| First Author | Forli A | Year | 2021 |
| Journal | Elife | Volume | 10 |
| PubMed ID | 34032211 | Mgi Jnum | J:327580 |
| Mgi Id | MGI:6728251 | Doi | 10.7554/eLife.63359 |
| Citation | Forli A, et al. (2021) Optogenetic strategies for high-efficiency all-optical interrogation using blue-light-sensitive opsins. Elife 10:e63359 |
| abstractText | All-optical methods for imaging and manipulating brain networks with high spatial resolution are fundamental to study how neuronal ensembles drive behavior. Stimulation of neuronal ensembles using two-photon holographic techniques requires high-sensitivity actuators to avoid photodamage and heating. Moreover, two-photon-excitable opsins should be insensitive to light at wavelengths used for imaging. To achieve this goal, we developed a novel soma-targeted variant of the large-conductance blue-light-sensitive opsin CoChR (stCoChR). In the mouse cortex in vivo, we combined holographic two-photon stimulation of stCoChR with an amplified laser tuned at the opsin absorption peak and two-photon imaging of the red-shifted indicator jRCaMP1a. Compared to previously characterized blue-light-sensitive soma-targeted opsins in vivo, stCoChR allowed neuronal stimulation with more than 10-fold lower average power and no spectral crosstalk. The combination of stCoChR, tuned amplified laser stimulation, and red-shifted functional indicators promises to be a powerful tool for large-scale interrogation of neural networks in the intact brain. |
