作者希望感谢捐赠者及其家人慷慨的馈赠。感谢您对 Mr. 小玄李录制视频并帮助进行视频编辑;Ms. 汉娜刘易斯和 Mr. 路易斯萨博提供技术支持;1月, m 教授对视频内容进行了审查并提供了输入。

此处描述的所有方法均已获得澳大利亚国立大学人类研究伦理委员会的批准。创建心室模型我们使用了科林格勒尔纤维解剖技术 12 , 14 。科林格勒尔技术是一种触觉解剖方法, 包括去除皮层中的小部分灰质和剥离神经纤维束, 从而提供从表面到深层结构的组织层的 step-by 步骤指南。的大脑. 
 注意: 在随附的视频和图像中, 用于演示此协议的大脑标本被小心地从来自医学学校的身体捐献者的尸体上取出, 从澳大利亚国立大学。捐赠者没有已知的病理疾病史。取出硬脑膜后, 大脑在室温下储存10% 乙醇溶液三年. 
 1. 准备 <ol> <li> 从被防腐的人的尸体上获取一个完整的大脑, 取出硬脑膜, 然后在解剖前在室温下用10% 乙醇储存大脑. 
	警告: 在通风良好的室内使用个人防护设备, 在处理时应遵照当地的指导方针。在开始解剖协议之前, 确保所有参与者都熟悉安全处理和处理手术刀和尖锐物体的制度程序. </li>
	<li> 准备下列工具: 剪刀、镊子、手术刀刀片 (No. 15 和 No. 22)、金属探针和金属手术刀柄的钝端 ( 图 1 )。使用手术刀柄钝端, 以尽量减少对微妙的神经纤维的损害, 并保存主要的白质纤维束 ( 图 2 ) 13 。
	<li> 定位大脑, 使其腹面朝向向上. </li>
</ol> 2。解剖过程 注意: 解剖需要大约2到 3 h 来完成 <ol> <li> 用一对创伤 (钝) 钳从两个大脑半球取出蛛网膜和相关的血管. </li>
	<li> 轻轻提起小脑并定位下 colliculi。把手术刀刀片 (No. 15) 附着在一个长的手术刀柄只是尾 colliculi 和轴向通过脑干切割。保持刀片尽可能接近水平, 以避免损害小脑。注意保护中脑的顶. </li>
	<li> 将大脑定位到左侧或右侧的侧裂。从缘回, 使用手术刀柄钝端轻轻地移除表面皮层层。先轻轻向前移动, 然后在侧沟下方, 分别显示在顶叶、额叶和颞裂片上的水平关联纤维束. </li>
	<li> 沿着与上、下纵 fasciculi 连接的脑岛后缘周围的纤维的方向, 以显示弓状束. </li>
	<li> 前, 轻轻地移除中间颞侧和下额回的其余表层皮质层, 以揭示连接颞叶和额部的钩束纤维 </li>
	<li> 识别岛皮层的短回, 然后移除脑岛。然后除去极端的胶囊和 claustrum, 以揭示潜在的外部胶囊。注意状核深部形成的凸出囊。向皮层的背表面移动, 露出日冕的纤维 ( 图 4 ). </li>
	<li> 取出大脑背侧的剩余皮质和底层白质, 以达到扣带回。继续使用钝端的手术刀手柄, 以消除扣带皮层, 以揭示带, 白色物质大片连接前穿孔物质与海马回. </li>
	<li> 使用相同的技术将带从后向前移除, 以揭示胼胝体, 由连接两个大脑半球的合纤维组成。胼胝体 (躯干) 的背部现在将可见 ( 图 6 ). </li>
	<li> 在对侧大脑半球重复步骤2.3 至 2.8. </li>
	<li> 触并确定一个半球的侧脑室的程度。使用探针, 穿刺侧壁的脑室在三角区的位置。使用大小24刀片 (附在一个 No. 4 手术刀手柄) 进入穿刺现场, 并削减下打开整个长度的下角的侧脑室. </li>
	<li> 现在返回到心室侧副三角区, 以将切口向胼胝体的胼胝 (虚线在 图 5 ) 中延伸. </li>
	<li> 在另一个半球上重复步骤2.10 和 2.11. </li>
	<li> 打开侧脑室的身体, 继续切口从三角区 rostrally 使用切开大约 3 cm 平行与胼胝体在两个半球 (虚线在 图 6 ). </li>
	<li> 加入两个平行切口在每个半球 rostrally 水平的膝和尾水平的胼胝的胼胝体。使用钳, 举行在非手, 轻轻地提起胼胝体在胼胝。用一把锋利的小剪刀, 在支配的手举行, 分开胼胝从部下的隔膜隔。一旦你到达身体的侧端, 切割胼胝体并将其移除. </li>
	<li> 将大脑的腹面纵向固定在你非的手掌上, 以稳定枕骨和颞区 (后部)。同时, 用你的优势手牢牢地握住大脑的前端, 把你的手指和拇指放在大脑两侧的状核上. </li>
	<li> 使用轻柔的拉扭动作, 将大脑的前后部分开, 并特别注意保持脉络丛的完好。建议同事在场指导分离, 并在使用手术刀的过程中轻轻地将任何剩余的连接组织分开. </li>
</ol>

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作者声明他们没有利益冲突。

本文件的目的是设计一个解剖指南, 向教师和学生传播, 可用于加强人脑深心室和脑室结构的教学和学习。我们设计了一个 step-by 步指南与伴随的图像, 连同一个视频资源, 可以用来帮助理解的形态学的心室和他们的相关结构。解剖技术本身并不新鲜。纤维解剖曾用于研究小脑解剖14。然而, 我们的研究的新奇之处在传统的解剖方法和现代标注的视频制作的结合。这表明解剖, 尽管?...

许多学生努力理解心室系统的负空间, 位于人脑深处1,2。通常使用的资源可供学生研究脑室提供相对粗糙的表达, 复杂的3D 关系, 这些深脑结构。了解心室系统和相关结构的3D 解剖在神经外科是特别重要的, 因为进入心室系统是测量颅内压力、减压心室的最充分利用的技术之一。系统, 并管理药物3。此外, 医学成像技术的飞速发展, 使得3D 解剖学的解释技能得到了培养。
二维 (2D) 部分的大脑在不同的平面通常用于可视化的深脑结构形成的边界负心室空间4。然而, 仅2D 切片的大脑是不够的, 使学生能够理解的整个范围内的心室3D 结构和该地区的精细细节, 如纤维束连接皮质和皮层下结构5。因此, 教育工作者必须依赖学生自己的能力来计算脑室4的可理解的3D 概念。与空间意识斗争的学生发现创建这个3D 图像非常困难。虽然塑料模型和心室铸型提供了一个3D 的心室系统的表现, 但他们无法证明形成了脑室边界的综合关系。学生常常无意识地删除部分塑料模型, 以进入心室系统, 并了解其相互联系。在这个过程中, 他们经常忽略每个结构的详细的相对位置, 并失去对它们的关系的理解 (例如胼胝体形成的侧脑室的顶部)。
开发新的计算机化教学工具已经解决了其中的一些局限性。但是, 许多这些模型仅限于静态文本和图像, 不利用这些新技术提供的交互性7,8。虽然交互式技术使用户能够旋转3D 计算机模型来研究多个视点, 但这可能会混淆一些用户, 特别是那些发现对结构6具有挑战性的新手。此外, 交互式计算机资源已被证明是不太有效的教学更复杂的解剖结构6。因此, 在神经教育的挑战之一是为学生提供资源, 使他们能够充分形象化的心室和欣赏他们的3D 结构和解剖关系, 包括微妙的联想, 投射,和合的光纤束, 它们与脑室结构2形成复杂的关系。
解剖已被证明是一个优秀的教育方法, 学习解剖学7,8。最近的一项研究提供了学生解剖学习神经的好处的证据。在 2016年, 蕾et al.发现, 在参与解剖的学生中, 神经知识的短期和长期保留能力得到了改善.9。虽然技术的进步继续提高3D 计算机模型的准确性和交互性, 但通过动手解剖获得的知识不能在当前的时间10进行数字复制。
在这项研究中, 我们的目的是产生一个人类大脑的可再生解剖。我们选择了一种纤维解剖方法, 因为它可以保存微妙的纤维束和脑室的灰质结构, 从而更好地定义心室的负空间。
在这里, 我们提出了一个全面的 step-by 步指南, 创造一个 prosection 模型的心室和脑室结构连同一个伴随的训练视频, 用于神经教学和学习。这些资源可以用于教学和学习的神经的大脑由教育工作者和学生。

<table border="0" cellpadding="0" cellspacing="0" width="615">
	<tbody>
		<tr height="20">
			<td height="20" style="height:20px;width:216px;">Scalpel Blade No 15</td>
			<td style="width:113px;">Swann-Morton</td>
			<td style="width:119px;">0205</td>
			<td style="width:167px;">Scalpel blade</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Scalpel Blade No 11</td>
			<td style="width:113px;">Swann-Morton</td>
			<td style="width:119px;">0203</td>
			<td>Scalpel blade</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Scalpel Blade No 24</td>
			<td style="width:113px;">Swann-Morton</td>
			<td style="width:119px;">0211</td>
			<td>Scalpel blade</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Long Scalpel handle No3L</td>
			<td style="width:113px;">Swann-Morton</td>
			<td style="width:119px;">0913</td>
			<td>Scalpel handle</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Short Scalpel handle No4G</td>
			<td style="width:113px;">Swann-Morton</td>
			<td style="width:119px;">0934</td>
			<td>Scalpel handle</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Scissors</td>
			<td style="width:113px;"></td>
			<td style="width:119px;"></td>
			<td>Scissors</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Atraumatic Forceps</td>
			<td style="width:113px;"></td>
			<td style="width:119px;"></td>
			<td>Atraumatic forceps</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Toothed Forceps</td>
			<td style="width:113px;"></td>
			<td style="width:119px;"></td>
			<td>Toothed forceps</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Genelyn Arterial Enhanced</td>
			<td style="width:113px;">GMS Inovations</td>
			<td style="width:119px;">AE-475</td>
			<td>Arterial embalming media</td>
		</tr>
	</tbody>
</table>

exploring deep space - uncovering the anatomy of periventricular structures to reveal the lateral ventricles of the human brain

解剖学学生通常提供 two-dimensional (2D) 部分和图像时, 研究脑室解剖学和学生发现这一挑战。因为心室是位于大脑深处的负空间, 所以了解他们的解剖结构的唯一方法就是欣赏它们的边界。看着这些空间的2D 表示, 在任何一个主要的平面中, 都不会使形成心室边界的所有结构的可视化。因此, 仅使用2D 节就需要学生计算自己的3D 心室空间的心理图像。本研究的目的是开发一种可再生的方法解剖人脑, 创造一个教育资源, 以提高学生对复杂的关系, 心室和脑室结构的理解。为了达到这个目的, 我们创建了一个视频资源, 利用纤维解剖的方法, 以 step-by 的步骤, 揭示了侧和第三脑室与紧密相关的边缘系统和基底节结构。这种方法的优点之一是, 它能够划定的白色物质的大片是很难区分使用其他解剖技术。这段视频附有一份书面协议, 它提供了一个系统的描述过程, 以帮助大脑解剖的再现。该方案为教育工作者和学生提供了宝贵的解剖学教学资源。通过遵循这些指导, 教育工作者可以创造教学资源, 学生可以被引导去产生他们自己的大脑解剖, 作为实践活动。我们建议将该视频指南纳入神经教学, 以提高学生对心室形态学和临床相关性的认识。

这种解剖方法通过将大脑分离成前部和后段 (图 7和 图 8) 来暴露心室系统。后面的部分提供了一个内部的观点, 以三角区的后部和下角可以看到延伸到枕和颞叶分别 (图 8)。在下等或世俗垫铁海马体, 形成它的中间墙壁, 清楚地是可看见的象是毛和穹的下肢。
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Watch this Scientific Journal Video about Exploring Deep Space - Uncovering the Anatomy of Periventricular Structures to Reveal the Lateral Ventricles of the Human Brain at JoVE.com

Exploring Deep Space - Uncovering the Anatomy of Periventricular Structures to Reveal the Lateral Ventricles of the Human Brain

探索深空-揭开人脑侧脑室的脑室结构解剖

本文论证了纤维解剖法在三维空间中有效地揭示了人脑表面白质和脑室结构, 有助于学生对心室形态学的理解。

Anatomy students are typically provided with two-dimensional (2D) sections and images when studying cerebral ventricular anatomy and students find this ...

exploring-deep-space-uncovering-anatomy-periventricular-structures-to

Research

JoVE Journal

Neuroscience

16.5K 次观看.  Australian National University. 本文论证了纤维解剖法在三维空间中有效地揭示了人脑表面白质和脑室结构, 有助于学生对心室形态学的理解。

视频: Exploring Deep Space - Uncovering the Anatomy of Periventricular Structures to Reveal the Lateral Ventricles of the Human Brain

Mouse Models of Periventricular Leukomalacia

We describe a protocol for establishing mouse models of periventricular leukomalacia (PVL). PVL is the predominant form of brain injury in premature infants and the most common antecedent of cerebral palsy. PVL is characterized by periventricular white matter damage with prominent oligodendroglial injury. Hypoxia/ischemia with or without systemic infection/inflammation are the primary causes of PVL. We use P6 mice to create models of neonatal brain injury by the induction of hypoxia/ischemia with or without systemic infection/inflammation with unilateral carotid ligation followed by exposure to hypoxia with or without injection of the endotoxin lipopolysaccharide (LPS). Immunohistochemistry of myelin basic protein (MBP) or O1 and electron microscopic examination show prominent myelin loss in cerebral white matter with additional damage to the hippocampus and thalamus. Establishment of mouse models of PVL will greatly facilitate the study of disease pathogenesis using available transgenic mouse strains, conduction of drug trials in a relatively high throughput manner to identify candidate therapeutic agents, and testing of stem cell transplantation using immunodeficiency mouse strains.

We established mouse models of periventricular leukomalacia (PVL), the predominant brain injury in premature infants characterized by periventricular white matter lesions. Hypoxia/ischemia with/without systemic infection are the primary causes of PVL. Unilateral carotid ligation and hypoxia exposure with/without lipopolysaccharide injection creates PVL-like lesions in P6 mice.

脑室周围白质软化的小鼠模型

We describe a protocol for establishing mouse models of periventricular leukomalacia (PVL). PVL is the predominant form of brain injury in premature ...

科研

神经科学

Morphometric Analyses of Retinal Sections

Morphometric analyses of retinal sections have been used in examining retinal diseases. For examples, neuronal cells were significantly lost in the retinal ganglion cell layer (RGCL) in rat models with N-methyl-D-aspartate (NMDA)&#x2013;induced excitotoxicity1, retinal ischemia-reperfusion injury2 and glaucoma3. Reduction of INL and inner plexiform layer (IPL) thicknesses were reversed with citicoline treatment in rats' eyes subjected to kainic acid-mediated glutamate excitotoxicity4. Alteration of RGC density and soma sizes were observed with different drug treatments in eyes with elevated intraocular pressure3,5,6. Therefore, having objective methods of analyzing the retinal morphometries may be of great significance in evaluating retinal pathologies and the effectiveness of therapeutic strategies.
The retinal structure is multi-layers and several different kinds of neurons exist in the retina. The morphometric parameters of retina such as cell number, cell size and thickness of different layers are more complex than the cell culture system. Early on, these parameters can be detected using other commercial imaging software. The values are normally of relative value, and changing to the precise value may need further accurate calculation. Also, the tracing of the cell size and morphology may not be accurate and sensitive enough for statistic analysis, especially in the chronic glaucoma model. The measurements used in this protocol provided a more precise and easy way. And the absolute length of the line and size of the cell can be reported directly and easy to be copied to other files. For example, we traced the margin of the inner and outer most nuclei in the INL and formed a line then using the software to draw a 90 degree angle to measure the thickness. While without the help of the software, the line maybe oblique and the changing of retinal thickness may not be repeatable among individual observers. In addition, the number and density of RGCs can also be quantified. This protocol successfully decreases the variability in quantitating features of the retina, increases the sensitivity in detecting minimal changes.
 	This video will demonstrate three types of morphometric analyses of the retinal sections. They include measuring the INL thickness, quantifying the number of RGCs and measuring the sizes of RGCs in absolute value. These three analyses are carried out with Stereo Investigator (MBF Bioscience &mdash; MicroBrightField, Inc.). The technique can offer a simple but scientific platform for morphometric analyses.

This video demonstrates three types of morphometric analyses of the retina, which include measuring the inner nuclear layer thickness, quantifying the number of retinal ganglion cells (RGCs) and measuring the sizes of RGCs. The technique can offer a simple but scientific platform for morphometric analyses.

视网膜节的形态分析

Morphometric analyses of retinal sections have been used in examining retinal diseases. For examples, neuronal cells were significantly lost in the ...

Manual Drainage of the Zebrafish Embryonic Brain Ventricles

Cerebrospinal fluid (CSF) is a protein rich fluid contained within the brain ventricles. It is present during early vertebrate embryonic development and persists throughout life. Adult CSF is thought to cushion the brain, remove waste, and carry secreted molecules1,2. In the adult and older embryo, the majority of CSF is made by the choroid plexus, a series of highly vascularized secretory regions located adjacent to the brain ventricles3-5. In zebrafish, the choroid plexus is fully formed at 144 hours post fertilization (hpf)6. Prior to this, in both zebrafish and other vertebrate embryos including mouse, a significant amount of embryonic CSF (eCSF) is present . These data and studies in chick suggest that the neuroepithelium is secretory early in development and may be the major source of eCSF prior to choroid plexus development7.
eCSF contains about three times more protein than adult CSF, suggesting that it may have an important role during development8,9. Studies in chick and mouse demonstrate that secreted factors in the eCSF, fluid pressure, or a combination of these, are important for neurogenesis, gene expression, cell proliferation, and cell survival in the neuroepithelium10-20. Proteomic analyses of human, rat, mouse, and chick eCSF have identified many proteins that may be necessary for CSF function. These include extracellular matrix components, apolipoproteins, osmotic pressure regulating proteins, and proteins involved in cell death and proliferation21-24. However, the complex functions of the eCSF are largely unknown.
We have developed a method for removing eCSF from zebrafish brain ventricles, thus allowing for identification of eCSF components and for analysis of the eCSF requirement during development. Although more eCSF can be collected from other vertebrate systems with larger embryos, eCSF can be collected from the earliest stages of zebrafish development, and under genetic or environmental conditions that lead to abnormal brain ventricle volume or morphology. Removal and collection of eCSF allows for mass spectrometric analysis, investigation of eCSF function, and reintroduction of select factors into the ventricles to assay their function. Thus the accessibility of the early zebrafish embryo allows for detailed analysis of eCSF function during development.

We present a method to collect cerebrospinal fluid (CSF) and to create a system which lacks CSF within the embryonic zebrafish brain ventricular system. This allows for further examination of CSF composition and its requirement during embryonic brain development.

手动排水的斑马鱼胚胎脑室

Cerebrospinal fluid  (CSF) is a protein rich fluid contained within the brain ventricles. It is  present during early vertebrate embryonic development and ...

3D Modeling of the Lateral Ventricles and Histological Characterization of Periventricular Tissue in Humans and Mouse

The ventricular system carries and circulates cerebral spinal fluid (CSF) and facilitates clearance of solutes and toxins from the brain. The functional units of the ventricles are ciliated epithelial cells termed ependymal cells, which line the ventricles and through ciliary action are capable of generating laminar flow of CSF at the ventricle surface. This monolayer of ependymal cells also provides barrier and filtration functions that promote exchange between brain interstitial fluids (ISF) and circulating CSF. Biochemical changes in the brain are thereby reflected in the composition of the CSF and destruction of the ependyma can disrupt the delicate balance of CSF and ISF exchange. In humans there is a strong correlation between lateral ventricle expansion and aging. Age-associated ventriculomegaly can occur even in the absence of dementia or obstruction of CSF flow. The exact cause and progression of ventriculomegaly is often unknown; however, enlarged ventricles can show regional and, often, extensive loss of ependymal cell coverage with ventricle surface astrogliosis and associated periventricular edema replacing the functional ependymal cell monolayer. Using MRI scans together with postmortem human brain tissue, we describe how to prepare, image and compile 3D renderings of lateral ventricle volumes, calculate lateral ventricle volumes, and characterize periventricular tissue through immunohistochemical analysis of en face lateral ventricle wall tissue preparations. Corresponding analyses of mouse brain tissue are also presented supporting the use of mouse models as a means to evaluate changes to the lateral ventricles and periventricular tissue found in human aging and disease. Together, these protocols allow investigations into the cause and effect of ventriculomegaly and highlight techniques to study ventricular system health and its important barrier and filtration functions within the brain.

Using MRI scans (human), 3D imaging software, and immunohistological analysis, we document changes to the brain&#8217;s lateral ventricles. Longitudinal 3D mapping of lateral ventricle volume changes and characterization of periventricular cellular changes that occur in the human brain due to aging or disease are then modeled in mice.

脑室周围组织的侧脑室及组织学特性对人类和鼠标3D建模

The ventricular system carries and circulates cerebral spinal fluid (CSF) and facilitates clearance of solutes and toxins from the brain. The functional ...

Analysis of Gene Expression Changes in the Rat Hippocampus After Deep Brain Stimulation of the Anterior Thalamic Nucleus

Deep brain stimulation (DBS) surgery, targeting various regions of the brain such as the basal ganglia, thalamus, and subthalamic regions, is an effective treatment for several movement disorders that have failed to respond to medication. Recent progress in the field of DBS surgery has begun to extend the application of this surgical technique to other conditions as diverse as morbid obesity, depression and obsessive compulsive disorder. Despite these expanding indications, little is known about the underlying physiological mechanisms that facilitate the beneficial effects of DBS surgery. One approach to this question is to perform gene expression analysis in neurons that receive the electrical stimulation. Previous studies have shown that neurogenesis in the rat dentate gyrus is elicited in DBS targeting of the anterior nucleus of the thalamus1. DBS surgery targeting the ATN is used widely for treatment refractory epilepsy. It is thus of much interest for us to explore the transcriptional changes induced by electrically stimulating the ATN. In this manuscript, we describe our methodologies for stereotactically-guided DBS surgery targeting the ATN in adult male Wistar rats. We also discuss the subsequent steps for tissue dissection, RNA isolation, cDNA preparation and quantitative RT-PCR for measuring gene expression changes. This method could be applied and modified for stimulating the basal ganglia and other regions of the brain commonly clinically targeted. The gene expression study described here assumes a candidate target gene approach for discovering molecular players that could be directing the mechanism for DBS.

The mechanism underlying the therapeutic effects of Deep Brain Stimulation (DBS) surgery needs investigation. The methods presented in this manuscript describe an experimental approach to examine the cellular events triggered by DBS by analyzing the gene expression profile of candidate genes that can facilitate neurogenesis post DBS surgery.

基因表达的变化大鼠海马后，前丘脑核脑深部刺激分析

Deep brain stimulation (DBS) surgery, targeting various regions of the brain such as the basal ganglia, thalamus, and subthalamic regions, is an effective ...

Investigating the Function of Deep Cortical and Subcortical Structures Using Stereotactic Electroencephalography: Lessons from the Anterior Cingulate Cortex

Stereotactic Electroencephalography (SEEG) is a technique used to localize seizure foci in patients with medically intractable epilepsy. This procedure involves the chronic placement of multiple depth electrodes into regions of the brain typically inaccessible via subdural grid electrode placement. SEEG thus provides a unique opportunity to investigate brain function. In this paper we demonstrate how SEEG can be used to investigate the role of the dorsal anterior cingulate cortex (dACC) in cognitive control. We include a description of the SEEG procedure, demonstrating the surgical placement of the electrodes. We describe the components and process required to record local field potential (LFP) data from consenting subjects while they are engaged in a behavioral task. In the example provided, subjects play a cognitive interference task, and we demonstrate how signals are recorded and analyzed from electrodes in the dorsal anterior cingulate cortex, an area intimately involved in decision-making. We conclude with further suggestions of ways in which this method can be used for investigating human cognitive processes.

Stereotactic Electroencephalography (SEEG) is an operative technique used in epilepsy surgery to help localize seizure foci. It also affords a unique opportunity to investigate brain function. Here we describe how SEEG can be used to investigate cognitive processes in human subjects.

调查深皮层的功​​能和皮层下结构采用立体定向脑电图:由前扣带皮层的经验教训

Stereotactic Electroencephalography (SEEG) is a technique used to localize seizure foci in patients with medically intractable epilepsy. This procedure ...

Live Imaging of the Ependymal Cilia in the Lateral Ventricles of the Mouse Brain

Multiciliated ependymal cells line the ventricles in the adult brain. Abnormal function or structure of ependymal cilia is associated with various neurological deficits. The current ex vivo live imaging of motile ependymal cilia technique allows for a detailed study of ciliary dynamics following several steps. These steps include: mice euthanasia with carbon dioxide according to protocols of The University of Toledo&#8217;s Institutional Animal Care and Use Committee (IACUC); craniectomy followed by brain removal and sagittal brain dissection with a vibratome or sharp blade to obtain very thin sections through the brain lateral ventricles, where the ependymal cilia can be visualized. Incubation of the brain&#8217;s slices in a customized glass-bottom plate containing Dulbecco&#8217;s Modified Eagle&#8217;s Medium (DMEM)/High-Glucose at 37 &#176;C in the presence of 95%/5% O2/CO2 mixture is essential to keep the tissue alive during the experiment. A video of the cilia beating is then recorded using a high-resolution differential interference contrast microscope. The video is then analyzed frame by frame to calculate the ciliary beating frequency. This allows distinct classification of the ependymal cells into three categories or types based on their ciliary beating frequency and angle. Furthermore, this technique allows the use of high-speed fluorescence imaging analysis to characterize the unique intracellular calcium oscillation properties of ependymal cells as well as the effect of pharmacological agents on the calcium oscillations and the ciliary beating frequency. In addition, this technique is suitable for immunofluorescence imaging for ciliary structure and ciliary protein localization studies. This is particularly important in disease diagnosis and phenotype studies. The main limitation of the technique is attributed to the decrease in live motile cilia movement as the brain tissue starts to die.

Using high-resolution differential interference contrast (DIC) microscopy, an ex vivo observation of the beating of motile ependymal cilia located within the mouse brain ventricles is demonstrated by live-imaging. The technique allows a recording of the unique ciliary beating frequency and beating angle as well as their intracellular calcium oscillation pacing properties.

室管膜纤毛在小鼠脑侧脑室实时成像

Multiciliated ependymal cells line the ventricles in the adult brain. Abnormal function or structure of ependymal cilia is associated with various ...

A Simple One-step Dissection Protocol for Whole-mount Preparation of Adult Drosophila Brains

There is an increasing interest in using Drosophila to model human brain degenerative diseases, map neuronal circuitries in adult brains, and study the molecular and cellular basis of higher brain functions. A whole-mount preparation of adult brains with well-preserved morphology is critical for such whole brain-based studies, but can be technically challenging and time-consuming. This protocol describes an easy-to-learn, one-step dissection approach of an adult fly head in less than 10 s, while keeping the intact brain attached to the rest of the body to facilitate subsequent processing steps. The procedure helps remove most of the eye and tracheal tissues normally associated with the brain that can interfere with the later imaging step, and also places less demand on the quality of the dissecting forceps. Additionally, we describe a simple method that allows convenient flipping of the mounted brain samples on a coverslip, which is important for imaging both sides of the brains with similar signal intensity and quality. As an example of the protocol, we present an analysis of dopaminergic (DA) neurons in adult brains of WT (w1118) flies. The high efficacy of the dissection method makes it particularly useful for large-scale adult brain-based studies in Drosophila.

A Simple One-step Dissection Protocol for Whole-mount Preparation of Adult Drosophila Brains

The adult Drosophila brain is a valuable system for studying neuronal circuitry, higher brain functions, and complex disorders. An efficient method to dissect whole brain tissue from the small fly head will facilitate brain-based studies. Here we describe a simple, one-step dissection protocol of adult brains with well-preserved morphology.

一个简单的一步解剖协议成人全安装准备<em&gt;果蝇</em&gt;脑

There is an increasing interest in using Drosophila to model human brain degenerative diseases, map neuronal circuitries in adult brains, and study the ...

Sublimation of DAN Matrix for the Detection and Visualization of Gangliosides in Rat Brain Tissue for MALDI Imaging Mass Spectrometry

Sample preparation is key for optimal detection and visualization of analytes in Matrix-assisted Laser Desorption/Ionization (MALDI) Imaging Mass Spectrometry (IMS) experiments. Determining the appropriate protocol to follow throughout the sample preparation process can be difficult as each step must be optimized to comply with the unique characteristics of the analytes of interest. This process involves not only finding a compatible matrix that can desorb and ionize the molecules of interest efficiently, but also selecting the appropriate matrix deposition technique. For example, a wet matrix deposition technique, which entails dissolving a matrix in solvent, is superior for desorption of most proteins and peptides, whereas dry matrix deposition techniques are particularly effective for ionization of lipids. Sublimation has been reported as a highly efficient method of dry matrix deposition for the detection of lipids in tissue by MALDI IMS due to the homogeneity of matrix crystal deposition and minimal analyte delocalization as compared to many wet deposition methods 1,2. Broadly, it involves placing a sample and powdered matrix in a vacuum-sealed chamber with the samples pressed against a cold surface. The apparatus is then lowered into a heated bath (sand or oil), resulting in sublimation of the powdered matrix onto the cooled tissue sample surface. Here we describe a sublimation protocol using 1,5-diaminonaphthalene (DAN) matrix for the detection and visualization of gangliosides in the rat brain using MALDI IMS.

A protocol for the sublimation of DAN matrix onto rat brain tissue for the detection of gangliosides using MALDI Imaging Mass Spectrometry is presented.

DAN矩阵升华为神经节苷脂对大鼠脑组织的检测和可视化MALDI成像质谱

Sample preparation is key for optimal detection and visualization of analytes in Matrix-assisted Laser Desorption/Ionization (MALDI) Imaging Mass ...

Recording Synaptic Plasticity in Acute Hippocampal Slices Maintained in a Small-volume Recycling-, Perfusion-, and Submersion-type Chamber System

Even though experiments on brain slices have been in use since 1951, problems remain that reduce the probability of achieving a stable and successful analysis of synaptic transmission modulation when performing field potential or intracellular recordings. This manuscript describes methodological aspects that might be helpful in improving experimental conditions for the maintenance of acute brain slices and for recording field excitatory postsynaptic potentials in a commercially available submersion chamber with an outflow-carbogenation unit. The outflow-carbogenation helps to stabilize the oxygen level in experiments that rely on the recycling of a small buffer reservoir to enhance the cost-efficiency of drug experiments. In addition, the manuscript presents representative experiments that examine the effects of different carbogenation modes and stimulation paradigms on the activity-dependent synaptic plasticity of synaptic transmission.

This protocol describes the stabilization of the oxygen level in a small volume of recycled buffer and methodological aspects of recording activity-dependent synaptic plasticity in submerged acute hippocampal slices.

小容积回收-灌注-浸没式室系统中维持的急性海马切片的突触可塑性记录

Even though experiments on brain slices have been in use since 1951, problems remain that reduce the probability of achieving a stable and successful ...