Department of Zoology and Physiology, University of Wyoming.
School of Electrical Engineering & Computer Science, University of North Dakota.
J Vis Exp. 2021 Oct 8(176). doi: 10.3791/63049.
A miniature fluorescence microscope (miniscope) is a potent tool for in vivo calcium imaging from freely behaving animals. It offers several advantages over conventional multi-photon calcium imaging systems: (1) compact; (2) light-weighted; (3) affordable; and (4) allows recording from freely behaving animals. This protocol describes brain surgeries for deep brain in vivo calcium imaging using a custom-developed miniscope recording system. The preparation procedure consists of three steps, including (1) stereotaxically injecting the virus at the desired brain region of a mouse brain to label a specific subgroup of neurons with genetically encoded calcium sensor; (2) implantation of gradient-index (GRIN) lens that can relay calcium image from deep brain region to the miniscope system; and (3) affixing the miniscope holder over the mouse skull where miniscope can be attached later. To perform in vivo calcium imaging, the miniscope is fastened onto the holder, and neuronal calcium images are collected along with simultaneous behavior recordings. The present surgery protocol is compatible with any commercial or custom-built single-photon and two-photon imaging systems for deep brain in vivo calcium imaging.
微型荧光显微镜（miniscope）是一种强大的工具，可用于对自由活动的动物进行体内钙成像。它与传统的多光子钙成像系统相比具有几个优势：（1）紧凑；（2）重量轻；（3）价格合理；（4）允许对自由活动的动物进行记录。本协议描述了使用定制的 miniscope 记录系统进行深部脑内体内钙成像的脑部手术。准备程序包括三个步骤，包括（1）在小鼠大脑的预期脑区立体定向注射病毒，以用遗传编码的钙传感器标记特定的神经元亚群；（2）植入梯度折射率（GRIN）透镜，可将深部脑区的钙图像传递到 miniscope 系统；（3）将 miniscope 固定器固定在小鼠颅骨上，以后可以将 miniscope 固定在其上。要进行体内钙成像，将 miniscope 固定在固定器上，并同时进行神经元钙成像和行为记录。本手术方案与任何用于深部脑内体内钙成像的商业或定制的单光子和双光子成像系统兼容。
