Intact skull chronic windows for mesoscopic wide-field imaging in awake mice
Intact skull chronic windows for mesoscopic wide-field imaging in awake mice
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DOI：
10.1016/j.jneumeth.2016.04.012
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发表时间：
2016-07-15
影响因子：
3
通讯作者：
Murphy, Timothy H.
中科院分区：
文献类型：
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作者：
Silasi, Gergely;Xiao, Dongsheng;Murphy, Timothy H.
背景资料：基于颅骨切开术的窗口植入物通常用于头部固定啮齿动物的显微成像，但它们的视野通常很小，与皮层的介观功能映射不兼容。新方法：我们描述了一种可重复且简单的程序，用于清醒头部固定小鼠的慢性穿骨宽视野成像，为大面积皮层的慢性成像提供稳定的光学通路，持续数月。该制备是通过将清洁干燥的牙科粘固剂应用于完整的小鼠头骨，然后用玻璃盖玻片创建部分透明的成像表面来制备的。手术时间约30分钟。一个固定螺钉提供了一个稳定的连接方式（相对于测量的横向和轴向分辨率）进行中尺度评估，而不会模糊皮质视野。与现有方法的比较：我们证明了这种方法的实用性，通过显示种子-像素功能连接图产生于清醒和麻醉小鼠中GCAMP 6信号的自发皮质活动，纵向研究长达2个月的持续时间。结论：我们建议，这里描述的完整的颅骨制备可用于大多数纵向研究，不需要微米级分辨率和皮质神经或血管信号记录与内在传感器或在转基因小鼠表达基因编码的传感器的活动。(C)2016爱思唯尔B. V.保留所有权利。
Background: Craniotomy-based window implants are commonly used for microscopic imaging, in head fixed rodents, however their field of view is typically small and incompatible with mesoscopic functional mapping of cortex.New method: We describe a reproducible and simple procedure for chronic through-bone wide-field imaging in awake head-fixed mice providing stable optical access for chronic imaging over large areas of the cortex for months.Results: The preparation is produced by applying clear-drying dental cement to the intact mouse skull, followed by a glass coverslip to create a partially transparent imaging surface. Surgery time takes about 30 min. A single set-screw provides a stable means of attachment (in relation to the measured lateral and axial resolution) for mesoscale assessment without obscuring the cortical field of view.Comparison with existing methods: We demonstrate the utility of this method by showing seed-pixel functional connectivity maps generated from spontaneous cortical activity of GCAMP6 signals in both awake and anesthetized mice in longitudinal studies of up to 2 months in duration.Conclusions: We propose that the intact skull preparation described here may be used for most longitudinal studies that do not require micron scale resolution and where cortical neural or vascular signals are recorded with intrinsic sensors or in transgenic mice expressing genetically encoded sensors of activity. (C) 2016 Elsevier B.V. All rights reserved.