Kim Douglas S.

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Douglas S.

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  • Article
    Optimization of a GCaMP calcium indicator for neural activity imaging
    (Society for Neuroscience, 2012-10-03) Akerboom, Jasper ; Chen, Tsai-Wen ; Wardill, Trevor J. ; Tian, Lin ; Marvin, Jonathan S. ; Mutlu, Sevinc ; Calderon, 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 ; Wang, Samuel S.-H. ; Bargmann, Cornelia I. ; Kimmel, Bruce E. ; Jayaraman, Vivek ; Svoboda, Karel ; Kim, Douglas S. ; Schreiter, Eric R. ; Looger, Loren L.
    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 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.