Studying Axon-Astrocyte Functional Interactions by 3D Two-Photon Ca<sup>2+</sup> Imaging: A Practical Guide to Experiments and "Big Data" Analysis.

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State: Public
Version: Final published version
Serval ID
serval:BIB_B24650E8D046
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Studying Axon-Astrocyte Functional Interactions by 3D Two-Photon Ca<sup>2+</sup> Imaging: A Practical Guide to Experiments and "Big Data" Analysis.
Journal
Frontiers in Cellular Neuroscience
Author(s)
Savtchouk I., Carriero G., Volterra A.
ISSN
1662-5102 (Print)
ISSN-L
1662-5102
Publication state
Published
Issued date
2018
Peer-reviewed
Oui
Volume
12
Pages
98
Language
english
Abstract
Recent advances in fast volumetric imaging have enabled rapid generation of large amounts of multi-dimensional functional data. While many computer frameworks exist for data storage and analysis of the multi-gigabyte Ca <sup>2+</sup> imaging experiments in neurons, they are less useful for analyzing Ca <sup>2+</sup> dynamics in astrocytes, where transients do not follow a predictable spatio-temporal distribution pattern. In this manuscript, we provide a detailed protocol and commentary for recording and analyzing three-dimensional (3D) Ca <sup>2+</sup> transients through time in GCaMP6f-expressing astrocytes of adult brain slices in response to axonal stimulation, using our recently developed tools to perform interactive exploration, filtering, and time-correlation analysis of the transients. In addition to the protocol, we release our in-house software tools and discuss parameters pertinent to conducting axonal stimulation/response experiments across various brain regions and conditions. Our software tools are available from the Volterra Lab webpage at https://wwwfbm.unil.ch/dnf/group/glia-an-active-synaptic-partner/member/volterra-andrea-volterra in the form of software plugins for Image J (NIH)-a de facto standard in scientific image analysis. Three programs are available: <i>MultiROI_TZ_profiler</i> for interactive graphing of several movable ROIs simultaneously, <i>Gaussian_Filter5D</i> for Gaussian filtering in several dimensions, and <i>Correlation_Calculator</i> for computing various cross-correlation parameters on voxel collections through time.

Keywords
3D calcium imaging, 5D datasets, ImageJ plugin, VoI, astrocytes, axons, calcium signaling, volumetric imaging
Pubmed
Web of science
Open Access
Yes
Create date
03/05/2018 14:52
Last modification date
20/08/2019 15:21
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