作者要感谢他们的合作者在研究采用的内存眼球运动监测，特别感谢Eyal莱因戈尔德，沉基业，尼尔J.科恩，罗伯特R. Althoff，德博拉Hannula，和Dave沃伦。这项工作已经由自然科学和工程研究理事会（NSERC），加拿大卫生研究所（CIHR）加拿大研究所，加拿大研究教席计划和创新（CFI）加拿大基金会的支持。

在数据采集过程中使用的设备眼动仪在目前的协议使用眼动仪是一个EyeLink II系统（SR研究有限公司，加拿大安大略省Mississauga）。这头安装，基于视频的眼动仪记录眼位在X，Y坐标在500或250赫兹的采样率框架的空间分辨率&lt;0.1 ​​°，。一台摄像机用于监视发送红外标记与会者观看显示器的四个角落放置传感器头部的位置。两个额外的摄像机被安装在位于眼睛下面的头带，要注意瞳孔和角膜反射的红外照明。可根据眼位，瞳孔和角膜反射，或只瞳孔。眼动仪的软垫的头带可调节两架飞机的舒适适合成年参与者的头部大小。眼动仪可容纳大多数的眼镜和隐形眼镜。 计算机两台电脑是用来支持眼球运动记录。一台计算机作为显示计算机，其中介绍的校准屏幕，必要的任务说明和实验范式中使用的图像向与会者。显示计算机的详细信息数据收集第二，电脑主机，然后由管辖的参数。电脑主机，计算实时的目光位置和记录眼球运动以后的分析数据，以及任何按下按钮或键盘响应的参与者。参与者的眼动仪的安装和操作都是通过PC主机。 软件在这个协议中，通过实验刺激的时机和顺序提交给与会者，和眼睛的位置是由PC主机收集的方式，通过实验生成器，专门SR研究有限公司开发的一个软件程序，编程眼动仪主机电脑接口。然而，刺激呈现也可以通过其他软件程序（例如，简报，神经行为系统;奥尔巴尼，CA）进行。眼球运动数据，以一系列固定和扫视事件的转换时间锁定为刺激呈现（或其他外部事件）是通过主机和SR研究有限公司开发的一个软件程序，数据查看器，可与审问;然而，再次，其他程序可以用来获得所需的眼球运动措施。在这里，录制品和扫视检测依赖于一个网络分析器，分成有意义的状态（扫视，闪烁和录制品）的原料眼球运动样本。如果连续两个眼球运动样本的速度超过每秒22度，距离超过0.1 °，样品标示为扫视。如果学生是失踪3个或更多的样品，眼球活动标记为一个数据流内闪烁。非扫视和非闪烁的活动被认为是复制品。 眼球跟踪程序。 下面，我们详细的程序，获得每个参与者的眼球运动记录。 <ol><li>同意。参与者设置之前，参与者眼动仪的头带。实验者解释，头部和眼睛的位置是通过头带摄像机监视的参与者。 </li><li>对于一个给定的实验范式，与会者都坐在一个固定的距离从监视器参与者之间保持相同的视觉角度。 </li><li>眼动仪的相机设置。眼动仪头盔头盔的调整，使舒适，不会移动，但周围的头部不舒服。头盔是进一步调整，使头摄像头，可以发送红外线照射到显示屏上的外部标记。坐落在个人棒，从头盔，允许所有轴调整延长每眼相机。眼相机是定位略低于稍微远离每只眼睛不妨碍参与者S的显示器（见图1）。该相机的重点，得到一个明确的瞳孔和角膜反射和稳定的图像。在主机上的状态面板显示瞳孔和角膜反射是否正在收购。照明阈值可以进行调整，以获得最稳定的瞳孔和角膜反射录音。实验者，然后选择适当的实验范式，并指定为随后的眼球运动录音的文件名。 <img src="/files/ftp_upload/2108/2108fig1.jpg" alt="图1" /> 图1。基于视频头式眼动仪（左）和显示器和PC主机（右）的例子。请注意，在参与者面前的显示器，显示瞳孔的位置（蓝十字点）和角膜反射（十字黄点），以帮助在相机设置。 </li><li>校准。为了获得准确的显示屏上的眼睛位置录音，校准程序启动以下的相机设置。校准过程中，学员们指示看看了一系列的目标出现在显示屏上的不同地点。通常情况下，9个目标地点，但可以用尽可能少的三个目标地点进行校准。从这9个记录地点，眼睛在屏幕上的任何点的位置可插。 </li><li>验证。以下校准，验证程序是用于检查眼球运动记录的准确性。相同的9个目标地点，提供给参与者注视，所不同的是在当前的固定位置和先前记录的固定位置计算。如果记录的位置之一（或两者）的眼睛平均精度超过了可接受的水平（平均误差小于0.5 °，最大的任何一点的误差&lt;1.0 °），校准和验证程序是重复的。实验者可以接受的精度水平，确定是否应单眼或双眼眼球运动记录。在单眼记录的情况下，实验者可以决定是要记录眼数据，或PC主机可以自动选择较低的平均误差和最大误差较低的眼睛。虽然校准和验证可能会出现轻微的头部运动，它是有与会者坐在仍然尽可能有利的。参与者也没有预见到即将到来的目标位置的指示，但而移动，并注视故意，目标只有当他们出现，停留在迷恋的目标，直至消失。 </li><li>实验范式。提交的参与者（例如，“自由请查看下面的图像。的”），特别是一个给定的范式的指令和数据记录发起会话。 </li><li>漂移校正。之前的每一个实验的刺激开始，漂移校正，可以让参与者在中央固定目标，同时按下手柄或键盘上的一个按钮来接受固定的位置。当采用漂移校正，审判将不会开始，直到成功的参与者在中央固定区域内的中心目标2.0 ° fixates。如果参与者始终fixates大于2.0 °，参与者必须再次进行校准和验证程序。任何差别小于2.0 °之间的记录眼睛的位置和在校准/验证记录，获得初始的固定位置，然后注意到的错误，并在数据。 </li><li>刺激呈现。眼位，包括固定和扫视事件，记录了每个刺激呈现以下漂移校正程序。实验者可以通过电脑主机的检查在线监控录音的准确性。 </li><li>以下的眼球运动记录会话，眼球运动特点的措施，为不同类型的图像（如小说，反复）扫描模式的各种软件包，如SR研究公司的数据查看器，可以推导出。此外，观看刺激（如保持不变，操纵）的特定区域，可以通过分析眼球运动方面的特点实验者绘制感兴趣的地区，无论是在范式编程阶段或数据收集后，为每个图像创建。 </li><li>与会者在会议结束实验的目的就盘问。 </li></ol>代表性的成果从眼球运动的录音，包括描述整体观看到的图像（包括每个固定/扫视的特点）的措施，以及措施，描述了观看的模式，已指示利益的特定区域内，可以得出多种措施图像3。整体观看图像的措施可能包括（但不仅限于）：录制品的数量和图像扫视，平均每个固定时间，和总金额的观赏时间，花了行了吧上图像。描述查看特定地区的利益格局的措施可能包括（但不限于）：感兴趣的区域的录制品的数量，一个感兴趣的区域内度过的时间，和转换次数进/出一个感兴趣的区域。此外，这些措施可能源于眼球运动的录音，轮廓（即，何时机瓦特初）一个特定的眼球运动事件发生，如时，刺激呈现后，眼睛上的利益的特定区域，当第一扫视图像注视，熵（约束/随机性）在固有眼球运动模式的序列。 对于任何给定的图像，眼球运动的措施可以被迷恋的眼睛，当多久。要获得内存的索引，我们可以给观众不同的类型或接触到不同的图像集的的金额，然后比较在这些集和跨与会者观看模式。例如，重复图像探测内存，扫描模式，可以对比已被视为整个测试会话多次新颖的图像和图像。观看图像多次在减少整体观看图像2，4-5，8测试在整个会议的结果。这可以看出，在图2。在此代表结果11里格斯等，与会者认为面临的小说图片曾在五个测试块;。日益暴露，有一个观众的面孔的录制品的数量有所减少。为了探测图像的具体细节内存，可以扫描模式已多次在其原始的，不变的形式（重复图像），并反复测试会议，同样被视为整个图像浏览图像之间的对比，但已改变在现场的一些元素在最后曝光（操纵图像）。在这种情况下，扫描模式的差异是吸引相同的，重复的图像2-3，7-8不变地区相比已在操纵的形象改变的区域。然而，如眼球运动的内存指数在某些人群，如健康的老年人和失忆由于内侧颞叶损伤的患者时评估他们的记忆场景中的对象之间的空间关系，目前，在图3概述2，8。因此，从眼球运动监测的结果可用于参与者群体之间的对比记忆体具有不同的神经心理状态2-3， 8-10。 <img src="/files/ftp_upload/2108/2108fig2.jpg" alt="图2" /> 图2。眼球运动揭示了重复内存在这种代表性的结果，从我们的实验室[11]，与会者观看面临在研究5块;数量增加（1-5）收看，录制品的数量的面孔下降。 <img src="/files/ftp_upload/2108/2108fig3.jpg" alt="图3" /> 图3。眼球运动揭示改变细节的记忆，年轻的成年人[2（实验1，2），8（免费观看条件）]的更大比例的总眼录制品在操纵的形象，已发生了变化从一个关键区域前观看相比，当该地区没有发生变化，作为在新的和重复图像。内存等的影响，没有健康的老年人[8（免费观看，并在健忘症患者的状况[2]。

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	<li>Bradfield, A. L., Wells, G. L., Olsen, E. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+damaging+effect+of+confirming+feedback+on+the+relation+between+eyewitness+certainty+and+identification+accuracy.">The damaging effect of confirming feedback on the relation between eyewitness certainty and identification accuracy.</a> J. Appl. Psychol. 87 (1), 112-120 (2002).</li><li>Ryan, J. D., Althoff, R. R., Whitlow, S., Cohen, N. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Amnesia+is+a+deficit+in+relational+memory.">Amnesia is a deficit in relational memory.</a> Psych. Sci. 11 (6), 454-461 (2000).</li><li>Ryan, J. D., Cohen, N. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+nature+of+change+detection+and+online+representations+of+scenes.">The nature of change detection and online representations of scenes.</a> J. Exp. Psych. Hum. Percept. Perfor. 30, 98-1015 (2004).</li><li>Althoff, R. R., Cohen, N. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Eye-movement-based+memory+effect:+a+re-processing+effect+in+face+perception.">Eye-movement-based memory effect: a re-processing effect in face perception.</a> J. Exp. Psychol. Learn Mem. Cog. 25 (4), 997-1010 (1999).</li><li>Heisz, J. J., Shore, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=More+efficient+scanning+for+familiar+faces.">More efficient scanning for familiar faces.</a> J. Vis. 8 (1), 1-10 (2008).</li><li>Parker, R. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Picture+processing+during+recognition.">Picture processing during recognition.</a> J. Exp. Psych. Hum. Percept. Perfor. 4, 284-293 (1978).</li><li>Hollingworth, A., Williams, C. C., Henderson, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=To+see+and+remember:+Visually+specific+information+is+retained+in+memory+from+previously+attended+objects+in+natural+scenes.">To see and remember: Visually specific information is retained in memory from previously attended objects in natural scenes.</a> Psychon Bull. Rev. 8, 761-768 (2001).</li><li>Ryan, J. D., Leung, G., Turk-Browne, N. B., Hasher, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+age-related+changes+in+inhibition+and+binding+using+eye+movement+monitoring.">Assessment of age-related changes in inhibition and binding using eye movement monitoring.</a> Psychol. Aging. 22 (2), 239-250 (2007).</li><li>Hannula, D. E., Ryan, J. D., Tranel, D., Cohen, N. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rapid+onset+relational+memory+effects+are+evident+in+eye+movement+behavior,+but+not+in+hippocampal+amnesia.">Rapid onset relational memory effects are evident in eye movement behavior, but not in hippocampal amnesia.</a> J. Cog. Neuro. 19 (10), 1690-1705 (2007).</li><li>Bate, S., Haslam, C., Tree, J. J., Hodgson, T. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evidence+of+an+eye+movement-based+memory+effect+in+congenital+prosopsagnosia.">Evidence of an eye movement-based memory effect in congenital prosopsagnosia.</a> Cortex. 44 (7), 806-819 (2008).</li><li>Riggs, L., McQuiggan, D., Chan, J., Anderson, A. K., Ryan, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Emotion-+modulated+viewing+of+faces+by+association.">Emotion- modulated viewing of faces by association.</a> , .</li><li>Kramer, A. F., McCarley, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Oculomotor+behaviour+as+a+reflection+of+attention+and+memory+processes:+Neural+mechanisms+and+applications+to+human+factors.">Oculomotor behaviour as a reflection of attention and memory processes: Neural mechanisms and applications to human factors.</a> Theor. Issues Ergon. Sci. 4 (1), 21-55 (2003).</li></ol>

眼球运动监测是一种高效，有用的工具，用以评估记忆功能，在各种不同的人群。本协议描述了一个头戴式视频眼动仪的使用，但该协议可以很容易地适应远程眼球跟踪设备的使用，远程眼球跟踪取下头盔调整的需要，简化了相机调整。然而，与远程的眼动仪，头部运动必须受到限制，以保持眼睛录音的准确性。眼球运动的精确校准是最重要的获取有用的，并解释，数据。 ?...

A correction was made to <a href="http://www.jove.com/index/details.stp?id=2108">Eye Movement Monitoring of Memory</a>. There was an error in the author, Douglas A. McQuiggan's, name. The author's name has been corrected to:
Douglas A. McQuiggan
instead of:
Doug McQuiggan

A correction to an author's name has been made for the article Eye Movement Monitoring of Memory.

Erratum: Eye Movement Monitoring of Memory

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<td>Eye Tracker</td>
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eye movement monitoring of memory

明确的（通常是口头的）报告通常用于研究记忆体（如“告诉我你还记得有关的人，你看到在昨天的银行。”），然而，这类报告常常可以不可靠的或敏感的反应偏差<sup&gt; 1</sup&gt;，并可能在一些参与者的人口用不上。此外，明确报告只透露信息时已达意识，不能发表评论时的记忆是在加工过程中访问，不管信息是否有意识地在随后访问。眼球运动监测（眼球跟踪）提供了一个工具，其中内存可以不要求参加者在他们的记忆的内容发表评论探测，并访问这些记忆可以揭示上线<sup&gt; 2,3</sup&gt;。基于视频的眼球追踪器（无论是头部安装或远程）使用的相机和红外标记系统，以研究在每只眼睛的瞳孔和角膜反射，作为参与者看法显示器。对于头戴式眼跟踪，红外标记也可用于确定头部位置，以便头部运动和更精确的眼位的本地化。在这里，我们展示了一个头戴式眼跟踪系统的使用调查中的神经功能完好和神经功能受损的成年人的内存性能。眼动监控程序开始与参与者的位置，眼动仪，头部和眼睛的摄像头设置。校准和验证的程序进行，以确保眼睛的位置记录的准确性。实时录音，X，Y坐标显示屏上的位置，然后将其转换，用来描述的时间段中，映入眼帘的是静态的（即录制品）在运动与（即扫视）。录制品和扫视发病时间锁定/偏移一个可视化显示或其他外部事件（如按下按钮）。实验操作的构造，研究如何以及何时通过以往的经验不同类型的录制品和扫视模式改变。扫描模式，以新的图像从扫描模式，图像先前已研究不同的程度上揭示了记忆的影响<sup&gt; 2，4-5</sup&gt;。存储器也可讯问其特殊性，例如，眼球运动模式之间的相同和以前学过的图像改变版本不同显示存储在内存中的更改细节<sup&gt; 2-3，6-8</sup&gt;。内存这些指标可以比较整个参与者的人群，从而提供了有力的工具，其中研究组织在健康个体的记忆，以及发生与神经侮辱或下降到内存中的具体变化<sup&gt; 2-3，8-10</sup&gt;。

Watch this Scientific Journal Video about Eye Movement Monitoring of Memory at JoVE.com

Eye Movement Monitoring of Memory

眼球运动监测（或眼睛跟踪），揭示了在太空中的眼睛流连忘返，当多久。在这里，我们展示了如何可用于眼跟踪调查内存在多个参与者群体的完整性，而不需要口头或以其他方式明确，报告。

眼运动监测内存

Explicit (often verbal) reports are typically used to investigate memory (e.g. "Tell me what you remember about the person you saw at the bank ...

Research

JoVE Journal

Neuroscience

14.5K 次观看.  Rotman Research Institute. 眼球运动监测（或眼睛跟踪），揭示了在太空中的眼睛流连忘返，当多久。在这里，我们展示了如何可用于眼跟踪调查内存在多个参与者群体的完整性，而不需要口头或以其他方式明确，报告。

视频: Eye Movement Monitoring of Memory

Experimental Models for Study of Retinal Pigment Epithelial Physiology and Pathophysiology

We have developed a cell culture procedure that can produce large quantities of confluent monolayers of primary human fetal retinal pigment epithelium (hfRPE) cultures with morphological, physiological and genetic characteristics of native human RPE. These hfRPE cell cultures exhibit heavy pigmentation, and electron microscopy show extensive apical membrane microvilli. The junctional complexes were identified with immunofluorescence labeling of various tight junction proteins. Epithelial polarity and function of these easily reproducible primary cultures closely resemble previously studied mammalian models of native RPE, including human. These results were extended by the development of therapeutic interventions in several animal models of human eye disease. We have focused on strategies for the removal of abnormal fluid accumulation in the retina or subretinal space. The extracellular subretinal space separates the photoreceptor outer segments and the apical membrane of the RPE and is critical for maintenance of retinal attachments and a whole host of RPE/retina interactions.

We provide a reproducible method for culturing confluent monolayers of human fetal retinal pigment epithelial cells (hfRPE) cells that exhibit morphology, physiology, polarity, and protein and gene expression patterns of adult native tissue. This work has been extended to an animal model of several eye diseases.

视网膜色素上皮的生理与病理生理学研究的实验模型

We have developed a cell culture procedure that can produce large quantities of confluent monolayers of primary human fetal retinal pigment epithelium ...

科研

神经科学

Physiological, Morphological and Neurochemical Characterization of Neurons Modulated by Movement

The role of individual neurons and their function in neuronal circuits is fundamental to understanding the neuronal mechanisms of sensory and motor functions. Most investigations of sensorimotor mechanisms rely on either examination of neurons while an animal is static1,2 or record extracellular neuronal activity during a movement.3,4 While these studies have provided the fundamental background for sensorimotor function, they either do not evaluate functional information which occurs during a movement or are limited in their ability to fully characterize the anatomy, physiology and neurochemical phenotype of the neuron. A technique is shown here which allows extensive characterization of individual neurons during an in vivo movement. This technique can be used not only to study primary afferent neurons but also to characterize motoneurons and sensorimotor interneurons. Initially the response of a single neuron is recorded using electrophysiological methods during various movements of the mandible followed by determination of the receptive field for the neuron. A neuronal tracer is then intracellularly injected into the neuron and the brain is processed so that the neuron can be visualized with light, electron or confocal microscopy (Fig. 1). The detailed morphology of the characterized neuron is then reconstructed so that neuronal morphology can be correlated with the physiological response of the neuron (Figs. 2,3). In this communication important key details and tips for successful implementation of this technique are provided. Valuable additional information can be determined for the neuron under study by combining this method with other techniques. Retrograde neuronal labeling can be used to determine neurons with which the labeled neuron synapses; thus allowing detailed determination of neuronal circuitry. Immunocytochemistry can be combined with this method to examine neurotransmitters within the labeled neuron and to determine the chemical phenotypes of neurons with which the labeled neuron synapses. The labeled neuron can also be processed for electron microscopy to determine the ultrastructural features and microcircuitry of the labeled neuron. Overall this technique is a powerful method to thoroughly characterize neurons during in vivo movement thus allowing substantial insight into the role of the neuron in sensorimotor function.

A technique is described to quantify the in vivo physiological response of mammalian neurons during movement and correlate the physiology of the neuron with neuronal morphology, neurochemical phenotype and synaptic microcircuitry.

由运动神经元调制生理，形态学和神经化学的表征

The role of individual neurons and their function in neuronal circuits is fundamental to understanding the neuronal mechanisms of sensory and motor ...

Coherence between Brain Cortical Function and Neurocognitive Performance during Changed Gravity Conditions

Previous studies of cognitive, mental and/or motor processes during short-, medium- and long-term weightlessness have only been descriptive in nature, and focused on psychological aspects. Until now, objective observation of neurophysiological parameters has not been carried out - undoubtedly because the technical and methodological means have not been available -, investigations into the neurophysiological effects of weightlessness are in their infancy (Schneider et al. 2008).
While imaging techniques such as positron emission tomography (PET) and magnetic resonance imaging (MRI) would be hardly applicable in space, the non-invasive near-infrared spectroscopy (NIRS) technique represents a method of mapping hemodynamic processes in the brain in real time that is both relatively inexpensive and that can be employed even under extreme conditions. The combination with electroencephalography (EEG) opens up the possibility of following the electrocortical processes under changing gravity conditions with a finer temporal resolution as well as with deeper localization, for instance with electrotomography (LORETA).
Previous studies showed an increase of beta frequency activity under normal gravity conditions and a decrease under weightlessness conditions during a parabolic flight (Schneider et al. 2008a+b). Tilt studies revealed different changes in brain function, which let suggest, that changes in parabolic flight might reflect emotional processes rather than hemodynamic changes. However, it is still unclear whether these are effects of changed gravity or hemodynamic changes within the brain. Combining EEG/LORETA and NIRS should for the first time make it possible to map the effect of weightlessness and reduced gravity on both hemodynamic and electrophysiological processes in the brain. Initially, this is to be done as part of a feasibility study during a parabolic flight. Afterwards, it is also planned to use both techniques during medium- and long-term space flight.
It can be assumed that the long-term redistribution of the blood volume and the associated increase in the supply of oxygen to the brain will lead to changes in the central nervous system that are also responsible for anaemic processes, and which can in turn reduce performance (De Santo et al. 2005), which means that they could be crucial for the success and safety of a mission (Genik et al. 2005, Ellis 2000).
Depending on these results, it will be necessary to develop and employ extensive countermeasures. Initial results for the MARS500 study suggest that, in addition to their significance in the context of the cardiovascular and locomotor systems, sport and physical activity can play a part in improving neurocognitive parameters. Before this can be fully established, however, it seems necessary to learn more about the influence of changing gravity conditions on neurophysiological processes and associated neurocognitive impairment.

The effect of weightlessness and hypergravity on both hemodynamic and electrophysiological processes in the brain is going to be followed during parabolic flight by EEG and NIRS techniques. 
A feasibility study of a more complex experiment, which is planned to carry out during medium- and long-term space flight.

大脑皮质功能和神经在更改重力条件下的性能之间的一致性

Previous studies of cognitive, mental and/or motor processes during short-, medium- and long-term weightlessness have only been descriptive in nature, and ...

Using Affordable LED Arrays for Photo-Stimulation of Neurons

Standard slice electrophysiology has allowed researchers to probe individual components of neural circuitry by recording electrical responses of single cells in response to electrical or pharmacological manipulations1,2. With the invention of methods to optically control genetically targeted neurons (optogenetics), researchers now have an unprecedented level of control over specific groups of neurons in the standard slice preparation. In particular, photosensitive channelrhodopsin-2 (ChR2) allows researchers to activate neurons with light3,4. By combining careful calibration of LED-based photostimulation of ChR2 with standard slice electrophysiology, we are able to probe with greater detail the role of adult-born interneurons in the olfactory bulb, the first central relay of the olfactory system. Using viral expression of ChR2-YFP specifically in adult-born neurons, we can selectively control young adult-born neurons in a milieu of older and mature neurons. Our optical control uses a simple and inexpensive LED system, and we show how this system can be calibrated to understand how much light is needed to evoke spiking activity in single neurons. Hence, brief flashes of blue light can remotely control the firing pattern of ChR2-transduced newborn cells.

Adult-born neurons expressing ChR2 can be manipulated in slice electrophysiological preparations in order to examine their contribution towards the function of olfactory neural circuits.

使用光刺激神经元的负担得起的LED阵列

Standard slice electrophysiology has allowed researchers to probe individual components of neural circuitry by recording electrical responses of single ...

Selective Viral Transduction of Adult-born Olfactory Neurons for Chronic in vivo Optogenetic Stimulation

Local interneurons are continuously regenerated in the olfactory bulb of adult rodents1-3. In this process, called adult neurogenesis, neural stem cells in the walls of the lateral ventricle give rise to neuroblasts that migrate for several millimeters along the rostral migratory stream (RMS) to reach and incorporate into the olfactory bulb. To study the different steps and the impact of adult-born neuron integration into preexisting olfactory circuits, it is necessary to selectively label and manipulate the activity of this specific population of neurons. The recent development of optogenetic technologies offers the opportunity to use light to precisely activate this specific cohort of neurons without affecting surrounding neurons4,5. Here, we present a series of procedures to virally express Channelrhodopsin2(ChR2)-YFP in a temporally restricted cohort of neuroblasts in the RMS before they reach the olfactory bulb and become adult-born neurons. In addition, we show how to implant and calibrate a miniature LED for chronic in vivo stimulation of ChR2-expressing neurons.

Selective Viral Transduction of Adult-born Olfactory Neurons for Chronic in vivo Optogenetic Stimulation

Adult-born neurons of the olfactory bulb can be optogenetically controlled using Channelrhodopsin2-expressing lentiviral injection in the rostral migratory stream and chronic photostimulation with an implanted miniature LED.

慢性成人出生的嗅觉神经元的选择性病毒转导在体内 Optogenetic刺激

Local interneurons are continuously regenerated in the olfactory bulb of adult rodents1-3. In this process, called adult neurogenesis, neural stem cells ...

Rapid Determination of the Thermal Nociceptive Threshold in Diabetic Rats

Painful diabetic neuropathy (PDN) is characterized by hyperalgesia i.e., increased sensitivity to noxious stimulus, and allodynia i.e., hypersensitivity to normally innocuous stimuli1. Hyperalgesia and allodynia have been studied in many different rodent models of diabetes mellitus2. However, as stated by B&ouml;lcskei et al, determination of "pain" in animal models is challenging due to its subjective nature3. Moreover, the traditional methods used to determine behavioral responses to noxious thermal stimuli usually lack reproducibility and pharmacological sensitivity3. For instance, by using the hot-plate method of Ankier4, flinch, withdrawal and/or licking of either hind- and/or fore-paws is quantified as reflex latencies at constant high thermal stimuli (52-55 &deg;C). However, animals that are hyperalgesic to thermal stimulus do not reproducibly show differences in reflex latencies using those supra-threshold temperatures3,5. As the recently described method of B&ouml;lcskei et al.6, the procedures described here allows for the rapid, sensitive and reproducible determination of thermal nociceptive thresholds (TNTs) in mice and rats. The method uses slowly increasing thermal stimulus applied mostly to the skin of mouse/rat plantar surface. The method is particularly sensitive to study anti-nociception during hyperalgesic states such as PDN. The procedures described bellow are based on the ones published in detail by Alm&aacute;si et al 5 and B&ouml;lcskei et al 3. The procedures described here have been approved the Laboratory Animal Care and Use Committee (LACUC), Wright State University.

Here, we describe a rapid reliable and simple procedure to determine the lowest temperature at which rats or mice show nocifensive behavior, i.e. the thermal nociceptive threshold (TNT). This method applies a slowly increasing thermal stimulus allowing precise and reproducible estimation of TNTs with minimum, if any, stress to the animals.

糖尿病大鼠的热痛觉阈值的快速测定

Painful diabetic neuropathy (PDN) is characterized by hyperalgesia i.e., increased sensitivity to noxious stimulus, and allodynia i.e., hypersensitivity ...

Optical Recording of Suprathreshold Neural Activity with Single-cell and Single-spike Resolution

Signaling of information in the vertebrate central nervous system is often carried by populations of neurons rather than individual neurons. Also propagation of suprathreshold spiking activity involves populations of neurons. Empirical studies addressing cortical function directly thus require recordings from populations of neurons with high resolution. Here we describe an optical method and a deconvolution algorithm to record neural activity from up to 100 neurons with single-cell and single-spike resolution. This method relies on detection of the transient increases in intracellular somatic calcium concentration associated with suprathreshold electrical spikes (action potentials) in cortical neurons. High temporal resolution of the optical recordings is achieved by a fast random-access scanning technique using acousto-optical deflectors (AODs)1. Two-photon excitation of the calcium-sensitive dye results in high spatial resolution in opaque brain tissue2. Reconstruction of spikes from the fluorescence calcium recordings is achieved by a maximum-likelihood method. Simultaneous electrophysiological and optical recordings indicate that our method reliably detects spikes (&gt;97% spike detection efficiency), has a low rate of false positive spike detection (&lt; 0.003 spikes/sec), and a high temporal precision (about 3 msec) 3. This optical method of spike detection can be used to record neural activity in vitro and in anesthetized animals in vivo3,4.

Understanding the function of the vertebrate central nervous system requires recordings from many neurons because cortical function arises on the level of populations of neurons. Here we describe an optical method to record suprathreshold neural activity with single-cell and single-spike resolution, dithered random-access scanning. This method records somatic fluorescence calcium signals from up to 100 neurons with high temporal resolution. A maximum-likelihood algorithm deconvolves the underlying suprathreshold neural activity from the somatic fluorescence calcium signals. This method reliably detects spikes with high detection efficiency and a low rate of false positives and can be used to study neural populations in vitro and in vivo.

阈上的神经活动的单细胞和单穗的分辨率的光学的方法记录

Signaling of information in the vertebrate central nervous system is often carried by populations of neurons rather than individual neurons. Also ...

Observation of the Ciliary Movement of Choroid Plexus Epithelial Cells Ex Vivo

The choroid plexus is located in the ventricular wall of the brain, the main function of which is believed to be production of cerebrospinal fluid. Choroid plexus epithelial cells (CPECs) covering the surface of choroid plexus tissue harbor multiple unique cilia, but most of the functions of these cilia remain to be investigated. To uncover the function of CPEC cilia with particular reference to their motility, an ex vivo observation system was developed to monitor ciliary motility during embryonic, perinatal and postnatal periods. The choroid plexus was dissected out of the brain ventricle and observed under a video-enhanced contrast microscope equipped with differential interference contrast optics. Under this condition, a simple and quantitative method was developed to analyze the motile profiles of CPEC cilia for several hours ex vivo. Next, the morphological changes of cilia during development were observed by scanning electron microscopy to elucidate the relationship between the morphological maturity of cilia and motility. Interestingly, this method could delineate changes in the number and length of cilia, which peaked at postnatal day (P) 2, while the beating frequency reached a maximum at P10, followed by abrupt cessation at P14. These techniques will enable elucidation of the functions of cilia in various tissues. While related techniques have been published in a previous report1, the current study focuses on detailed techniques to observe the motility and morphology of CPEC cilia ex vivo.

Observation of the Ciliary Movement of Choroid Plexus Epithelial Cells Ex Vivo

In this study, a detailed light microscopic technique was optimized for real-time observation and analysis of the motion of CPEC cilia ex vivo together with an electron microscopic method for ultrastructural analysis.

脉络丛上皮细胞的纤毛运动的观察<em&gt;离体</em

The choroid plexus is located in the ventricular wall of the brain, the main function of which is believed to be production of cerebrospinal fluid. ...

Characterizing the Relationship Between Eye Movement Parameters and Cognitive Functions in Non-demented Parkinson's Disease Patients with Eye Tracking

Cognitive impairment is a common phenomenon in Parkinson&#8217;s disease that has implications on the prognosis. A simple, noninvasive and objective proxy measurement of cognitive function in Parkinson&#8217;s disease will be helpful in detecting early cognitive decline. As a physiological metric, eye movement parameter is not confounded by the subject&#39;s attributes and intelligence and can function as a proxy marker if it correlates with cognitive functions. To this end, this study explored the relationship between the eye movement parameters and performance in cognitive tests in multiple domains. In the experiment, a visual search task with eye tracking was set up, where subjects were asked to look for a number embedded in an array of alphabets scattered randomly on a computer screen. The differentiation between the number and the alphabet is an overlearned task such that the confounding effect of cognitive ability on the eye movement parameters is minimized. The average saccadic amplitude and fixation duration were captured and calculated during the visual search task. The cognitive assessment battery covered domains of frontal-executive functions, attention, verbal and visual memory. It was found that prolonged fixation duration was associated with poorer performance in verbal fluency, visual and verbal memory, allowing further exploration on the use of eye movement parameters as proxy markers for cognitive function in Parkinson&#8217;s disease patients. The experimental paradigm has been found to be highly tolerable in our group of Parkinson&#39;s disease patients and could be applied transdiagnostically to other disease entities for similar research questions.

Here, we present a protocol to study the relationship between the eye movement parameters and cognitive functions in non-demented Parkinson&#39;s disease patients. The experiment used an eye tracker to measure the saccadic amplitude and fixation duration in a visual search task. The correlation with performance in multi-domain cognitive tasks was subsequently measured.

描述眼动参数与无视力帕金森病患者眼动功能的关系

Cognitive impairment is a common phenomenon in Parkinson&#8217;s disease that has implications on the prognosis. A simple, noninvasive and objective proxy ...

Immunostaining of Whole-Mount Retinas with the CLARITY Tissue Clearing Method

The tissue hydrogel delipidation method (CLARITY), originally developed by the Deisseroth laboratory, has been modified and widely used for immunostaining and imaging of thick brain samples. However, this advanced technology has not yet been used for whole-mount retinas. Although the retina is partially transparent, its thickness of approximately 200 &#181;m (in mice) still limits the penetration of antibodies into the deep tissue as well as reducing light penetration for high-resolution imaging. Here, we adapted the CLARITY method for whole-mount mouse retinas by polymerizing them with an acrylamide monomer to form a nanoporous hydrogel and then clearing them in sodium dodecyl sulfate to minimize protein loss and avoid tissue damage. CLARITY-processed retinas were immunostained with antibodies for retinal neurons, glial cells, and synaptic proteins, mounted in a refractive index matching solution, and imaged. Our data demonstrate that CLARITY&#160;can improve the quality of standard immunohistochemical staining and imaging for retinal neurons and glial cells in whole-mount preparation. For instance, 3D resolution of fine axon-like and dendritic structures of dopaminergic amacrine cells were much improved by CLARITY. Compared to non-processed whole-mount retinas, CLARITY can reveal immunostaining for synaptic proteins such as postsynaptic density protein 95. Our results show that CLARITY renders the retina more optically transparent after the removal of lipids and preserves fine structures of retinal neurons and their proteins, which can be routinely used for obtaining high-resolution imaging of retinal neurons and their subcellular structures in whole-mount preparation.

Here we present a protocol to adapt the CLARITY method of the brain tissues for whole-mount retinas to improve the quality of standard immunohistochemical staining and high-resolution imaging of retinal neurons and their subcellular structures.

使用清晰组织清除方法对全山视网膜进行免疫染色

The tissue hydrogel delipidation method (CLARITY), originally developed by the Deisseroth laboratory, has been modified and widely used for immunostaining ...