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Brain-wide 3D imaging of neuronal activity in Caenorhabditis elegans with sculpted light

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TLDR
A two-photon technique for brain-wide calcium imaging in Caenorhabditis elegans, using wide-field temporal focusing (WF-TeFo) and a nuclear-localized, genetically encoded calcium indicator, NLS-GCaMP5K, that permits unambiguous discrimination of individual neurons within the densely packed head ganglia of C. elegans.
Abstract
Recent efforts in neuroscience research have been aimed at obtaining detailed anatomical neuronal wiring maps as well as information on how neurons in these networks engage in dynamic activities. Although the entire connectivity map of the nervous system of Caenorhabditis elegans has been known for more than 25 years, this knowledge has not been sufficient to predict all functional connections underlying behavior. To approach this goal, we developed a two-photon technique for brain-wide calcium imaging in C. elegans, using wide-field temporal focusing (WF-TeFo). Pivotal to our results was the use of a nuclear-localized, genetically encoded calcium indicator, NLS-GCaMP5K, that permits unambiguous discrimination of individual neurons within the densely packed head ganglia of C. elegans. We demonstrate near-simultaneous recording of activity of up to 70% of all head neurons. In combination with a lab-on-a-chip device for stimulus delivery, this method provides an enabling platform for establishing functional maps of neuronal networks.

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Citations
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Dual-color multiple-particle tracking at 50-nm localization and over 100-µm range in 3D with temporal focusing two-photon microscopy.

TL;DR: This method provides a simple wide-field fluorescence imaging approach for deep multiple-particle tracking and demonstrates its capability of tracking fast swimming microbes at speed of ~200 µm/s.
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Deep-Learning-Based Virtual Refocusing of Images Using an Engineered Point-Spread Function

TL;DR: In this article, a virtual image refocusing method over an extended depth of field (DOF) enabled by cascaded neural networks and a double-helix point spread function (DH-PSF) is presented.
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Real-time denoising enables high-sensitivity fluorescence time-lapse imaging beyond the shot-noise limit

TL;DR: DeepCAD-RT as mentioned in this paper is a self-supervised deep learning method for real-time noise suppression in fluorescence microscopy, which reduces the number of network parameters by 94%, memory consumption by 27-fold and processing time by a factor of 20.
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A double-sided microscope to realize whole-ganglion imaging of membrane potential in the medicinal leech.

TL;DR: A voltage-sensitive dye (VSD) imaging technique based on a double-sided microscope that can image two sides of a nervous system simultaneously was developed and applied to the segmental ganglia of the medicinal leech.
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Learning to Reconstruct Confocal Microscopy Stacks From Single Light Field Images

TL;DR: In this article, a fully convolutional neural network architecture inspired by the U-Net design was proposed to reconstruct confocal microscopy stacks from single light field images. But this method is not suitable for imaging highly dynamic and light sensitive events, and in closed-loop systems, where latency due to the reconstruction time is crucial.
References
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Journal ArticleDOI

The structure of the nervous system of the nematode Caenorhabditis elegans

TL;DR: The structure and connectivity of the nervous system of the nematode Caenorhabditis elegans has been deduced from reconstructions of electron micrographs of serial sections as discussed by the authors.
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Ultrasensitive fluorescent proteins for imaging neuronal activity.

TL;DR: A family of ultrasensitive protein calcium sensors (GCaMP6) that outperformed other sensors in cultured neurons and in zebrafish, flies and mice in vivo are developed and provide new windows into the organization and dynamics of neural circuits over multiple spatial and temporal scales.
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Functional network organization of the human brain

TL;DR: In this article, the authors studied functional brain organization in healthy adults using resting state functional connectivity MRI and proposed two novel brain wide graphs, one of 264 putative functional areas, the other a modification of voxelwise networks that eliminates potentially artificial short-distance relationships.
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Measurement of current-voltage relations in the membrane of the giant axon of Loligo.

TL;DR: The importance of ionic movements in excitable tissues has been emphasized by a number of recent experiments which are consistent with the theory that nervous conduction depends on a specific increase in permeability which allows sodium ions to move from the more concentrated solution outside a nerve fibre to the more dilute solution inside it.
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Reconstruction of zebrafish early embryonic development by scanned light sheet microscopy.

TL;DR: This work developed digital scanned laser light sheet fluorescence microscopy and recorded nuclei localization and movement in entire wild-type and mutant zebrafish embryos over the first 24 hours of development to derive a model of germ layer formation and show that the mesendoderm forms from one-third of the embryo's cells in a single event.
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