Quasi-simultaneous multiplane calcium imaging of neuronal circuits
Quasi-simultaneous multiplane calcium imaging of neuronal circuits
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DOI：
10.1364/boe.10.000267
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发表时间：
2019-01-01
影响因子：
3.4
通讯作者：
Booth, Martin J.
中科院分区：
文献类型：
--
作者：
Chong, Ee Zhuan;Panniello, Mariangela;Booth, Martin J.
双光子激发荧光显微镜被广泛用于研究神经元回路的活动。然而，对于标准显微镜设计，快速成像通常被约束到单个横向平面。鉴于小鼠大脑中的皮质神经元网络是组织在六个组织学定义的层中的复杂三维结构，这些层延伸超过数百微米，因此对能够大量记录神经元信号的显微镜系统有强烈的需求。阻碍，我们开发了一种准同时多平面成像技术结合声光偏转器和静态远程聚焦，以提供快速成像的神经元从不同的轴向位置内的皮质层，而不需要机械干扰的目标透镜或标本。硬件和软件很容易适应现有的双光子显微镜。在这里，我们证明了我们的成像方法可以记录，在高速和高图像对比度，在两个不同的成像平面轴向分离的焦点和重聚焦平面分别低于大脑表面120 μ m和250 μ m的神经元的钙动力学。由The Optical Society根据Creative Commons Attribution 4.0 License条款发布。
Two-photon excitation fluorescence microscopy is widely used to study the activity of neuronal circuits. However, the fast imaging is typically constrained to a single lateral plane for a standard microscope design. Given that cortical neuronal networks in a mouse brain are complex three-dimensional structures organised in six histologically defined layers which extend over many hundreds of micrometres, there is a strong demand for microscope systems that can record neuronal signalling in volumes. Henceforth, we developed a quasi-simultaneous multiplane imaging technique combining an acousto-optic deflector and static remote focusing to provide fast imaging of neurons from different axial positions inside the cortical layers without the need for mechanical distuibance of either the objective lens or the specimen. The hardware and the software are easily adaptable to existing two-photon microscopes. Here, we demonstrated that our imaging method can record, at high speed and high image contrast, the calcium dynamics of neurons in two different imaging planes separated axially with the in-focus and the refocused planes 120 mu m and 250 mu m below the brain surface respectively. Published by The Optical Society under the terms of the Creative Commons Attribution 4.0 License.