Szalay Gergely, Judák Linda, Katona Gergely, Ócsai Katalin, Juhász Gábor, Veress Máté, Szadai Zoltán, Fehér András, Tompa Tamás, Chiovini Balázs, Maák Pál, Rózsa Balázs
Laboratory of 3D Functional Network and Dendritic Imaging, Institute of Experimental Medicine, Hungarian Academy of Sciences, Budapest 1083, Hungary.
Laboratory of 3D Functional Network and Dendritic Imaging, Institute of Experimental Medicine, Hungarian Academy of Sciences, Budapest 1083, Hungary; MTA-PPKE ITK-NAP B-2p Measurement Technology Group, Faculty of Information Technology, Pázmány Péter Catholic University, Budapest 1083, Hungary.
Neuron. 2016 Nov 23;92(4):723-738. doi: 10.1016/j.neuron.2016.10.002. Epub 2016 Oct 20.
Understanding neural computation requires methods such as 3D acousto-optical (AO) scanning that can simultaneously read out neural activity on both the somatic and dendritic scales. AO point scanning can increase measurement speed and signal-to-noise ratio (SNR) by several orders of magnitude, but high optical resolution requires long point-to-point switching time, which limits imaging capability. Here we present a novel technology, 3D DRIFT AO scanning, which can extend each scanning point to small 3D lines, surfaces, or volume elements for flexible and fast imaging of complex structures simultaneously in multiple locations. Our method was demonstrated by fast 3D recording of over 150 dendritic spines with 3D lines, over 100 somata with squares and cubes, or multiple spiny dendritic segments with surface and volume elements, including in behaving animals. Finally, a 4-fold improvement in total excitation efficiency resulted in about 500 × 500 × 650 μm scanning volume with genetically encoded calcium indicators (GECIs).
要理解神经计算，需要诸如三维声光（AO）扫描之类的方法，这些方法能够同时在体细胞和树突尺度上读出神经活动。AO点扫描可将测量速度和信噪比（SNR）提高几个数量级，但高光学分辨率需要较长的点对点切换时间，这限制了成像能力。在此，我们展示了一种新技术——三维漂移AO扫描，它可以将每个扫描点扩展为小的三维线、面或体素，以便在多个位置同时对复杂结构进行灵活快速的成像。我们的方法通过以下方式得到了验证：用三维线对150多个树突棘进行快速三维记录，用正方形和立方体对100多个体细胞进行记录，或用表面和体素对多个多刺树突段进行记录，包括在行为动物中。最后，总激发效率提高了4倍，使用基因编码钙指示剂（GECIs）可实现约500×500×650μm的扫描体积。
