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Deciphering spatio-temporal fluorescence changes using multi-threshold event detection (MTED)

View ORCID ProfileFranziska E. Müller, View ORCID ProfileVolodymyr Cherkas, Gebhard Stopper, View ORCID ProfileLaura C. Caudal, View ORCID ProfileLaura Stopper, View ORCID ProfileFrank Kirchhoff, Christian Henneberger, View ORCID ProfileEvgeni G. Ponimaskin, View ORCID ProfileAndre Zeug
doi: https://doi.org/10.1101/2020.12.06.413492
Franziska E. Müller
1Cellular Neurophysiology, Hannover Medical School, Hannover, Germany
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Volodymyr Cherkas
1Cellular Neurophysiology, Hannover Medical School, Hannover, Germany
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Gebhard Stopper
2Department of Molecular Physiology, Center for Integrative Physiology and Molecular Medicine (CIPMM), University of Saarland, Homburg, Germany
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Laura C. Caudal
2Department of Molecular Physiology, Center for Integrative Physiology and Molecular Medicine (CIPMM), University of Saarland, Homburg, Germany
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Laura Stopper
2Department of Molecular Physiology, Center for Integrative Physiology and Molecular Medicine (CIPMM), University of Saarland, Homburg, Germany
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Frank Kirchhoff
2Department of Molecular Physiology, Center for Integrative Physiology and Molecular Medicine (CIPMM), University of Saarland, Homburg, Germany
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Christian Henneberger
3Institute of Cellular Neurosciences, Medical Faculty, University of Bonn, Bonn, Germany
4German Center for Neurodegenerative Diseases (DZNE), Bonn, Germany
5Institute of Neurology, University College London, London, United Kingdom
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Evgeni G. Ponimaskin
1Cellular Neurophysiology, Hannover Medical School, Hannover, Germany
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  • For correspondence: zeug.andre@mh-hannover.de
Andre Zeug
1Cellular Neurophysiology, Hannover Medical School, Hannover, Germany
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  • For correspondence: zeug.andre@mh-hannover.de
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Abstract

Recent achievements in indicator optimization and imaging techniques promote the exploration of Ca2+ activity patterns as a main second messenger in many organs. Astrocytes are important regulators of brain activity and well known for their Ca2+-dependent modulation of neurons. However, standardized methods to analyze and interpret Ca2+ activity recordings are missing and hindering global comparisons. Here, we present a biophysics-based concept to analyze Ca2+signals, which includes multiple thresholds and provides the experimenter with a comprehensive toolbox for a differentiated and in-depth characterization of Ca2+ signals. We analyzed various ex vivo and in vivo imaging datasets and verify the validity of our multi-threshold event detection (MTED) algorithm across Ca2+ indicators, imaging setups, and model systems from primary cell culture to awake, head-fixed mice. Applying our MTED concept enables standardized analysis and advances research using optical readouts of cellular activity to decrypt brain function. It allowed us to obtain new insights into the complex dependence of Ca2+activity patterns on temperature and neuronal activity.

Highlights

  • → We present a robust pixel-based algorithm to analyze multidimensional fluorescence data.

  • → Automated multiple-threshold analysis accurately quantifies changes in fluorescence across magnitudes.

  • → It characterizes the complexity of dynamic and overlapping activity patterns of Ca2+ activity of astrocytes in vitro, in situ, and in vivo.

  • → Its application provides quantitative parameters how temperature and neuronal activity determine astrocytic Ca2+ activity.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted December 07, 2020.
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Deciphering spatio-temporal fluorescence changes using multi-threshold event detection (MTED)
Franziska E. Müller, Volodymyr Cherkas, Gebhard Stopper, Laura C. Caudal, Laura Stopper, Frank Kirchhoff, Christian Henneberger, Evgeni G. Ponimaskin, Andre Zeug
bioRxiv 2020.12.06.413492; doi: https://doi.org/10.1101/2020.12.06.413492
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Deciphering spatio-temporal fluorescence changes using multi-threshold event detection (MTED)
Franziska E. Müller, Volodymyr Cherkas, Gebhard Stopper, Laura C. Caudal, Laura Stopper, Frank Kirchhoff, Christian Henneberger, Evgeni G. Ponimaskin, Andre Zeug
bioRxiv 2020.12.06.413492; doi: https://doi.org/10.1101/2020.12.06.413492

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