RT Journal Article
SR Electronic
T1 Optimization of a GCaMP Calcium Indicator for Neural Activity Imaging
JF The Journal of Neuroscience
JO J. Neurosci.
FD Society for Neuroscience
SP 13819
OP 13840
DO 10.1523/JNEUROSCI.2601-12.2012
VO 32
IS 40
A1 Jasper Akerboom
A1 Tsai-Wen Chen
A1 Trevor J. Wardill
A1 Lin Tian
A1 Jonathan S. Marvin
A1 Sevinç Mutlu
A1 Nicole Carreras Calderón
A1 Federico Esposti
A1 Bart G. Borghuis
A1 Xiaonan Richard Sun
A1 Andrew Gordus
A1 Michael B. Orger
A1 Ruben Portugues
A1 Florian Engert
A1 John J. Macklin
A1 Alessandro Filosa
A1 Aman Aggarwal
A1 Rex A. Kerr
A1 Ryousuke Takagi
A1 Sebastian Kracun
A1 Eiji Shigetomi
A1 Baljit S. Khakh
A1 Herwig Baier
A1 Leon Lagnado
A1 Samuel S.-H. Wang
A1 Cornelia I. Bargmann
A1 Bruce E. Kimmel
A1 Vivek Jayaraman
A1 Karel Svoboda
A1 Douglas S. Kim
A1 Eric R. Schreiter
A1 Loren L. Looger
YR 2012
UL http://www.jneurosci.org/content/32/40/13819.abstract
AB 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.