Understanding the neurovascular unit at multiple scales: Advantages and limitations of multi-photon and functional ultrasound imaging

Urban, Alan; Golgher, Lior; Brunner, Clément; Gdalyahu, Amos; Har-Gil, Hagai; Kain, David; Montaldo, Gabriel; Sironi, Laura; Blinder, Pablo

doi:10.1016/j.addr.2017.07.018

Cited by 43 publications

(55 citation statements)

References 338 publications

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“…ity, blood flow, etc.) (22). As rendering takes place off-line, experimental monitoring is done by routing one of the multiscaler outputs (SYNC, Figure 1a) to the analog-to-digital card of the existing system.…”

Section: Resultsmentioning

confidence: 99%

“…The histogram can either be rendered and viewed directly, or be processed further by registering it to the moving brain's frame of reference, and computing quantitative metrics about its content (neuronal activity, blood flow, etc.) (20). As rendering takes place off-line, experimental monitoring is done by routing one of the multiscaler outputs (SYNC, Figure 1a) to the analog-to-digital card of the existing system.…”

Section: Resultsmentioning

confidence: 99%

“…The neuroscience community is steadily striding towards high-throughput, rapid volumetric imaging of multiple brain regions (22,24). To achieve these feats, data acquisition hardware has to maintain single-photon sensitivity for tracking dim features, while still handling spouts of photons emitted from sparse bright features, this over lengthy imaging sessions.…”

Section: Discussionmentioning

confidence: 99%

“…Multi-photon laser scanning microscopy (MPLSM) provides a glimpse into the functioning mammalian brain with subcellular resolution (5,24,25). Recent improvements in optical microscope design, laser sources and fluorophores (22) have extended the use of MPLSM to challenging applications such as imaging of very large neuronal populations (21) and fast volumetric imaging (12). These applications face a common inherent limitation: a given rate of photon detections is spread across a rapidly increasing number of voxels sampled per second.…”

Section: Introductionmentioning

confidence: 99%

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PySight: plug and play photon counting for fast intravital microscopy

Har-Gil

Golgher

Israel

et al. 2018

Preprint

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Imaging increasingly large neuronal populations at high rates pushed multi-photon microscopy into the photon-deprived regime. We present PySight, an add-on hardware and software solution tailored for photon-deprived imaging conditions. PySight more than triples the median amplitude of neuronal calcium transients in awake mice, and facilitates single-trial intravital voltage imaging in fruit flies. Its unique data streaming architecture allowed us to image a fruit fly's olfactory response over 234×600×330µm 3 at 73 volumes per second, outperforming top-tier imaging setups while retaining over 200 times lower data rates. PySight requires no electronics expertise or custom synchronization boards, and its open-source software is extensible to any imaging method based on single-pixel (bucket) detectors. PySight offers an optimal data acquisition scheme for ever increasing imaging volumes of turbid living tissue.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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PySight: plug and play photon counting for fast intravital microscopy

Har-Gil

Golgher

Israel

et al. 2018

Preprint

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“…Functional ultrasound imaging (fUSI) is a method based on high-framerate plane-waves imaging to measure changes in cerebral blood volume (CBV) in response to neuronal activity (Mace et al, 2013;Mace E. et al, 2011;Montaldo et al, 2009;Urban et al, 2014aUrban et al, , 2017. fUSI has a small voxel size (~100x300x100 µm 3 ), an excellent temporal resolution (10 Hz framerate), a high depth of field (up to 1.5 cm), does not require contrast agents and is adaptable to freely-moving animals (Sieu et al, 2015;Tiran et al, 2017;Urban et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

A platform for brain-wide functional ultrasound imaging and analysis of circuit dynamics in behaving mice

Brunner

Grillet

Sans-Dublanc

et al. 2020

Preprint

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Imaging of large-scale circuit dynamics is crucial to gain a better understanding of brain function, but most techniques have a limited depth of field. Here we describe vfUSI, a platform for brain-wide volumetric functional ultrasound imaging of hemodynamic activity in awake head-fixed mice. We combined high-frequency 1024-channel 2D-array transducer with advanced multiplexing and highperformance computing for real-time 3D Power Doppler imaging at high spatiotemporal resolution (220x280x175-µm 3 voxel size, up to 6 Hz). In addition, we developed a standardized software pipeline for registration and segmentation based on the Allen Mouse Common Coordinate Framework, allowing for temporal analysis in 268 individual brain regions. We demonstrate the high sensitivity of vfUSI in multiple experimental situations where stimulus-evoked activity can be recorded using a minimal number of trials. We also mapped neural circuits in vivo across the whole brain during optogenetic activation of specific cell-types. Moreover, we revealed the sequential activation of sensory-motor regions during a grasping water droplet task. vfUSI will become a key neuroimaging technology because it combines ease of use, reliability, and affordability.

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NIR‐II Excitable Conjugated Polymer Dots with Bright NIR‐I Emission for Deep In Vivo Two‐Photon Brain Imaging Through Intact Skull

Wang

Liu

Feng

et al. 2019

Adv Funct Materials

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Methods for noninvasive brain imaging are highly desirable to study brain structures in neuroscience. Two‐photon fluorescence microscopy (2PFM) with near‐infrared (NIR) light excitation is a relatively noninvasive approach commonly used to study brain with high spatial resolution and large imaging depth. However, most of the current studies require cranial window implantation or skull‐thinning methods due to attenuation of excitation light. 2PFM through intact mouse skull is challenging due to strong scattering induced by skull bone. Herein, NIR‐II light excitable single‐chain conjugated polymer dots (CPdots) with bright fluorescence in NIR‐I region (peak at ≈725 nm and quantum yield of 20.6 ± 1.0%) are developed for deep in vivo two‐photon fluorescence (2PF) imaging of intact mouse brain. The synthesized CPdots exhibit good biocompatibility, high photostability, and large two‐photon absorption cross section. The CPdots allow 2PF images acquired upon excitation at 800, 1040 and 1200 nm with the highest signal‐to‐background ratio of 208 demonstrated for 1200 nm excitation. Moreover, a 3D reconstruction of the brain blood vessel network is obtained with a large vertical depth of 400 µm through intact skull. This work demonstrates great potential of bright NIR fluorophores for in vivo deep tissue imaging.

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Understanding the neurovascular unit at multiple scales: Advantages and limitations of multi-photon and functional ultrasound imaging

Cited by 43 publications

References 338 publications

PySight: plug and play photon counting for fast intravital microscopy

PySight: plug and play photon counting for fast intravital microscopy

A platform for brain-wide functional ultrasound imaging and analysis of circuit dynamics in behaving mice

NIR‐II Excitable Conjugated Polymer Dots with Bright NIR‐I Emission for Deep In Vivo Two‐Photon Brain Imaging Through Intact Skull

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