BRAIN project (Plenz): Readout and Control of Spatiotemporal Neuronal Codes of Behavior

在该资助的第二年，我们继续在优化我们所需的神经科学设置方面取得实质性进展，例如结合空间光调制的双光子成像（2 PI）。 Plenz实验室在技术开发方面取得的进展如下： 在将我们目前的成像激光器扩展到包括波长>1000 nm之后，我们用红移GECIs（如jrGecko）建立了深层皮层记录。此外，我们现在能够将我们的2 PI系统与光学刺激、瞳孔测量和运动测量同步，以进行常规记录。 关于我们的空间光调制（SLM）/光学刺激，我们优化了红移视蛋白（Chrimson）的注射和表达程序，并使用同时2 PI的视蛋白的宽场刺激在清醒小鼠中展示了体内功能性视蛋白表达。我们进一步优化了SLM的软件控制，并将SLM升级为能够以更高的空间分辨率控制更高功率激光的型号。 最后，我们用低重复、高功率的双激光器取代了高重复的5 W刺激，以便能够以毫秒级的精度刺激单个神经细胞，这对于控制清醒动物大脑中的关键动力学非常重要。

During the second year of this grant we continued to make substantial progress in optimizing our required neuroscientific set-ups such as 2-photon imaging (2PI) in combination with spatial light modulation. Advances in technical development specific to the Plenz lab were the following: After upgradeding our current imaging laser to include wavelengths >1000 nm, we established deep cortical recording with red-shifted GECIs such as jrGecko. Furthermore, we are now able to synchronize our 2PI system with optical stimulation, pupillometry and locomotion measures for routine recordings. With respect to our Spatial Light Modulation (SLM)/optical stimulation, we optimized injection and expression procedures for red-shifted opsins (Chrimson) and demonstrated functional opsin expression in vivo in the awake mouse using wide-field stimulation of opsins with simultaneous 2PI. We further optimized our software control of the SLM and upgraded our SLM to a model that can control lasers with higher power at higher spatial resolution. Finally, we replaced our high-repetition 5 W stimulation with a low-repetition, high power dual laser to be able to stimulation individual nerve cells with millisecond precision important to control critical dynamics in the brain of awake animals.