Fu Ting, Arnoux Isabelle, Döring Jan, Backhaus Hendrik, Watari Hirofumi, Stasevicius Ignas, Fan Wei, Stroh Albrecht
Institute of Pathophysiology, University Medical Center Mainz, Hanns-Dieter-Hüsch-Weg 19, D-55128 Mainz, Germany.
Leibniz Institute for Resilience Research, Wallstr. 7, D-55122 Mainz, Germany.
iScience. 2021 Feb 12;24(3):102184. doi: 10.1016/j.isci.2021.102184. eCollection 2021 Mar 19.
Two-photon (2-P) all-optical approaches combine 2-P calcium imaging and 2-P optogenetic modulations. Here, firstly, we combined juxtacellular recordings and GCaMP6f-based 2-P calcium imaging in mouse visual cortex to tune our detection algorithm towards a 100% specific identification of action potential-related calcium transients. Secondly, we minimized photostimulation artifacts by using extended-wavelength-spectrum laser sources for optogenetic stimulation. We achieved artifact-free all-optical experiments performing optogenetic stimulation from 1100 nm to 1300 nm. Thirdly, we determined the spectral range for maximizing efficacy until 1300 nm. The rate of evoked transients in GCaMP6f/C1V1-co-expressing cortical neurons peaked already at 1100 nm. By refining spike detection and defining 1100 nm as the optimal wavelength for artifact-free and effective GCaMP6f/C1V1-based all-optical physiology, we increased the translational value of these approaches, e.g., for the development of network-based therapies.
双光子（2-P）全光学方法结合了双光子钙成像和双光子光遗传学调制。在此，首先，我们将小鼠视觉皮层中的细胞旁记录与基于GCaMP6f的双光子钙成像相结合，调整我们的检测算法，以实现对动作电位相关钙瞬变的100%特异性识别。其次，我们通过使用用于光遗传学刺激的扩展波长光谱激光源来最小化光刺激伪影。我们通过在1100纳米至1300纳米范围内进行光遗传学刺激，实现了无伪影的全光学实验。第三，我们确定了在1300纳米之前使功效最大化的光谱范围。在共表达GCaMP6f/C1V1的皮层神经元中，诱发瞬变率在1100纳米时已达到峰值。通过改进尖峰检测并将1100纳米定义为基于GCaMP6f/C1V1的无伪影且有效的全光学生理学的最佳波长，我们提高了这些方法的转化价值，例如在基于网络的疗法开发方面。
