| First Author | Akerboom J | Year | 2012 |
| Journal | J Neurosci | Volume | 32 |
| Issue | 40 | Pages | 13819-40 |
| PubMed ID | 23035093 | Mgi Jnum | J:223216 |
| Mgi Id | MGI:5648499 | Doi | 10.1523/JNEUROSCI.2601-12.2012 |
| Citation | Akerboom J, et al. (2012) Optimization of a GCaMP calcium indicator for neural activity imaging. J Neurosci 32(40):13819-40 |
| abstractText | 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. |
