scholarly journals Parallel holographic illumination enables sub-millisecond two-photon optogenetic activation in mouse visual cortex in vivo

2018 ◽  
Author(s):  
I-Wen Chen ◽  
Emiliano Ronzitti ◽  
Brian R. Lee ◽  
Tanya L. Daigle ◽  
Hongkui Zeng ◽  
...  
Keyword(s):  
Visual Cortex ◽  
Temporal Resolution ◽  
Optical Control ◽  
Target Cells ◽  
High Temporal Resolution ◽  
Two Photon ◽  
Brain Functions ◽  
All Optical ◽  
Mouse Visual Cortex

AbstractSelective control of action potential generation in individual cells from a neuronal ensemble is desirable for dissecting circuit mechanisms underlying perception and behavior. Here, by using two-photon (2P) temporally focused computer-generated holography (TF-CGH), we demonstrate optical manipulation of neuronal excitability at the supragranular layers of anesthetized mouse visual cortex. Utilizing amplified laser-pulses delivered via a localized holographic spot, our optical system achieves suprathreshold activation by exciting either of the three optogenetic actuators, ReaChR, CoChR or ChrimsonR, with brief illumination (≤ 10 ms) at moderate excitation power ((in average ≤ 0.2 mW/µm2 corresponding to ≤ 25 mW/cell). Using 2P-guided whole-cell or cell-attached recordings in positive neurons expressing respective opsin in vivo, we find that parallel illumination induces spikes of millisecond temporal resolution and sub-millisecond precision, which are preserved upon repetitive illuminations up to tens of Hz. Holographic stimulation thus enables temporally precise optogenetic activation independently of opsin’s channel kinetics. Furthermore, we demonstrate that parallel optogenetic activation can be combined with functional imaging for all-optical control of a neuronal sub-population that co-expresses the photosensitive opsin ReaChR and the calcium indicator GCaMP6s. Parallel optical control of neuronal activity with cellular resolution and millisecond temporal precision should be advantageous for investigating neuronal connections and further yielding causal links between connectivity, microcircuit dynamics, and brain functions.Significance statementRecent development of optogenetics allows probing the neuronal microcircuit with light by optically actuating genetically-encoded light-sensitive opsins expressed in the target cells. Here, we apply holographic light shaping and temporal focusing to simultaneously deliver axially-confined holographic patterns to opsin-positive cells situated in the living mouse cortex. Parallel illumination efficiently induces action potentials with high temporal resolution and precision for three opsins of different kinetics. We demonstrated all-optical experiments by extending the parallel optogenetic activation at low intensity to multiple neurons and concurrently monitoring their calcium dynamics. These results demonstrate fast and temporally precise in vivo control of a neuronal sub-population, opening new opportunities to reveal circuit mechanisms underlying brain functions.

scholarly journals Navigating the translational roadblock: Towards highly specific and effective all-optical interrogations of neural circuits

2020 ◽  
Author(s):  
Ting Fu ◽  
Isabelle Arnoux ◽  
Jan Döring ◽  
Hirofumi Watari ◽  
Ignas Stasevicius ◽  
...  
Keyword(s):  
Visual Cortex ◽  
Calcium Imaging ◽  
Cortical Neurons ◽  
Activity Patterns ◽  
Detection Algorithm ◽  
Two Photon ◽  
All Optical ◽  
Laser Sources ◽  
Mouse Visual Cortex

AbstractTwo-photon (2-P) all-optical approaches combine in vivo 2-P calcium imaging and 2-P optogenetic modulations and have the potential to build a framework for network-based therapies, e.g. for rebalancing maladaptive activity patterns in preclinical models of neurological disorders. Here, our goal was to tailor these approaches for this purpose: Firstly, we combined in vivo juxtacellular recordings and GCaMP6f-based 2-P calcium imaging in layer II/III of mouse visual cortex to tune our detection algorithm towards a 100 % specific identification of AP-related calcium transients. False-positive-free detection was achieved at a sensitivity of approximately 73 %. To further increase specificity, secondly, we minimized photostimulation artifacts as a potential source for false-positives by using extended-wavelength-spectrum laser sources for optogenetic stimulation of the excitatory opsin C1V1. We achieved artifact-free all-optical experiments performing photostimulations at 1100 nm or higher and simultaneous calcium imaging at 920 nm in mouse visual cortex in vivo. Thirdly, we determined the spectral range for maximizing efficacy of optogenetic control by performing 2-P photostimulations of individual neurons with wavelengths up to 1300 nm. The rate of evoked transients in GCaMP6f/C1V1-co-expressing cortical neurons peaked already at 1100 nm. By refining spike detection and defining 1100 nm as the optimal wavelength for artifact-free and effective stimulations of C1V1 in GCaMP-based all-optical interrogations, we increased the translational value of these approaches, e.g. for the use in preclinical applications of network-based therapies.One Sentence SummaryWe maximize translational relevance of 2-P all-optical physiology by increasing specificity, minimizing artifacts and optimizing stimulation efficacy.

scholarly journals In Vivo Two-Photon Ca2+ Imaging Reveals Selective Reward Effects on Stimulus-Specific Assemblies in Mouse Visual Cortex

2013 ◽  
Vol 33 (28) ◽  
pp. 11540-11555 ◽  
Author(s):  
P. M. Goltstein ◽  
E. B. J. Coffey ◽  
P. R. Roelfsema ◽  
C. M. A. Pennartz
Keyword(s):  
Visual Cortex ◽  
Two Photon ◽  
Mouse Visual Cortex ◽  
Reward Effects

In vivo two-photon imaging analysis of development of direction and orientation selectivity in the mouse visual cortex

2011 ◽  
Vol 71 ◽  
pp. e257
Author(s):  
Madoka Narushima ◽  
Nathalie L. Rochefort ◽  
Christine Grienberger ◽  
Nima Marandi ◽  
Arthur Konnerth
Keyword(s):  
Visual Cortex ◽  
Orientation Selectivity ◽  
Imaging Analysis ◽  
Two Photon ◽  
Photon Imaging ◽  
Two Photon Imaging ◽  
Mouse Visual Cortex

scholarly journals Long-Range Interhemispheric Projection Neurons Show Biased Response Properties and Fine-Scale Local Subnetworks in Mouse Visual Cortex

2020 ◽  
Author(s):  
Kenta M Hagihara ◽  
Ayako Wendy Ishikawa ◽  
Yumiko Yoshimura ◽  
Yoshiaki Tagawa ◽  
Kenichi Ohki
Keyword(s):  
Visual Cortex ◽  
Long Range ◽  
Projection Neurons ◽  
Fine Scale ◽  
Local Connectivity ◽  
Cortical Projection ◽  
Response Properties ◽  
Brain Functions ◽  
Mouse Visual Cortex

Abstract Integration of information processed separately in distributed brain regions is essential for brain functions. This integration is enabled by long-range projection neurons, and further, concerted interactions between long-range projections and local microcircuits are crucial. It is not well known, however, how this interaction is implemented in cortical circuits. Here, to decipher this logic, using callosal projection neurons (CPNs) in layer 2/3 of the mouse visual cortex as a model of long-range projections, we found that CPNs exhibited distinct response properties and fine-scale local connectivity patterns. In vivo 2-photon calcium imaging revealed that CPNs showed a higher ipsilateral (to their somata) eye preference, and that CPN pairs showed stronger signal/noise correlation than random pairs. Slice recordings showed CPNs were preferentially connected to CPNs, demonstrating the existence of projection target-dependent fine-scale subnetworks. Collectively, our results suggest that long-range projection target predicts response properties and local connectivity of cortical projection neurons.

scholarly journals Astrocytes in the mouse visual cortex reliably respond to visual stimulation

2018 ◽  
Author(s):  
Keita Sonoda ◽  
Teppei Matsui ◽  
Haruhiko Bito ◽  
Kenichi Ohki
Keyword(s):  
Visual Cortex ◽  
Primary Visual Cortex ◽  
Common Mechanism ◽  
List Type ◽  
Visual Responses ◽  
Two Photon ◽  
Dual Color ◽  
Cross Correlation Analysis ◽  
Mouse Visual Cortex

AbstractAstrocytes are known to contact with a great number of synapses and may integrate sensory inputs. In the ferret primary visual cortex, astrocytes respond to a visual stimulus with a delay of several seconds with respect to the surrounding neurons. However, in the mouse visual cortex, it remains unclear whether astrocytes respond to visual stimulations. In this study, using dual-color simultaneous in vivo two-photon Ca2+ imaging of neurons and astrocytes in the awake mouse visual cortex, we examined the visual responsiveness of astrocytes and their precise response timing relative to the surrounding neurons. Neurons reliably responded to visual stimulations, whereas astrocytes often showed neuromodulator-mediated global activities, which largely masked small periodic activities. Administration of the selective α1-adrenergic receptor antagonist prazosin substantially reduced such global astrocytic activities without affecting the neuronal visual responses. In the presence of prazosin, astrocytes showed weak but consistent visual responses mostly at their somata. Cross-correlation analysis estimated that the astrocytic visual responses were delayed by approximately 5 s relative to the surrounding neuronal responses. In conclusion, our research demonstrated that astrocytes in the primary visual cortex of awake mice responded to visual stimuli with a delay of several seconds relative to the surrounding neurons, which may indicate the existence of a common mechanism of neuron–astrocyte communication across species.HighlightsWe performed dual-color in vivo two-photon Ca2+ imaging of neurons and astrocytes.α1-adrenoblocker prazosin substantially reduced global astrocytic activities.Astrocytes showed weak but reliable visual responses in the awake mouse visual cortex.Astrocytic visual responses were delayed by 5 s relative to the neuronal ones.

scholarly journals Simultaneous two-photon imaging and two-photon optogenetics of cortical circuits in three dimensions

eLife ◽  
2018 ◽  
Vol 7 ◽  
Author(s):  
Weijian Yang ◽  
Luis Carrillo-Reid ◽  
Yuki Bando ◽  
Darcy S Peterka ◽  
Rafael Yuste
Keyword(s):  
Neural Activity ◽  
Pyramidal Neurons ◽  
Three Dimensions ◽  
Cortical Circuits ◽  
Two Photon ◽  
Simultaneous Imaging ◽  
All Optical ◽  
Holographic Approach ◽  
Mouse Visual Cortex

The simultaneous imaging and manipulating of neural activity could enable the functional dissection of neural circuits. Here we have combined two-photon optogenetics with simultaneous volumetric two-photon calcium imaging to measure and manipulate neural activity in mouse neocortex in vivo in three-dimensions (3D) with cellular resolution. Using a hybrid holographic approach, we simultaneously photostimulate more than 80 neurons over 150 μm in depth in layer 2/3 of the mouse visual cortex, while simultaneously imaging the activity of the surrounding neurons. We validate the usefulness of the method by photoactivating in 3D selected groups of interneurons, suppressing the response of nearby pyramidal neurons to visual stimuli in awake animals. Our all-optical approach could be used as a general platform to read and write neuronal activity.

scholarly journals Multimodal in vivo recording using transparent graphene microelectrodes illuminates spatiotemporal seizure dynamics at the microscale

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Nicolette Driscoll ◽  
Richard E. Rosch ◽  
Brendan B. Murphy ◽  
Arian Ashourvan ◽  
Ramya Vishnubhotla ◽  
...  
Keyword(s):  
Temporal Resolution ◽  
Spatial Scales ◽  
Epileptic Seizures ◽  
Therapeutic Interventions ◽  
State Transitions ◽  
Integrated Analysis ◽  
High Temporal Resolution ◽  
Seizure Onset ◽  
Acute Seizures

AbstractNeurological disorders such as epilepsy arise from disrupted brain networks. Our capacity to treat these disorders is limited by our inability to map these networks at sufficient temporal and spatial scales to target interventions. Current best techniques either sample broad areas at low temporal resolution (e.g. calcium imaging) or record from discrete regions at high temporal resolution (e.g. electrophysiology). This limitation hampers our ability to understand and intervene in aberrations of network dynamics. Here we present a technique to map the onset and spatiotemporal spread of acute epileptic seizures in vivo by simultaneously recording high bandwidth microelectrocorticography and calcium fluorescence using transparent graphene microelectrode arrays. We integrate dynamic data features from both modalities using non-negative matrix factorization to identify sequential spatiotemporal patterns of seizure onset and evolution, revealing how the temporal progression of ictal electrophysiology is linked to the spatial evolution of the recruited seizure core. This integrated analysis of multimodal data reveals otherwise hidden state transitions in the spatial and temporal progression of acute seizures. The techniques demonstrated here may enable future targeted therapeutic interventions and novel spatially embedded models of local circuit dynamics during seizure onset and evolution.

scholarly journals In vivo magnetic particle imaging: angiography of inferior vena cava and aorta in rats using newly developed multicore particles

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Azadeh Mohtashamdolatshahi ◽  
Harald Kratz ◽  
Olaf Kosch ◽  
Ralf Hauptmann ◽  
Nicola Stolzenburg ◽  
...  
Keyword(s):  
Image Quality ◽  
Inferior Vena Cava ◽  
Temporal Resolution ◽  
Magnetic Particle ◽  
Inferior Vena ◽  
Vena Cava ◽  
High Temporal Resolution ◽  
Magnetic Particle Imaging ◽  
Particle Imaging

Abstract Magnetic Particle Imaging (MPI) is a new imaging modality, which maps the distribution of magnetic nanoparticles (MNP) in 3D with high temporal resolution. It thus may be suited for cardiovascular imaging. Its sensitivity and spatial resolution critically depend on the magnetic properties of MNP. Therefore, we used novel multicore nanoparticles (MCP 3) for in-vivo MPI in rats and analyzed dose requirements, sensitivity and detail resolution. 8 rats were examined using a preclinical MPI scanner (Bruker Biospin GmbH, Germany) equipped with a separate receive coil. MCP 3 and Resovist were administered intravenously (i.v.) into the rats’ tail veins at doses of 0.1, 0.05 and 0.025 mmol Fe/kg followed by serial MPI acquisition with a temporal resolution of 46 volumes per second. Based on a qualitative visual scoring system MCP 3–MPI images showed a significantly (P ≤ 0.05) higher image quality than Resovist-MPI images. Morphological features such as vessel lumen diameters (DL) of the inferior vena cava (IVC) and abdominal aorta (AA) could be assessed along a 2-cm segment in mesenteric area only after administration of MCP 3 at dosages of 0.1, 0.05 mmol Fe/kg. The mean DL ± SD estimated was 2.7 ± 0.6 mm for IVC and 2.4 ± 0.7 mm for AA. Evaluation of DL of the IVC and AA was not possible in Resovist-MPI images. Our results show, that MCP 3 provide better image quality at a lower dosage than Resovist. MCP 3-MPI with a clinically acceptable dose of 0.05 mmol Fe/kg increased the visibility of vessel lumens compared to Resovist-based MPI towards possible detection of vascular abnormalities such as stenosis or aneurysms, in vivo.

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