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dc.contributor.authorAkerboom, Jasper
dc.contributor.authorChen, Tsai-Wen
dc.contributor.authorWardill, Trevor J.
dc.contributor.authorTian, Lin
dc.contributor.authorMarvin, Jonathan S.
dc.contributor.authorMutlu, Sevinc
dc.contributor.authorCalderon, Nicole Carreras
dc.contributor.authorEsposti, Federico
dc.contributor.authorBorghuis, Bart G.
dc.contributor.authorSun, Xiaonan Richard
dc.contributor.authorGordus, Andrew
dc.contributor.authorOrger, Michael B.
dc.contributor.authorPortugues, Ruben
dc.contributor.authorEngert, Florian
dc.contributor.authorMacklin, John J.
dc.contributor.authorFilosa, Alessandro
dc.contributor.authorAggarwal, Aman
dc.contributor.authorKerr, Rex A.
dc.contributor.authorTakagi, Ryousuke
dc.contributor.authorKracun, Sebastian
dc.contributor.authorShigetomi, Eiji
dc.contributor.authorKhakh, Baljit S.
dc.contributor.authorBaier, Herwig
dc.contributor.authorLagnado, Leon
dc.contributor.authorWang, Samuel S.-H.
dc.contributor.authorBargmann, Cornelia I.
dc.contributor.authorKimmel, Bruce E.
dc.contributor.authorJayaraman, Vivek
dc.contributor.authorSvoboda, Karel
dc.contributor.authorKim, Douglas S.
dc.contributor.authorSchreiter, Eric R.
dc.contributor.authorLooger, Loren L.
dc.identifier.citationJournal of Neuroscience 32 (2012): 13819-13840en_US
dc.description© The Author(s), 2012. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Journal of Neuroscience 32 (2012): 13819-13840, doi:10.1523/JNEUROSCI.2601-12.2012.en_US
dc.description.abstractGenetically 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 in vivo. Through protein structure determination, targeted mutagenesis, high-throughput screening, and a battery of in vitro 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 in vivo in Caenorhabditis chemosensory neurons, Drosophila 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 in vivo 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 in vivo and may find widespread applications for cellular imaging in general.en_US
dc.description.sponsorshipA.F. has been supported by a European Molecular Biology Organization long-term fellowship. Work in H.B.’s laboratory was funded by the National Institutes of Health (NIH) Nanomedicine Development Center “Optical Control of Biological Function,” and work in S.S.-H.W.’s laboratory was funded by NIH R01 NS045193.en_US
dc.publisherSociety for Neuroscienceen_US
dc.titleOptimization of a GCaMP calcium indicator for neural activity imagingen_US

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