Monitoring synaptic and neuronal activity in 3D with synthetic and genetic indicators using a compact acousto-optic lens two-photon microscope
Monitoring synaptic and neuronal activity in 3D with synthetic and genetic indicators using a compact acousto-optic lens two-photon microscope
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DOI：
10.1016/j.jneumeth.2013.10.021
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发表时间：
2014-01-30
影响因子：
3
通讯作者：
Silver, R. Angus
中科院分区：
文献类型：
--
作者：
Fernandez-Alfonso, Tomas;Nadella, K. M. Naga Srinivas;Silver, R. Angus
背景资料：双光子显微镜被广泛用于研究大脑功能，但传统的显微镜太慢，无法捕捉神经元信号的时间，成像仅限于一个平面。基于声光偏转器的随机访问功能成像的最新发展已经提高了时间分辨率，但是这些技术用于映射3D突触活动模式的效用及其在成像遗传编码指示物所需的激发波长处的性能尚未被研究。在这里，我们使用了一个紧凑的声光透镜（AOL）双光子显微镜，使高速[Ca 2 +]测量从棘和树突分布在三维与不同的激发波长结果：我们显示了使用合成的以及遗传编码的指示剂同时监测来自分布在神经元树突的3D树状结构上的许多突触输入的活动。我们确认的实用程序AOL为基础的成像快速在体内记录测量，同时，视觉诱发反应分布在150 pm的焦点深度范围内的100个神经元。此外，我们探索的方法来提高测量的神经元激活的时间通过选择特定区域内的细胞索马。与现有的方法比较：这些结果建立基于AOL的三维随机存取双光子显微镜具有更广泛的神经科学应用比以前所示。结论：我们的研究结果表明，紧凑的AOL显微镜设计具有速度，空间分辨率，灵敏度和波长灵活性，可以在单个试验中测量突触和神经元活动的3D模式。(C)2013作者Elsevier B.V.出版，保留所有权利。
Background: Two-photon microscopy is widely used to study brain function, but conventional microscopes are too slow to capture the timing of neuronal signalling and imaging is restricted to one plane. Recent development of acousto-optic-deflector-based random access functional imaging has improved the temporal resolution, but the utility of these technologies for mapping 3D synaptic activity patterns and their performance at the excitation wavelengths required to image genetically encoded indicators have not been investigated.New method: Here, we have used a compact acousto-optic lens (AOL) two-photon microscope to make high speed [Ca2+] measurements from spines and dendrites distributed in 3D with different excitation wavelengths (800-920nm).Results: We show simultaneous monitoring of activity from many synaptic inputs distributed over the 3D arborisation of a neuronal dendrite using both synthetic as well as genetically encoded indicators. We confirm the utility of AOL-based imaging for fast in vivo recordings by measuring, simultaneously, visually evoked responses in 100 neurons distributed over a 150 pm focal depth range. Moreover, we explore ways to improve the measurement of timing of neuronal activation by choosing specific regions within the cell soma.Comparison with existing methods: These results establish that AOL-based 3D random access two-photon microscopy has a wider range of neuroscience applications than previously shown. Conclusions: Our findings show that the compact AOL microscope design has the speed, spatial resolution, sensitivity and wavelength flexibility to measure 3D patterns of synaptic and neuronal activity on individual trials. (C) 2013 The Authors. Published by Elsevier B.V. All rights reserved.