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

在这笔赠款的第一年，在建立必要的神经科学设置方面取得了实质性进展，例如结合空间光调制的双光子成像(2pi)。 Plenz实验室在技术开发方面取得了以下进展： 在双光子成像方面，我们实现了最新一代遗传编码钙指示剂(GECI)的稳健表达和使用，特别是jGCaMP7和YC2.0。我们还升级了目前的成像激光器，以包括波长&1000 nm，以供将来使用红移的GECI。此外，我们现在能够将我们的2pi系统与光刺激、斜度测量和移动措施同步，用于常规记录。 在我们的空间光调制(SLM)/光刺激方面，我们优化了红移Opsins(C1V1，Chrimson)的注射和表达程序，并通过同时进行2pi的大范围Opsins刺激在清醒小鼠体内展示了功能性视蛋白的表达(摘要提交给SFN 2019)。此外，我们实施了定制的SLM路径，并将其与我们现有的2pi设置对准，用于体内光学刺激。在软件控制方面，我们利用现有的胸腔成像软件建立了SLM的软件控制，用于刺激定位和持续时间控制。我们还建立了体外急性切片记录装置来改进视蛋白筛查，并实施了硬件/程序来功能性地识别觉醒小鼠的初级视觉皮质。

During the first year of this grant substantial progress has been made in setting up 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: With respect to 2-photon imaging, we implemented a robust expression and use of last generation genetically encoded calcium indicators (GECIs), specifically jGCaMP7s and YC2.0. We also upgraded our current imaging laser to include wavelengths >1000 nm for future use of red-shifted GECIs. 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 (C1V1, Chrimson) and demonstrated functional opsin expression in vivo in the awake mouse using wide-field stimulation of opsins with simultaneous 2PI (Abstract submitted to SfN 2019). Furthermore, we implemented and aligned a custom SLM path with our existing 2PI set-up for in vivo optical stimulation. Regarding software control, we established software control of SLM with our existing Thorlabs imaging software for stimulus location and duration control. We also established in vitro acute slice recording set-up to improve opsin screening and implemented hardware/procedures to functionally identify primary visual cortex in wake mice.