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Akerboom, Jasper; Chen, Tsai-Wen; Wardill, Trevor J.; Tian, Lin; Marvin, Jonathan S.; Mutlu, Sevinç; Calderón, Nicole Carreras; Esposti, Federico; Borghuis, Bart G.; Sun, Xiaonan Richard; Gordus, Andrew; Orger, Michael B.; Portugues, Ruben; Engert, Florian; Macklin, John J.; Filosa, Alessandro; Aggarwal, Aman; Kerr, Rex A.; Takagi, Ryousuke; Kracun, Sebastian; Shigetomi, Eiji; Khakh, Baljit S.; Baier, Herwig; Lagnado, Leon; [...]

Optimization of a GCaMP Calcium Indicator for Neural Activity Imaging

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  • Media type: E-Article
  • Title: Optimization of a GCaMP Calcium Indicator for Neural Activity Imaging
  • Contributor: Akerboom, Jasper; Chen, Tsai-Wen; Wardill, Trevor J.; Tian, Lin; Marvin, Jonathan S.; Mutlu, Sevinç; Calderón, Nicole Carreras; Esposti, Federico; Borghuis, Bart G.; Sun, Xiaonan Richard; Gordus, Andrew; Orger, Michael B.; Portugues, Ruben; Engert, Florian; Macklin, John J.; Filosa, Alessandro; Aggarwal, Aman; Kerr, Rex A.; Takagi, Ryousuke; Kracun, Sebastian; Shigetomi, Eiji; Khakh, Baljit S.; Baier, Herwig; Lagnado, Leon; [...]
  • imprint: Society for Neuroscience, 2012
  • Published in: The Journal of Neuroscience
  • Language: English
  • DOI: 10.1523/jneurosci.2601-12.2012
  • ISSN: 0270-6474; 1529-2401
  • Origination:
  • Footnote:
  • Description: <jats:p>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<jats:italic>in vivo</jats:italic>. Through protein structure determination, targeted mutagenesis, high-throughput screening, and a battery of<jats:italic>in vitro</jats:italic>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<jats:italic>in vivo</jats:italic>in<jats:italic>Caenorhabditis</jats:italic>chemosensory neurons,<jats:italic>Drosophila</jats:italic>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<jats:italic>in vivo</jats:italic>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<jats:italic>in vivo</jats:italic>and may find widespread applications for cellular imaging in general.</jats:p>
  • Access State: Open Access