作者要感谢卡罗琳·哈特利和 Rebeccah 斯莱特 (英国牛津大学儿科系) 对我们的论文和沃尔特 Magerl (生物医学和医学技术中心神经生理学系) 的评论。曼海姆 (CBTM), 德国海德堡大学) 支持我们的技术设备和知识。

这项研究得到了瑞士西北部主管道德委员会 (EKNZ 2015-079) 的批准, 并在测量前获得了有关参与者的书面父母同意。
1. 准备
<ol><li>确保孩子已经安顿好了。它应该是安静和安定的, 不用吮运动在录音期间由于运动文物。婴儿可以睡着9。</li>
	<li>用卷尺测量新生儿头部周长, 确定脑电图帽的大小。</li>
	<li>确定活动电极位置: Cz/顶点位置通过标记中间点之间的 nasion 和 inion 和中间点之间的左, 右 preaurical 点与皮肤标记铅笔。为定位电极也请参见图 1。</li>
	<li>确定地面电极的位置: 右前额 (Fp2) 和参考电极: 左乳突 (A1)。</li>
	<li>用棉签清洗电极部位 (Cz、Fp2 和 A1)。在大量的头发的情况下, 分开的头发, 以可视化头皮。</li>
	<li>用棉签用棉签轻轻擦洗电极部位, 以降低阻抗。</li>
	<li>将脑电图帽与附着在新生儿头上的电极放在一起。</li>
	<li>使用带有短塑料针头的注射器向电极注入导电脑电凝胶, 以优化电极和头皮之间的接触。</li>
	<li>调整电极, 直到阻抗低于50ω, 如制造商的建议和发布的准则12所述。</li>
	<li>放置一个摄像头来记录新生儿的面部表情。</li>
	<li>将平尖探头连接到触点触发装置, 固定在 EEG 记录装置上。当平尖探针到达皮肤的标称力时, 触发信号由触点触发装置产生。这个信号被发送到计算机, 用一个触发标记标记脑电图记录。</li>
</ol>2. 测量
<ol><li>选择一个研究名称来存储数据。</li>
	<li>
添加在线带通滤波器:
	<ol>
<li>显示筛选器: 低截止滤波器: 频率: 1 赫兹 高截止滤波器: 频率:70 赫兹 凹槽过滤器:50 赫兹 采样率: 2000 赫兹</li>
	</ol>
</li>
	<li>启动脑电图和录像</li>
	<li>将新生儿的右手保持在水平位置。刺激也可以应用在不同的站点上,例如脚, 这将导致比手更长的滞后时间, 请参阅详细信息3.7。</li>
	<li>第一记录背景脑电图活动。当新生儿的手举行, 注释的脑电图记录手动记录期间, 没有刺激的应用和婴儿休息。</li>
	<li>根据新生儿右手的研究设计, 进行所需量的平尖探针刺激。在我们的实验设计中, 设置了50平尖端探针刺激的数量。小心地使用平尖端探针垂直地对新生儿的手, 因此提示不弯曲和正确的力量被应用。使用最小2到3秒的 inter-stimulus 间隔 (ISI)6执行平面尖端探头刺激, 以避免求和。</li>
	<li>停止所有的录音。</li>
	<li>记录实验设置细节。</li>
</ol>3. 数据分析
<ol><li>使用高通滤波器在1赫兹和低通滤波器的30赫兹下离线过滤原始脑电图数据。</li>
	<li>片段数据在世纪1500毫秒 (500 毫秒之前到1000毫秒在刺激开始之后)。</li>
	<li>对 pre-stimulus 间隔执行基线校正。</li>
	<li>在目视检查后, 手动拒绝包含人工制品的 EEG 世纪, 如机芯和噪音。请参阅教程大纲: https://sccn.ucsd.edu/wiki/Chapter_01:_Rejecting_Artifacts</li>
	<li>平均 EEG 世纪 (为背景和刺激反应分开)。</li>
	<li>木质过滤器数据以最大抖动±50毫秒在时间窗口 0-1000 毫秒后刺激开始。这使得婴儿之间的潜伏期差异。这样做的背景和刺激反应分开。</li>
	<li>项目一个有害诱发脑活动的模板, 这是定义在一个独立的数据集, 并已详细描述了其他地方11的数据, 以确定在每个单独的试验中的有害诱发反应的程度。该模板描述了有害诱发反应的特征波形, 震级反映了在个体试验中这种有害诱发反应的数量。 注意: 模板投射的时间窗口取决于婴儿的年龄和刺激的位置。如果刺激适用于手然后时间窗口的兴趣是 200-500 毫秒后刺激11。如果刺激被应用到脚然后时间窗口兴趣是 400-700 ms 岗位刺激11。请注意, 该模板目前仅在 34-43 周的妊娠期内进行了验证。有关使用此模板的详细方法以及有关限制的讨论, 请参见哈特利et al. 201711。</li>
	<li>检查使用该模板计算的有害诱发脑活动的幅度在刺激后明显高于背景脑电图。</li>
	<li>使用新生儿面部编码系统2,13, 事后分析行为面部表情 (摄像机与 EEG 记录相关)。</li>
</ol>

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作者没有什么可透露的。

这里介绍的方法显示了如何使用脑电图记录和平尖探针刺激对新生儿的有害诱发脑活动进行客观的测量, 以应用实验性的有害刺激。这项技术可用于各种临床设置, 以检测伤害,例如在非语言的人, 如新生儿。完整的研究可以在15分钟内完成, 包括放置婴儿, 识别, 准备和安装电极, 最后应用和记录50有害刺激。这种刺激不会引起婴儿的行为窘迫, 这一方法为评估婴儿的伤害提供了客观的方法。...

疼痛是一种不愉快的感觉和情感体验, 与实际或潜在的组织损害有关, 或在这种损害方面描述为1。无法口头交流并不能否定个人正在经历痛苦的可能性, 但却使评估减轻疼痛的治疗具有挑战性, 例如新生儿2。一些行为和生理指标被用来评估非语言患者的疼痛。不同的尺度已经发展了多年, 选择取决于类型的刺激, 孕龄和环境中的新生儿是嵌入式3,4,5。这些疼痛评估工具要么依赖于评分员的解释, 要么需要二级生理指标。
在这个视频中, 我们提出了一种方法来评估伤害的电生理记录优化用于新生儿。伤害被定义为有害刺激编码的神经过程。因此, 量化伤害是一个优雅和客观的方法来确定神经输入的非语言的人。此外, 脑电图 (eeg) 检测到的皮层活动与有害事件强度相关,5,6。
这里提出的方法结合脑电图记录与机械刺激产生的有害刺激与扁平尖端探针, 也称为针刺7, 这不是皮肤破损, 并没有造成行为困扰6。它已经表明, 伤害以下的加标点刺激主要是介导的 Aδ纤维, 不需要皮肤破裂病变8和伤害性特定电位的大小不依赖于睡眠状态9。在与新生儿的研究环境中应用这种方法时, 家长也很好地接受了这一探针。探针是电子连接到 eeg 记录系统, 使脑电图记录准确标签时, 探针接触皮肤。这大大简化了时间锁定的过程, 是所有后续脑电图分析的前提。为了最大限度地减少婴儿脑电图记录的准备时间, 我们使用了一个改进的国际10/20 电极放置系统, 我们减少了电极的数量, 以最低要求的三电极 (图 1)。中心顶点 Cz 电极, 其中有害诱发脑活动是最大的9,10,11, 与一个参考和一个接地电极一起使用。

<table>
	<tbody>
		<tr>
			<td>Easycap</td>
			<td>EASYCAP GmbH</td>
			<td>AC-32-C</td>
			<td>EEG caps for infants sizes 34 and 36</td>
		</tr>
		<tr>
			<td>actiCAP</td>
			<td>Brain Products GmbH</td>
			<td>BP-04243-SIG</td>
			<td>active electrodes</td>
		</tr>
		<tr>
			<td>ImpBox</td>
			<td>Brain Products GmbH</td>
			<td></td>
			<td>impedance measurement</td>
		</tr>
		<tr>
			<td>V-Amp</td>
			<td>Brain Products GmbH</td>
			<td></td>
			<td>EEG recording device</td>
		</tr>
		<tr>
			<td>Contact trigger for pinprick stimulation</td>
			<td>MRC Systems GmbH</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>PinPrick stimulator set</td>
			<td>MRC Systems GmbH</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>EEG prepping paste</td>
			<td>USB Pharmacy</td>
			<td></td>
			<td>contains sodium chloride, pumice stone, propylene glycol</td>
		</tr>
		<tr>
			<td>SuperVisc</td>
			<td>EASYCAP GmbH</td>
			<td></td>
			<td>Electrolyte-Gel for active electrodes</td>
		</tr>
		<tr>
			<td>Brain Vision Recorder</td>
			<td>Brain Products GmbH</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Brain Vision Analyzer</td>
			<td>Brain Products GmbH</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>MATLAB using EEGLAB
			
			
			
			
			
			</td>
			<td>Swartz Center for Computational Neuroscience, University of California San Diego</td>
			<td></td>
			<td>For EEG processing, including averaging of all EEG epochs
			
			
			
			</td>
		</tr>
	</tbody>
</table>

electrophysiological measurement of noxious-evoked brain activity in neonates using a flat-tip probe coupled to electroencephalography

疼痛是一种不愉快的感官和情感体验。在非语言患者中, 即使用疼痛评估工具也很难测量疼痛。这些工具是主观的或确定的次要生理指标, 也有一定的限制, 特别是在探索的有效性镇痛。由于皮质加工对疼痛知觉至关重要, 脑活动措施可为评估婴儿疼痛提供一种有用的方法。在这里, 我们提出了一种方法来评估伤害与电生理脑活动录音优化用于新生儿。为了产生高度标准化和可重复性的有害刺激, 我们应用了机械刺激与平尖端探针,例如, 针刺, 这不是皮肤破损, 不会造成行为困扰。有害诱发电位可客观测量非言语患者的伤害。这种方法可用于新生儿早在34周的孕龄。此外, 它可以应用在不同的情况下, 如测量镇痛或麻醉药物的功效。

图 1是使用修改过的国际10/20 电极放置系统的电极定位的图解表示。图 2显示了在使用带有 32 mN 力的平头探针进行一项单一有害刺激的应用之前和之后的脑电图活动, 这种刺激发生在新生儿的右手协议 (图 2A) 中。在大约 300 ms 后刺激开始 (时间 0), 毒性诱发反应是可见的。图 2B显示了50...

Watch this Scientific Journal Video about Electrophysiological Measurement of Noxious-evoked Brain Activity in Neonates Using a Flat-tip Probe Coupled to Electroencephalography at JoVE.com

Electrophysiological Measurement of Noxious-evoked Brain Activity in Neonates Using a Flat-tip Probe Coupled to Electroencephalography

应用平板探针结合脑电图对新生儿有害诱发脑活动的电生理测量

测量非语言患者的疼痛是一个挑战。在本研究中, 我们结合脑电记录与刺激使用平尖端探针检测有害诱发脑活动的客观方式。

Pain is an unpleasant sensory and emotional experience. In non-verbal patients, it is very difficult to measure pain, even with pain assessment tools. ...

electrophysiological-measurement-noxious-evoked-brain-activity

Research

JoVE Journal

Medicine

6.5K 次观看.  University of Basel Children's Hospital. 测量非语言患者的疼痛是一个挑战。在本研究中, 我们结合脑电记录与刺激使用平尖端探针检测有害诱发脑活动的客观方式。

视频: Electrophysiological Measurement of Noxious-evoked Brain Activity in Neonates Using a Flat-tip Probe Coupled to Electroencephalography

Segmentation and Measurement of Fat Volumes in Murine Obesity Models Using X-ray Computed Tomography

Obesity is associated with increased morbidity and mortality as well as reduced metrics in quality of life.1 Both environmental and genetic factors are associated with obesity, though the precise underlying mechanisms that contribute to the disease are currently being delineated.2,3 Several small animal models of obesity have been developed and are employed in a variety of studies.4 A critical component to these experiments involves the collection of regional and/or total animal fat content data under varied conditions.
Traditional experimental methods available for measuring fat content in small animal models of obesity include invasive (e.g. ex vivo measurement of fat deposits) and non-invasive (e.g. Dual Energy X-ray Absorptiometry (DEXA), or Magnetic Resonance (MR)) protocols, each of which presents relative trade-offs. Current invasive methods for measuring fat content may provide details for organ and region specific fat distribution, but sacrificing the subjects will preclude longitudinal assessments. Conversely, current non-invasive strategies provide limited details for organ and region specific fat distribution, but enable valuable longitudinal assessment. With the advent of dedicated small animal X-ray computed tomography (CT) systems and customized analytical procedures, both organ and region specific analysis of fat distribution and longitudinal profiling may be possible. Recent reports have validated the use of CT for in vivo longitudinal imaging of adiposity in living mice.5,6 Here we provide a modified method that allows for fat/total volume measurement, analysis and visualization utilizing the Carestream Molecular Imaging Albira CT system in conjunction with PMOD and Volview software packages.

Fat content analysis is routinely conducted in studies utilizing murine obesity models. Emerging methods in small animal CT imaging and analysis are providing for longitudinal detail rich fat content analysis. Here we detail step by step procedures for performing small animal CT imaging, analysis, and visualization.

利用X射线计算机断层扫描在小鼠肥胖模型的FAT卷的分割和测量

Obesity is associated with increased morbidity and mortality as well as reduced metrics in quality of life.1 Both environmental and genetic factors are ...

科研

医学

Epidural Intracranial Pressure Measurement in Rats Using a Fiber-optic Pressure Transducer

Elevated intracranial pressure (ICP) is a significant problem in several forms of ischemic brain injury including stroke, traumatic brain injury and cardiac arrest. This elevation may result in further neurological injury, in the form of transtentorial herniation1,2,3,4, midbrain compression, neurological deficit or increased cerebral infarct2,4. Current therapies are often inadequate to control elevated ICP in the clinical setting5,6,7 . Thus there is a need for accurate methods of ICP measurement in animal models to further our understanding of the basic mechanisms and to develop new treatments for elevated ICP.
In both the clinical and experimental setting ICP cannot be estimated without direct measurement. Several methods of ICP catheter insertion currently exist. Of these the intraventricular catheter has become the clinical 'gold standard' of ICP measurement in humans8. This method involves the partial removal of skull and the instrumentation of the catheter through brain tissue. Consequently, intraventricular catheters have an infection rate of 6-11%9. For this reason, subdural and epidural cannulations have become the preferred methods in animal models of ischemic injury. 
Various ICP measurement techniques have been adapted for animal models, and of these, fluid-filled telemetry catheters10 and solid state catheters are the most frequently used11,12,13,14,15. The fluid-filled systems are prone to developing air bubbles in the line, resulting in false ICP readings. Solid state probes avoid this problem (Figure 1). An additional problem is fitting catheters under the skull or into the ventricles without causing any brain injury that might alter the experimental outcomes. Therefore, we have developed a method that places an ICP catheter contiguous with the epidural space, but avoids the need to insert it between skull and brain. 
An optic fibre pressure catheter (420LP, SAMBA Sensors, Sweden) was used to measure ICP at the epidural location because the location of the pressure sensor (at the very tip of the catheter) was found to produce a high fidelity ICP signal in this model. There are other manufacturers of similar optic fibre technologies13 that may be used with our methodology. Alternative solid state catheters, which have the pressure sensor located at the side of the catheter tip, would not be appropriate for this model as the signal would be dampened by the presence of the monitoring screw. 
Here, we present a relatively simple and accurate method to measure ICP. This method can be used across a wide range of ICP related animal models.

A novel technique to record the pressures within the skull is described. The minimally invasive method uses a fibre-optic pressure sensing system to accurately measure intracranial pressure (ICP) in anaesthetized rats without causing significant brain trauma. The technique may be used in a wide range of experimental models.

硬膜外颅内压测量中使用的光纤压力传感器的大鼠

Elevated intracranial pressure (ICP) is a significant problem in several forms of ischemic brain injury including stroke, traumatic brain injury and ...

The Measurement and Treatment of Suppression in Amblyopia

Amblyopia, a developmental disorder of the visual cortex, is one of the leading causes of visual dysfunction in the working age population. Current estimates put the prevalence of amblyopia at approximately 1-3%1-3, the majority of cases being monocular2. Amblyopia is most frequently caused by ocular misalignment (strabismus), blur induced by unequal refractive error (anisometropia), and in some cases by form deprivation.
Although amblyopia is initially caused by abnormal visual input in infancy, once established, the visual deficit often remains when normal visual input has been restored using surgery and/or refractive correction. This is because amblyopia is the result of abnormal visual cortex development rather than a problem with the amblyopic eye itself4,5 . Amblyopia is characterized by both monocular and binocular deficits6,7 which include impaired visual acuity and poor or absent stereopsis respectively. The visual dysfunction in amblyopia is often associated with a strong suppression of the inputs from the amblyopic eye under binocular viewing conditions8. Recent work has indicated that suppression may play a central role in both the monocular and binocular deficits associated with amblyopia9,10 .
Current clinical tests for suppression tend to verify the presence or absence of suppression rather than giving a quantitative measurement of the degree of suppression. Here we describe a technique for measuring amblyopic suppression with a compact, portable device11,12 . The device consists of a laptop computer connected to a pair of virtual reality goggles. The novelty of the technique lies in the way we present visual stimuli to measure suppression. Stimuli are shown to the amblyopic eye at high contrast while the contrast of the stimuli shown to the non-amblyopic eye are varied. Patients perform a simple signal/noise task that allows for a precise measurement of the strength of excitatory binocular interactions. The contrast offset at which neither eye has a performance advantage is a measure of the "balance point" and is a direct measure of suppression. This technique has been validated psychophysically both in control13,14 and patient6,9,11 populations.
In addition to measuring suppression this technique also forms the basis of a novel form of treatment to decrease suppression over time and improve binocular and often monocular function in adult patients with amblyopia12,15,16 . This new treatment approach can be deployed either on the goggle system described above or on a specially modified iPod touch device15.

Amblyopia is a developmental disorder of the visual cortex that is often accompanied by strong suppression of one eye. We present a new technique for measuring and treating interocular suppression in patients with amblyopia that can be deployed using virtual reality goggles or a portable iPod Touch device.

测量和制止在弱视治疗

Amblyopia, a developmental disorder of the visual cortex, is one of the leading causes of visual dysfunction in the working age population. Current ...

State of the Art Cranial Ultrasound Imaging in Neonates

Cranial ultrasound (CUS) is a reputable tool for brain imaging in critically ill neonates. It is safe, relatively cheap and easy to use, even when a patient is unstable. In addition it is radiation-free and allows serial imaging. CUS possibilities have steadily expanded. However, in many neonatal intensive care units, these possibilities are not optimally used. We present a comprehensive approach for neonatal CUS, focusing on optimal settings, different probes, multiple acoustic windows and Doppler techniques. This approach is suited for both routine clinical practice and research purposes. In a live demonstration, we show how this technique is performed in the neonatal intensive care unit. Using optimal settings and probes allows for better imaging quality and improves the diagnostic value of CUS in experienced hands. Traditionally, images are obtained through the anterior fontanel. Use of supplemental acoustic windows (lambdoid, mastoid, and lateral fontanels) improves detection of brain injury. Adding Doppler studies allows screening of patency of large intracranial arteries and veins. Flow velocities and indices can be obtained. Doppler CUS offers the possibility of detecting cerebral sinovenous thrombosis at an early stage, creating a window for therapeutic intervention prior to thrombosis-induced tissue damage. Equipment, data storage and safety aspects are also addressed.

Cranial ultrasound (CUS) is a valuable tool for brain imaging in critically ill neonates. This video shows a comprehensive approach for neonatal (Doppler) CUS for both clinical and research purposes, including a bedside demonstration of the technique.

艺术颅超声成像在新生儿的状态

Cranial ultrasound (CUS) is a reputable tool for brain imaging in critically ill neonates. It is safe, relatively cheap and easy to use, even when a ...

Molecular Probe Optimization to Determine Cell Mortality in a Photosynthetic Organism (Microcystis aeruginosa) Using Flow Cytometry

Microbial subpopulations in field and laboratory studies have been shown to display high heterogeneity in morphological and physiological parameters. Determining the real time state of a microbial cell goes beyond live or dead categories, as microbes can exist in a dormant state, whereby cell division and metabolic activities are reduced. Given the need for detection and quantification of microbes, flow cytometry (FCM) with molecular probes provides a rapid and accurate method to help determine overall population viability. By using SYTOX Green and SYTOX Orange in the model cyanobacteria Microcystis aeruginosa to detect membrane integrity, we develop a transferable method for rapid indication of single cell mortality. The molecular probes used within this journal will be referred to as green or orange nucleic acid probes respectively (although there are other products with similar excitation and emission wavelengths that have a comparable modes of action, we specifically refer to the fore mentioned probes). Protocols using molecular probes vary between species, differing principally in concentration and incubation times. Following this protocol set out on M.aeruginosa the green nucleic acid probe was optimized at concentrations of 0.5 &#181;M after 30 min of incubation and the orange nucleic acid probe at 1 &#181;M after 10 min. In both probes concentrations less than the stated optimal led to an under reporting of cells with membrane damage. Conversely, 5 &#181;M concentrations and higher in both probes exhibited a type of non-specific staining, whereby &#39;live&#39; cells produced a target fluorescence, leading to an over representation of &#39;non-viable&#39; cell numbers. The positive controls (heat-killed) provided testable dead biomass, although the appropriateness of control generation remains subject to debate. By demonstrating a logical sequence of steps for optimizing the green and orange nucleic acid probes we demonstrate how to create a protocol that can be used to analyse cyanobacterial physiological state effectively.

Molecular Probe Optimization to Determine Cell Mortality in a Photosynthetic Organism Microcystis aeruginosa Using Flow Cytometry

Microbial populations contain substantial cell heterogeneity, which can dictate overall behavior. Molecular probe analysis through flow cytometry can determine physiological states of cells, however its application varies between species. This study provides a protocol to accurately determine cell mortality within a cyanobacterium population, without underestimating or recording false positive results.

分子探针的优化，以确定细胞死亡的光合生物（<em&gt;铜绿微囊藻</em&gt;）使用流式细胞仪

Microbial subpopulations in field and laboratory studies have been shown to display high heterogeneity in morphological and physiological parameters. ...

Transcutaneous Microcirculatory Imaging in Preterm Neonates

Microcirculatory imaging (MI) is a relatively new research tool mainly used in the intensive care setting. MI provides a clear view of the smallest capillaries, arterioles and venules. The magnifying effect visualizes the flow pattern of erythrocytes through these vessels.
It's non-invasive character makes it suitable to apply in (preterm) neonates, even in cardiorespiratory unstable patients. In adults and children, MI is mainly performed sublingually, but this is not possible in preterm infants as these cannot cooperate and the size of the probe is problematic. In preterm infants, MI is therefore performed transcutaneously. Their thin skin makes it possible to obtain high quality images of peripheral microcirculation. 
In this manuscript we will demonstrate the method of transcutaneous MI in preterm infants. We will focus on the different techniques and provide tips to optimize image quality. The highlights of software settings, safety and offline analysis are also addressed.

Microcirculatory imaging (MI) is used to monitor peripheral perfusion in critically ill or preterm neonates. This manuscript and video demonstrates the optimal approach for obtaining high-quality images.

经皮微循环成像在早产儿

Microcirculatory imaging (MI) is a relatively new research tool mainly used in the intensive care setting. MI provides a clear view of the smallest ...

Using Saccadometry with Deep Brain Stimulation to Study Normal and Pathological Brain Function

The oculomotor system involves a large number of brain areas including parts of the basal ganglia, and various neurodegenerative diseases including Parkinson's and Huntington's can disrupt it. People with Parkinson's disease, for example, tend to have increased saccadic latencies. Consequently, the quantitative measurement of saccadic eye movements has received considerable attention as a potential biomarker for neurodegenerative conditions. A lot more can be learned about the brain in both health and disease by observing what happens to eye movements when the function of specific brain areas is perturbed. Deep brain stimulation is a surgical intervention used for the management of a range of neurological conditions including Parkinson's disease, in which stimulating electrodes are placed in specific brain areas including several sites in the basal ganglia. Eye movement measurements can then be made with the stimulator systems both off and on and the results compared. With suitable experimental design, this approach can be used to study the pathophysiology of the disease being treated, the mechanism by which DBS exerts it beneficial effects, and even aspects of normal neurophysiology.

This paper describes the use of quantitative measurement of eye movements in conjunction with stimulation of focal areas of the deep brain in order to study physiology, pathophysiology, and the mechanisms of deep brain stimulation.

使用Saccadometry与深部脑刺激来研究正常和病理脑功能

The oculomotor system involves a large number of brain areas including parts of the basal ganglia, and various neurodegenerative diseases including ...

Imaging Metals in Brain Tissue by Laser Ablation - Inductively Coupled Plasma - Mass Spectrometry (LA-ICP-MS)

Metals are found ubiquitously throughout an organism, with their biological role dictated by both their chemical reactivity and abundance within a specific anatomical region. Within the brain, metals have a highly compartmentalized distribution, depending on the primary function they play within the central nervous system. Imaging the spatial distribution of metals has provided unique insight into the biochemical architecture of the brain, allowing direct correlation between neuroanatomical regions and their known function with regard to metal-dependent processes. In addition, several age-related neurological disorders feature disrupted metal homeostasis, which is often confined to small regions of the brain that are otherwise difficult to analyze. Here, we describe a comprehensive method for quantitatively imaging metals in the mouse brain, using laser ablation - inductively coupled plasma - mass spectrometry (LA-ICP-MS) and specially designed image processing software. Focusing on iron, copper and zinc, which are three of the most abundant and disease-relevant metals within the brain, we describe the essential steps in sample preparation, analysis, quantitative measurements and image processing to produce maps of metal distribution within the low micrometer resolution range. This technique, applicable to any cut tissue section, is capable of demonstrating the highly variable distribution of metals within an organ or system, and can be used to identify changes in metal homeostasis and absolute levels within fine anatomical structures.

Imaging Metals in Brain Tissue by Laser Ablation - Inductively Coupled Plasma - Mass Spectrometry LA-ICP-MS

Quantitatively mapping metals in tissue by laser ablation - inductively coupled plasma - mass spectrometry (LA-ICP-MS) is a sensitive analytical technique that can provide new insight into how metals participate in normal function and disease processes. Here, we describe a protocol for quantitatively imaging metals in thin sections of mouse neurological tissue.

在脑组织成像金属激光剥蚀 - 电感耦合等离子体 - 质谱法（LA-ICP-MS）

Metals are found ubiquitously throughout an organism, with their biological role dictated by both their chemical reactivity and abundance within a ...

Absolute Quantification of A&#946;1-42 in CSF Using a Mass Spectrometric Reference Measurement Procedure

Alzheimer's disease (AD) is the most common neurodegenerative disease among the elderly and accounts for 60-80% of all cases of dementia. Currently, the diagnosis of AD is based on cognitive tests and mental state exams, but the peptide amyloid-beta (A&#946;) in cerebrospinal fluid (CSF) is increasingly used in clinical trials and settings. As for most protein and peptide biomarkers, quantification is performed using antibody-based techniques, such as enzyme-linked immunosorbent assay (ELISA). However, intra- and inter-laboratory variability in these assays hamper its use as a diagnostic marker in clinical routine.
An antibody-independent Reference Measurement Procedure (RMP) was developed based on solid-phase extraction (SPE) and liquid chromatography (LC)-tandem mass spectrometry (MS/MS), where stable, isotope-labeled A&#946; peptides were used as internal standards, enabling absolute quantification. A high-resolution quadrupole-orbitrap hybrid instrument was used for the measurements. The method allows for the quantification of CSF A&#946;1-42 between 150-4,000 pg/mL.

A reference measurement procedure for the absolute quantification of A&#946;1-42 in human CSF based on solid-phase extraction and liquid chromatography tandem mass spectrometry is described.

Aβ的绝对定量<sub&gt; 1-42</sub&gt;在CSF使用质谱参考测量程序

Alzheimer's disease (AD) is the most common neurodegenerative disease among the elderly and accounts for 60-80% of all cases of dementia. Currently, the ...

Repeated Measurement of Respiratory Muscle Activity and Ventilation in Mouse Models of Neuromuscular Disease

Accessory respiratory muscles help to maintain ventilation when diaphragm function is impaired. The following protocol describes a method for repeated measurements over weeks or months of accessory respiratory muscle activity while simultaneously measuring ventilation in a non-anesthetized, freely behaving mouse. The technique includes the surgical implantation of a radio transmitter and the insertion of electrode leads into the scalene and trapezius muscles to measure the electromyogram activity of these inspiratory muscles. Ventilation is measured by whole-body plethysmography, and animal movement is assessed by video and is synchronized with electromyogram activity. Measurements of muscle activity and ventilation in a mouse model of amyotrophic lateral sclerosis are presented to show how this tool can be used to investigate how respiratory muscle activity changes over time and to assess the impact of muscle activity on ventilation. The described methods can easily be adapted to measure the activity of other muscles or to assess accessory respiratory muscle activity in additional mouse models of disease or injury.

This paper introduces a method for repeated measurements of ventilation and respiratory muscle activity in a freely behaving amyotrophic lateral sclerosis (ALS) mouse model throughout disease progression with whole-body plethysmography and electromyography via an implanted telemetry device.

在神经肌肉疾病的模型小鼠呼吸肌肉活动和通风的重复测量

Accessory respiratory muscles help to maintain ventilation when diaphragm function is impaired. The following protocol describes a method for repeated ...