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Relationship between simultaneously recorded spiking activity and fluorescence signal in GCaMP6 transgenic mice

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Cite this article as: eLife 2021;10:e51675 doi: 10.7554/eLife.51675

Abstract

Fluorescent calcium indicators are often used to investigate neural dynamics, but the relationship between fluorescence and action potentials (APs) remains unclear. Most APs can be detected when the soma almost fills the microscope's field of view, but calcium indicators are often used to image populations of neurons, necessitating a large field of view, generating fewer photons per neuron, and compromising AP detection. Here we characterized the AP-fluorescence transfer function in vivo for 48 layer 2/3 pyramidal neurons in primary visual cortex, with simultaneous calcium imaging and cell-attached recordings from transgenic mice expressing GCaMP6s or GCaMP6f. While most APs were detected under optimal conditions, under conditions typical of population imaging studies only a minority of 1AP and 2AP events were detected (often <10% and ~20-30%, respectively), emphasizing the limits of AP detection under more realistic imaging conditions.

Article and author information

Author details

  1. Lawrence Huang

    Electrophysiology, Allen Institute for Brain Science, Seattle, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Peter Ledochowitsch

    MindScope Program, Allen Institute for Brain Science, Seattle, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Ulf Knoblich

    Structured Science, Allen Institute for Brain Science, Seattle, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Jérôme Lecoq

    MindScope Program, Allen Institute for Brain Science, Seattle, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Gabe J Murphy

    Structured Science, Allen Institute for Brain Science, Seattle, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Clay Reid

    Structured Science, Allen Institute for Brain Science, Seattle, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8697-6797
  7. Saskia E J de Vries

    MindScope Program, Allen Institute for Brain Science, Seattle, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-3704-3499
  8. Christof Koch

    MindScope Program, Allen Institute for Brain Science, Seattle, United States
    Competing interests
    The authors declare that no competing interests exist.
  9. Hongkui Zeng

    Structured Science, Allen Institute for Brain Science, Seattle, United States
    For correspondence
    hongkuiz@alleninstitute.org
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0326-5878
  10. Michael A Buice

    MindScope Program, Allen Institute for Brain Science, Seattle, United States
    Competing interests
    The authors declare that no competing interests exist.
  11. Jack Waters

    Structured Science, Allen Institute for Brain Science, Seattle, United States
    For correspondence
    jackw@alleninstitute.org
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-2312-4183
  12. Lu Li

    Structured Science, Allen Institute for Brain Science, Seattle, United States
    For correspondence
    lilu67@mail.sysu.edu.cn
    Competing interests
    The authors declare that no competing interests exist.

Funding

Allen Institute for Brain Science (program funds)

  • Lawrence Huang
  • Peter Ledochowitsch
  • Ulf Knoblich
  • Jérôme Lecoq
  • Gabe J Murphy
  • Clay Reid
  • Saskia E J de Vries
  • Christof Koch
  • Hongkui Zeng
  • Michael A Buice
  • Jack Waters
  • Lu Li

National Natural Science Foundation of China (NSFC31871055)

  • Lu Li

Guangdong Science and Technology Department (2017B030314026)

  • Lu Li

This work is funded by the Allen Institute for Brain Science. The funder had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: Experimental procedures were conducted in accordance with NIH guidelines and approved by the Institutional Animal Care and Use Committee (IACUC) of the Allen Institute for Brain Science under protocol number 1509.

Reviewing Editor

  1. Gary L Westbrook, Oregon Health and Science University, United States

Publication history

  1. Received: September 6, 2019
  2. Accepted: March 5, 2021
  3. Accepted Manuscript published: March 8, 2021 (version 1)

Copyright

© 2021, Huang et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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