present a tutorial for the use of a comprehensive computational toolbox for the analysis of neuronal population activity imaging. It consists of tools for image pre-processing and segmentation, estimation of significant single-neuron single-trial signals, mapping event-related neuronal responses, detection of activity-correlated neuronal clusters, exploration of population dynamics, and analysis of clusters’ features against surrogate control datasets. They are integrated in a modular and versatile processing pipeline, adaptable to different needs. The clustering module is capable of detecting flexible, dynamically activated neuronal assemblies, consistent with the distributed population coding of the brain. We demonstrate the suitability of the toolbox for a variety of calcium imaging datasets, and provide a case study to explain its implementation.

scholarly journals High-speed, multi-Z confocal microscopy for voltage imaging in densely labeled neuronal populations

2021 ◽  
Author(s):  
Timothy D Weber ◽  
Maria V Moya ◽  
Jerome Mertz ◽  
Michael N Economo
Keyword(s):  
High Speed ◽  
Signal To Noise Ratio ◽  
Neuronal Population ◽  
Great Promise ◽  
High Signal ◽  
Signal To Noise ◽  
Population Activity ◽  
Voltage Imaging ◽  
Neuronal Populations ◽  
Optical Crosstalk

Genetically encoded voltage indicators (GEVIs) hold great promise for monitoring neuronal population activity, but GEVI imaging in dense neuronal populations remains difficult due to a lack of contrast and/or speed. To address this challenge, we developed a novel confocal microscope that allows simultaneous multiplane imaging with high-contrast at near-kHz rates. This approach enables high signal-to-noise ratio voltage imaging in densely labeled populations and minimizes optical crosstalk during concurrent optogenetic photostimulation.

scholarly journals Diesel2p mesoscope with dual independent scan engines for flexible capture of dynamics in distributed neural circuitry

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Che-Hang Yu ◽  
Jeffrey N. Stirman ◽  
Yiyi Yu ◽  
Riichiro Hira ◽  
Spencer L. Smith
Keyword(s):  
Adaptive Optics ◽  
Calcium Imaging ◽  
High Speed ◽  
Optical Design ◽  
Neural Circuitry ◽  
Brain Regions ◽  
Field Of View ◽  
Large Field ◽  
Two Photon ◽  
Subcellular Resolution

AbstractImaging the activity of neurons that are widely distributed across brain regions deep in scattering tissue at high speed remains challenging. Here, we introduce an open-source system with Dual Independent Enhanced Scan Engines for Large field-of-view Two-Photon imaging (Diesel2p). Combining optical design, adaptive optics, and temporal multiplexing, the system offers subcellular resolution over a large field-of-view of ~25 mm2, encompassing distances up to 7 mm, with independent scan engines. We demonstrate the flexibility and various use cases of this system for calcium imaging of neurons in the living 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 High-speed label-free two-photon fluorescence microscopy of metabolic transients during neuronal activity

2021 ◽  
Vol 118 (8) ◽  
pp. 081104
Author(s):  
Andrew J. Bower ◽  
Carlos Renteria ◽  
Joanne Li ◽  
Marina Marjanovic ◽  
Ronit Barkalifa ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Neuronal Activity ◽  
High Speed ◽  
Label Free ◽  
Two Photon ◽  
Metabolic Transients ◽  
Two Photon Fluorescence ◽  
Photon Fluorescence

scholarly journals Two-photon calcium imaging during fictive navigation in virtual environments

2013 ◽  
Vol 7 ◽  
Author(s):  
Misha B. Ahrens ◽  
Kuo Hua Huang ◽  
Sujatha Narayan ◽  
Brett D. Mensh ◽  
Florian Engert
Keyword(s):  
Virtual Environments ◽  
Calcium Imaging ◽  
Two Photon
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