Name: two-photon calcium imaging in mice navigating a virtual reality environment
Uploaded: 2014-02-20T22:00:00
Description: Watch this Scientific Journal Video about Two-photon Calcium Imaging in Mice Navigating a Virtual Reality Environment at JoVE.com

这项工作是由弗里德里希·米歇尔生物医学研究所，马普学会，和人类前沿科学计划的支持。

所有动物的程序获得批准，并按照广巴塞尔城市州的兽医部门的指导下进行。 1。硬件和软件设置<ol><li>双光子显微镜的设置： <ol><li>用脉冲红外激光作为照明源（脉冲宽度&lt;120飞秒）。 </li><li>使用一个扫描头8或12 kHz的谐振扫描器和一个标准的振镜注组成：这使40或60赫兹的帧频在750×400像素。高帧速率对于减少脑运动诱发的图像失真的关键。此外?...

<ol>
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的关键行为的双光子成像的成功是制备在两个方面的稳定性： <ol><li>以上的天交窗口植入的过程中，组织的炎症反应可以导致增强肉芽组织和软骨会阻碍或什至阻止成像的形成。 </li><li>在实验过程中的大脑有足够稳定，以防止运动伪影败坏神经活动相关的荧光信号。 </li></ol>为了保持到最低限度颅窗口植入术中应尽可能快地和直接向脑损伤进行了炎症免疫反应，应不惜一切代...

钙指标（基因编码的像GCaMP5 7或R-GECO 8，或合成染料像OGB或Fluo4）双光子成像已经成为测量神经元活动的行为小鼠1-6的有力方法。它使数百个细胞的在接近单个动作电位分辨率活动的同时进行测量，最多约800微米的脑表面9,10所示。此外，使用基因编码的钙指标（GECIS）神经元的活性可测慢性5,11,12，和在遗传背景明确细胞类型13。总之，这些方法提供了一定程度的打开了大量的神经元计算的体内研究中新的可能性，时间和空间分辨率。 手术治疗是必要的揭露和标记的小鼠大脑进行成像。细胞使用的是重组腺相关vir区典型地转<html>我们（AAV）系统GECI送货和颅骨窗被植入在注射部位获得的光纤接入到大脑。甲头杆然后被连接到双光子显微镜下颅骨为头部固定。因为大多数的问题，清醒成像产生于准备不稳定，这些步骤的设计和实施是至关重要的。理想的情况是在这里描述的过程应该允许多达几个月后手术慢性成像。 为了在双光子成像使视觉引导的行为，头部固定鼠标坐在一个支持空气球跑步机，它可以用它来浏览虚拟现实环境。鼠标在跑步机上的运动被连接到运动中所显示周围鼠标14,15的环形屏幕上的虚拟环境。行为变量如运动，视觉刺激，而瞳孔的位置可以记录6。 T“&gt;我们描述参与了小鼠探索虚拟现实环境慢性双光子成像的程序解决的关键点是：减少运动伪影，减少漏光，同时记录细胞数量的最大化，并最小化光损伤，我们还提供有关设置空中支持的跑步机，瞳孔跟踪，以及虚拟现实环境的细节。这里所描述的程序，可以用于成像的头部固定的小鼠荧光标记的细胞群中潜在的各种行为范式。

<table border="0" cellpadding="0" cellspacing="0" width="653">
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		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="20">
			<td height="20" style="height:20px;width:216px;">cover slips (d = 3-5 mm)</td>
			<td style="width:151px;">Menzel</td>
			<td style="width:119px;"></td>
			<td style="width:167px;">window implant</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">InSight DeepSee laser</td>
			<td>Spectra-Physics</td>
			<td></td>
			<td>microscope</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">12kHz resonance scanner</td>
			<td>Cambridge Technology</td>
			<td>G1-003-30026</td>
			<td>microscope</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Galvometer</td>
			<td>Cambridge Technology</td>
			<td>G6215H</td>
			<td>microscope</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Digitizer</td>
			<td>National Instruments</td>
			<td>NI 5772</td>
			<td>microscope</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">FPGA</td>
			<td>National Instruments</td>
			<td>PXIe 7965R</td>
			<td>microscope</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Acquisition card</td>
			<td>National Instruments</td>
			<td>PCIe 6363</td>
			<td>microscope</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Emission filter 525/50</td>
			<td>Semrock</td>
			<td>FF03-525/50-25</td>
			<td>microscope</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Piezo-electric z-drive</td>
			<td>Physikinstrumente</td>
			<td>P-726.1CD</td>
			<td>microscope</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Controller for piezo-electric drive</td>
			<td>Physikinstrumente</td>
			<td>E665 LVPZT</td>
			<td>microscope</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Objective 16x, 0.8NA</td>
			<td>Nikon</td>
			<td>CFI75</td>
			<td>microscope</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:216px;">Current amplifier</td>
			<td style="width:151px;">Femto</td>
			<td style="width:119px;">DHPCA-100</td>
			<td>microscope</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:216px;">Photomultiplier tube</td>
			<td style="width:151px;">Hamamatsu</td>
			<td style="width:119px;"></td>
			<td>microscope</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">USB Camera without IR filter</td>
			<td>ImagingSource</td>
			<td>DMK22BUC03&#160;</td>
			<td>pupil tracking</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Objective 50 mm</td>
			<td>ImagingSource</td>
			<td>M5018-MP&#160;</td>
			<td>pupil tracking</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Macro adapter rings</td>
			<td>ImagingSource</td>
			<td>LAexSet</td>
			<td>pupil tracking</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Optical computer mouse</td>
			<td>Logitech</td>
			<td>G500</td>
			<td>motion tracking</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Styrofoam ball 20 cm</td>
			<td>e.g. idee-shop.de</td>
			<td>08797.00.15</td>
			<td>virtual environment</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">LED projector</td>
			<td>Samsung</td>
			<td>SP-F10M&#160;</td>
			<td>virtual environment</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Acquisition card</td>
			<td>National Instruments</td>
			<td style="width:119px;">NI 6009</td>
			<td>virtual environment</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Panda3D game engine</td>
			<td></td>
			<td><a href="http://www.panda3d.org/">www.panda3d.org</a></td>
			<td>virtual environment</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:216px;">Numpy library for Python</td>
			<td style="width:151px;"></td>
			<td style="width:119px;"><a href="http://www.scipy.org/">www.scipy.org</a></td>
			<td>virtual environment</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:216px;">Scipy library for Python</td>
			<td style="width:151px;"></td>
			<td style="width:119px;"><a href="http://www.scipy.org/">www.scipy.org</a></td>
			<td>virtual environment</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:216px;">NI-DAQmx driver</td>
			<td style="width:151px;">National Instruments</td>
			<td style="width:119px;"><a href="http://www.ni.com/">www.ni.com</a></td>
			<td>virtual environment</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:216px;">Ultrasound gel</td>
			<td style="width:151px;">Dahlhausen</td>
			<td style="width:119px;">5701.0342.10</td>
			<td>imaging</td>
		</tr>
	</tbody>
</table>

two-photon calcium imaging in mice navigating a virtual reality environment

近年来，双光子成像已经成为神经科学中一个宝贵的工具，因为它允许基因鉴定细胞的行为1-6中的活性慢性测量。这里我们描述了执行双光子成像技术在小鼠皮质，而动物导航虚拟现实环境的方法。我们注重的实验程序，是关键的行为动物成像在明亮的虚拟环境的各个方面。过程中出现的这个实验装置，我们这里的地址是关键的问题：尽量减少大脑运动有关的文物，最大限度地减少从虚拟现实投影系统漏光，并尽量减少激光诱导组织损伤。我们也提供来样软件来控制虚拟现实环境，并做瞳孔跟踪。这些程序和资源应该是可能要转换的常规双光子显微镜使用中表现小鼠。

在标有GECI细胞群的双光子钙成像的图像质量在很大程度上依赖于颅窗口植入物的质量。二周后病毒注射颅窗口应检查清楚。不应该有肉芽组织或骨再生长可见（ 图1A）。此外，浅表血管的图案应该保持不变和脉管系统的边界应当清晰明确。在同一时间，GECI表达也可以进行检查。标记细胞勃利应该是清晰可见的落射荧光显微镜（ 图1B）下。 理想的情?...

Watch this Scientific Journal Video about Two-photon Calcium Imaging in Mice Navigating a Virtual Reality Environment at JoVE.com

Two-photon Calcium Imaging in Mice Navigating a Virtual Reality Environment

这里，我们描述在一个虚拟现实环境中的行为过程中涉及的小鼠皮质的双光子成像的实验程序。

在小鼠导航虚拟现实环境的双光子钙成像

In recent years, two-photon imaging has become an invaluable tool in neuroscience, as it allows for chronic measurement of the activity of genetically ...

Research

JoVE Journal

Behavior

Development of a Virtual Reality Assessment of Everyday Living Skills

每天生活技能的虚拟现实评估的发展

Cognitive impairments affect the majority of patients with schizophrenia and these impairments predict poor long term psychosocial outcomes.&#160; ...

科研

行为学

The Modified Hole Board - Measuring Behavior, Cognition and Social Interaction in Mice and Rats

修改后的孔板 - 测量行为，认知和社会互动小鼠和大鼠

This protocol describes the modified hole board (mHB), which combines features from a traditional hole board and open field and is designed to measure ...

A Protocol for Housing Mice in an Enriched Environment

在丰富环境的协议房屋小鼠

Environmental enrichment (EE) is a housing environment for mice that boosts mental and physical health compared to standard laboratory housing. Our recent ...

Multifunctional Setup for Studying Human Motor Control Using Transcranial Magnetic Stimulation, Electromyography, Motion Capture, and Virtual Reality

多功能设置研究人类电机控制用经颅磁刺激，肌电图，运动捕捉和虚拟现实

The study of neuromuscular control of movement in humans is accomplished with numerous technologies. Non-invasive methods for investigating neuromuscular ...

Using Virtual Reality to Transfer Motor Skill Knowledge from One Hand to Another

利用虚拟现实技术将运动技能知识从一只手转移到另一个

As far as acquiring motor skills is concerned, training by voluntary physical movement is superior to all other forms of training (e.g. training by ...

Mobile Game-based Virtual Reality Program for Upper Extremity Stroke Rehabilitation

基于移动游戏的上肢卒中康复虚拟现实程序

Stroke rehabilitation requires repetitive, intensive, goal-oriented therapy. Virtual reality (VR) has the potential to satisfy these requirements. ...

A Method to Study Adaptation to Left-Right Reversed Audition

一种研究左右反试镜的适应方法

An unusual sensory space is one of the effective tools to uncover the mechanism of adaptability of humans to a novel environment. Although most of the ...

Measuring the Kinematics of Daily Living Movements with Motion Capture Systems in Virtual Reality

虚拟现实中运动捕捉系统的日常生活运动运动学测量

The inability to complete instrumental activities of daily living (IADL) is a precursor to various neuropsychological diseases. Questionnaire-based ...

A Networked Desktop Virtual Reality Setup for Decision Science and Navigation Experiments with Multiple Participants

面向多参与者的决策科学与导航实验的网络化桌面虚拟现实设置

Investigating the interactions among multiple participants is a challenge for researchers from various disciplines, including the decision sciences and ...