Cited by 1,967Advanced Fluorescence Microscopy TechniquesNeuroscience and Neuropharmacology ResearchIon Channels and Receptors

Imaging neural activity in worms, flies and mice with improved GCaMP calcium indicators

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Tian, Lin, Hires, S Andrew, Mao, Tianyi, Huber, Daniel, Chiappe, M Eugenia, Chalasani, Sreekanth H, Petreanu, Leopoldo, Akerboom, Jasper +6 more·2009·Nature Methods

Abstract

Genetically encoded calcium indicators (GECIs) can be used to image activity in defined neuronal populations. However, current GECIs produce inferior signals compared to synthetic indicators and recording electrodes, precluding detection of low firing rates. We developed a single-wavelength GCaMP2-based GECI (GCaMP3), with increased baseline fluorescence (3-fold), increased dynamic range (3-fold) and higher affinity for calcium (1.3-fold). We detected GCaMP3 fluorescence changes triggered by single action potentials in pyramidal cell dendrites, with signal-to-noise ratio and photostability substantially better than those of GCaMP2, D3cpVenus and TN-XXL. In Caenorhabditis elegans chemosensory neurons and the Drosophila melanogaster antennal lobe, sensory stimulation-evoked fluorescence responses were significantly enhanced with GCaMP3 (4-6-fold). In somatosensory and motor cortical neurons in the intact mouse, GCaMP3 detected calcium transients with amplitudes linearly dependent on action potential number. Long-term imaging in the motor cortex of behaving mice revealed large fluorescence changes in imaged neurons over months.

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  • Methods are now available to measure large-scale molecular, cellular, and circuit-level phenotypes, with additional methods in development.

    “Methods have recently become available that permit the measurement of large-scale molecular4,5, cellular6 and circuit-level3 phenotypes, and additional methods are currently in development7.”
  • Recent development of genetically encoded calcium indicators and innovations in imaging neuronal activity in awake, head-fixed mice have opened new opportunities toward studying structural-functional relationships.

    “The recent development of genetically encoded calcium indicators [89–91] and innovations in imaging neuronal activity in awake, head-fixed mice [92–96] have opened new opportunities toward this direction.”

    From Experience-dependent structural plasticity in the cortex (2011) · cites this paper as [89]

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Authors
Tian, L., Hires, S. A., Mao, T., Huber, D., Chiappe, M. E., Chalasani, S. H., Petreanu, L., Akerboom, J., McKinney, S. A., Schreiter, E. R., Bargmann, C. I., Jayaraman, V., Svoboda, K., & Looger, L. L.
Affiliations
  • Howard Hughes Medical Institute (US)
  • Stowers Institute for Medical Research (US)
  • University of Puerto Rico at Río Piedras (PR)
Venue
Nature Methods
Published
2009-11-08
Volume
6
Issue
12
Pages
875-881
ISSN
  • 1548-7091 (ISSN-L)
  • 1548-7105
OpenAlex
W2109234881
Cited by
1,967
References
46
Citations by year
2026: 46 · 2025: 68 · 2024: 84 · 2023: 93 · 2022: 100 · 2021: 98 · 2020: 122 · 2019: 111 · 2018: 134 · 2017: 127 · 2016: 132 · 2015: 152 · 2014: 179 · 2013: 197 · 2012: 172
FWCI
63.84
Type
journal article
Language
en
Open access
Locations
  • Nature Methods (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Europe PMC (PubMed Central) (acceptedVersion)
  • PubMed Central (acceptedVersion)
Funders
Howard Hughes Medical Institute
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
  • Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
  • Life Sciences › Neuroscience › Sensory Systems › Ion Channels and Receptors
Keywords
genetically encoded calcium indicators, GCaMP3, calcium imaging, neuronal activity imaging, dynamic range, photostability, calcium transients
MeSH terms
Animals, Brain, Calcium, Cell Line, Drosophila melanogaster, Humans, Neurons, Caenorhabditis elegans, Fluorescence Resonance Energy Transfer, Mice

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