scholarly journals Simultaneous high-speed imaging and optogenetic inhibition in the intact mouse brain

2017 ◽  
Vol 7 (1) ◽  
Author(s):  
Serena Bovetti ◽  
Claudio Moretti ◽  
Stefano Zucca ◽  
Marco Dal Maschio ◽  
Paolo Bonifazi ◽  
...  
Keyword(s):  
Mouse Brain ◽  
High Speed ◽  
Pyramidal Neurons ◽  
Functional Organization ◽  
Single Photon ◽  
Electrophysiological Recording ◽  
Mammalian Brain ◽  
Cellular Mechanisms ◽  
High Speed Imaging ◽  
Intact Mouse

Abstract Genetically encoded calcium indicators and optogenetic actuators can report and manipulate the activity of specific neuronal populations. However, applying imaging and optogenetics simultaneously has been difficult to establish in the mammalian brain, even though combining the techniques would provide a powerful approach to reveal the functional organization of neural circuits. Here, we developed a technique based on patterned two-photon illumination to allow fast scanless imaging of GCaMP6 signals in the intact mouse brain at the same time as single-photon optogenetic inhibition with Archaerhodopsin. Using combined imaging and electrophysiological recording, we demonstrate that single and short bursts of action potentials in pyramidal neurons can be detected in the scanless modality at acquisition frequencies up to 1 kHz. Moreover, we demonstrate that our system strongly reduces the artifacts in the fluorescence detection that are induced by single-photon optogenetic illumination. Finally, we validated our technique investigating the role of parvalbumin-positive (PV) interneurons in the control of spontaneous cortical dynamics. Monitoring the activity of cellular populations on a precise spatiotemporal scale while manipulating neuronal activity with optogenetics provides a powerful tool to causally elucidate the cellular mechanisms underlying circuit function in the intact mammalian brain.

scholarly journals Erratum: Corrigendum: Simultaneous high-speed imaging and optogenetic inhibition in the intact mouse brain

2017 ◽  
Vol 7 (1) ◽  
Author(s):  
Serena Bovetti ◽  
Claudio Moretti ◽  
Stefano Zucca ◽  
Marco Dal Maschio ◽  
Paolo Bonifazi ◽  
...  
Keyword(s):  
Mouse Brain ◽  
High Speed ◽  
High Speed Imaging ◽  
Intact Mouse

scholarly journals Video-rate fluorescence lifetime imaging camera with CMOS single-photon avalanche diode arrays and high-speed imaging algorithm

2011 ◽  
Vol 16 (9) ◽  
pp. 096012 ◽  
Author(s):  
David D.-U. Li ◽  
Jochen Arlt ◽  
David Tyndall ◽  
Richard Walker ◽  
Justin Richardson ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
High Speed ◽  
Single Photon ◽  
Fluorescence Lifetime Imaging ◽  
High Speed Imaging ◽  
Imaging Algorithm ◽  
Avalanche Diode ◽  
Lifetime Imaging ◽  
Imaging Camera

scholarly journals Secretory vesicle trafficking in awake and anesthetized mice: differential speeds in axons versus synapses

2018 ◽  
Author(s):  
Johannes Knabbe ◽  
Joris Nassal ◽  
Matthijs Verhage ◽  
Thomas Kuner
Keyword(s):  
Transport Properties ◽  
Mouse Brain ◽  
Dense Core ◽  
Mammalian Brain ◽  
Brain State ◽  
Dense Core Vesicle ◽  
List Type ◽  
Intact Mouse ◽  
Synaptic Localization

AbstractNeuronal dense core vesicles (DCVs) transport many cargo molecules like neuropeptides and neurotrophins to their release sites in dendrites or axons. The transport properties of DCVs in axons of the intact mammalian brain are unknown. We used viral expression of a DCV cargo reporter (NPY-Venus/Cherry) in the thalamus and two-photon in vivo imaging to visualize axonal DCV trafficking in thalamo-cortical projections of anesthetized and awake mice. We found an average speed of 1 μm/s, maximal speeds of up to 5 μm/s and a pausing fraction of ~11%. Directionality of transport differed between anesthetized and awake mice. In vivo microtubule +-end extension imaging using Macf18-GFP revealed microtubular growth at 0.12 μm/s and provided positive identification of antero- and retrograde axonal transport. Consistent with previous reports, anterograde transport was faster (~2.1 μm/s) than retrograde transport (~1.4 μm/s). In summary, DCVs are transported with faster maximal speeds and lower pausing fraction in vivo compared to previous results obtained in vitro. Finally, we found that DCVs slowed down upon presynaptic bouton approach. We propose that this mechanism promotes synaptic localization and cargo release.Key pointsDespite their immense physiological and pathophysiological importance, we know very little about the biology of dense core vesicle (DCV) trafficking in the intact mammalian brain.DCVs are transported at similar average speeds in the anesthetized and awake mouse brain compared to neurons in culture, yet maximal speed and pausing fraction of transport were higher.Microtubule +-end extension imaging visualized microtubular growth at 0.12 μm/s and revealed that DCVs were transported faster in the anterograde direction.DCV transport slowed down upon presynaptic bouton approach, possibly promoting synaptic localization and cargo release.Our work provides a basis to extrapolate DCV transport properties determined in cultured neurons to the intact mouse brain and reveal novel features such as slowing upon bouton approach and brain state-dependent trafficking directionality.

scholarly journals Emergence of functional subnetworks in layer 2/3 cortex induced by sequential spikes in vivo

2016 ◽  
Vol 113 (10) ◽  
pp. E1372-E1381 ◽  
Author(s):  
Taekeun Kim ◽  
Won Chan Oh ◽  
Joon Ho Choi ◽  
Hyung-Bae Kwon
Keyword(s):  
Pyramidal Neurons ◽  
Sensory Information ◽  
Receptor Activation ◽  
Mammalian Brain ◽  
Time Interval ◽  
Cellular Mechanisms ◽  
Calcium Calmodulin Dependent ◽  
Layer 2 ◽  
Cortical Circuit

During cortical circuit development in the mammalian brain, groups of excitatory neurons that receive similar sensory information form microcircuits. However, cellular mechanisms underlying cortical microcircuit development remain poorly understood. Here we implemented combined two-photon imaging and photolysis in vivo to monitor and manipulate neuronal activities to study the processes underlying activity-dependent circuit changes. We found that repeated triggering of spike trains in a randomly chosen group of layer 2/3 pyramidal neurons in the somatosensory cortex triggered long-term plasticity of circuits (LTPc), resulting in the increased probability that the selected neurons would fire when action potentials of individual neurons in the group were evoked. Significant firing pattern changes were observed more frequently in the selected group of neurons than in neighboring control neurons, and the induction was dependent on the time interval between spikes, N-methyl-D-aspartate (NMDA) receptor activation, and Calcium/calmodulin-dependent protein kinase II (CaMKII) activation. In addition, LTPc was associated with an increase of activity from a portion of neighboring neurons with different probabilities. Thus, our results demonstrate that the formation of functional microcircuits requires broad network changes and that its directionality is nonrandom, which may be a general feature of cortical circuit assembly in the mammalian cortex.

Experimental Measurement of In-Plane Rolling Nonpneumatic Tire Vibrations Using High-Speed Imaging

2019 ◽  
Vol 47 (3) ◽  
pp. 196-210
Author(s):  
Meghashyam Panyam ◽  
Beshah Ayalew ◽  
Timothy Rhyne ◽  
Steve Cron ◽  
John Adcox
Keyword(s):  
High Speed ◽  
Image Correlation ◽  
Dynamic Mode Decomposition ◽  
Dynamic Mode ◽  
High Speed Imaging ◽  
Correlation Algorithm ◽  
Mode Decomposition ◽  
Series Of Experiments ◽  
Plane Deformations ◽  
Dominant Frequencies

ABSTRACT This article presents a novel experimental technique for measuring in-plane deformations and vibration modes of a rotating nonpneumatic tire subjected to obstacle impacts. The tire was mounted on a modified quarter-car test rig, which was built around one of the drums of a 500-horse power chassis dynamometer at Clemson University's International Center for Automotive Research. A series of experiments were conducted using a high-speed camera to capture the event of the rotating tire coming into contact with a cleat attached to the surface of the drum. The resulting video was processed using a two-dimensional digital image correlation algorithm to obtain in-plane radial and tangential deformation fields of the tire. The dynamic mode decomposition algorithm was implemented on the deformation fields to extract the dominant frequencies that were excited in the tire upon contact with the cleat. It was observed that the deformations and the modal frequencies estimated using this method were within a reasonable range of expected values. In general, the results indicate that the method used in this study can be a useful tool in measuring in-plane deformations of rolling tires without the need for additional sensors and wiring.

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