Fast two-layer two-photon imaging of neuronal cell populations using an electrically tunable lens.
Fast two-layer two-photon imaging of neuronal cell populations using an electrically tunable lens.
复制标题
DOI：
10.1364/boe.2.002035
复制
发表时间：
2011-07-01
影响因子：
3.4
通讯作者：
Helmchen F
中科院分区：
文献类型：
--
作者：
Grewe BF;Voigt FF;van 't Hoff M;Helmchen F
功能双光子Ca2+成像是研究脑切片和活体动物神经元种群动态的多功能工具。然而，群体成像通常局限于单个二维成像平面。通过在双光子显微镜的激发路径中引入电可调透镜，我们能够在15 ms内实现快速轴向焦点移动。最大轴向扫描范围为0.7毫米，采用40倍NA0.8水浸泡物镜，通常需要的范围为0.2-0.3毫米。通过将轴向扫描方法与二维声光框架扫描和随机存取扫描相结合，我们在间隔40 μm的两个成像平面上测量了约40个神经元的神经元群活动，并实现了高达20-30 Hz的扫描速率。所提出的方法易于应用，并允许升级现有的双光子显微镜进行快速三维扫描。
Functional two-photon Ca2+-imaging is a versatile tool to study the dynamics of neuronal populations in brain slices and living animals. However, population imaging is typically restricted to a single two-dimensional image plane. By introducing an electrically tunable lens into the excitation path of a two-photon microscope we were able to realize fast axial focus shifts within 15 ms. The maximum axial scan range was 0.7 mm employing a 40x NA0.8 water immersion objective, plenty for typically required ranges of 0.2–0.3 mm. By combining the axial scanning method with 2D acousto-optic frame scanning and random-access scanning, we measured neuronal population activity of about 40 neurons across two imaging planes separated by 40 μm and achieved scan rates up to 20–30 Hz. The method presented is easily applicable and allows upgrading of existing two-photon microscopes for fast 3D scanning.