Miniscope GRIN Lens System for Calcium Imaging of Neuronal Activity from Deep Brain Structures in Behaving Animals

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Abstract

Visualizing neural activity from deep brain regions in freely behaving animals through miniature fluorescent microscope (miniscope) systems is becoming more important for understanding neural encoding mechanisms underlying cognitive functions. Here we present our custom-designed miniscope GRadient INdex (GRIN) lens system that enables simultaneously recording from hundreds of neurons for months. This article includes miniscope design, the surgical procedure for GRIN lens implantation, miniscope mounting on the head of a mouse, and data acquisition and analysis. First, a target brain region is labeled with virus expressing GCaMP6; second, a GRIN lens is implanted above the target brain region; third, following mouse surgical recovery, a miniscope is mounted on the head of the mouse above the GRIN lens; and finally, neural activity is recorded from the freely behaving mouse. This system can be applied to recording the same population of neurons longitudinally, enabling the elucidation of neural mechanisms underlying behavioral control. © 2018 by John Wiley & Sons, Inc.

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Most cited references 15

Record: found
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Article: not found

Fluorescent indicators for Ca2+ based on green fluorescent proteins and calmodulin.

A Miyawaki, J Llopis, R. Heim … (1997)

Important Ca2+ signals in the cytosol and organelles are often extremely localized and hard to measure. To overcome this problem we have constructed new fluorescent indicators for Ca2+ that are genetically encoded without cofactors and are targetable to specific intracellular locations. We have dubbed these fluorescent indicators 'cameleons'. They consist of tandem fusions of a blue- or cyan-emitting mutant of the green fluorescent protein (GFP), calmodulin, the calmodulin-binding peptide M13, and an enhanced green- or yellow-emitting GFP. Binding of Ca2+ makes calmodulin wrap around the M13 domain, increasing the fluorescence resonance energy transfer (FRET) between the flanking GFPs. Calmodulin mutations can tune the Ca2+ affinities to measure free Ca2+ concentrations in the range 10(-8) to 10(-2) M. We have visualized free Ca2+ dynamics in the cytosol, nucleus and endoplasmic reticulum of single HeLa cells transfected with complementary DNAs encoding chimaeras bearing appropriate localization signals. Ca2+ concentration in the endoplasmic reticulum of individual cells ranged from 60 to 400 microM at rest, and 1 to 50 microM after Ca2+ mobilization. FRET is also an indicator of the reversible intermolecular association of cyan-GFP-labelled calmodulin with yellow-GFP-labelled M13. Thus FRET between GFP mutants can monitor localized Ca2+ signals and protein heterodimerization in individual live cells.

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Record: found
Abstract: not found
Article: not found

Two-photon laser scanning fluorescence microscopy

W Denk, J G Strickler, W. Webb (1990)

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Record: found
Abstract: found
Article: not found

Optimization of a GCaMP calcium indicator for neural activity imaging.

Jasper Akerboom, Tsai-Wen Chen, Trevor J Wardill … (2012)

Genetically encoded calcium indicators (GECIs) are powerful tools for systems neuroscience. Recent efforts in protein engineering have significantly increased the performance of GECIs. The state-of-the art single-wavelength GECI, GCaMP3, has been deployed in a number of model organisms and can reliably detect three or more action potentials in short bursts in several systems in vivo. Through protein structure determination, targeted mutagenesis, high-throughput screening, and a battery of in vitro assays, we have increased the dynamic range of GCaMP3 by severalfold, creating a family of "GCaMP5" sensors. We tested GCaMP5s in several systems: cultured neurons and astrocytes, mouse retina, and in vivo in Caenorhabditis chemosensory neurons, Drosophila larval neuromuscular junction and adult antennal lobe, zebrafish retina and tectum, and mouse visual cortex. Signal-to-noise ratio was improved by at least 2- to 3-fold. In the visual cortex, two GCaMP5 variants detected twice as many visual stimulus-responsive cells as GCaMP3. By combining in vivo imaging with electrophysiology we show that GCaMP5 fluorescence provides a more reliable measure of neuronal activity than its predecessor GCaMP3. GCaMP5 allows more sensitive detection of neural activity in vivo and may find widespread applications for cellular imaging in general.

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Author and article information

Journal

Title: Current Protocols in Neuroscience

Abbreviated Title: Current Protocols in Neuroscience

Publisher: Wiley

ISSN: 19348584

Publication date Created: January 2019

Publication date (Print): January 2019

Publication date (Electronic): October 13 2018

Volume: 86

Issue: 1

Page: e56

Affiliations

[1 ]Intramural Research Program, National Institute on Drug Abuse, National Institutes of Health; Baltimore Maryland

[2 ]Transgenic Section, Laboratory of Neurogenetics, National Institute of Aging, National Institutes of Health; Bethesda Maryland

[3 ]Hefei National Laboratory for Physical Sciences at the Microscale, School of Life Sciences, University of Science and Technology of China; Hefei China

[4 ]Department of Zoology and Physiology, University of Wyoming College of Arts and Sciences; Laramie Wyoming

[5 ]The Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine; Baltimore Maryland