KIST格兰特（2E21510），教育部国家荣誉的科学家计划，并通过韩国国家研究基金会原有的技术为脑科学的研究计划，由教育部，科学和技术（2010-0018944）资助支持。

1。摘要只C57BL/6J - 129S4/SvJae混合小鼠（12〜15周龄）麻醉与氯胺酮（120毫克/公斤的IP）和甲苯噻嗪（6毫克/公斤，IP）的鸡尾酒。尾巴或脚趾捏之前检查手术麻醉深度。鼠标是固定的立体定位仪（大卫Kopf仪器，型号902，加州，美国）。头皮切开约25毫米利多卡因减轻疼痛的治疗颅骨暴露在中线。任何头骨碎片是被盐水浸泡过的棉花提示，以提高电极坚持头骨。前囟门和lambda点标记，并在同一水平面。聚酰亚胺为基础的微电极阵列（矿务局阵列）的位置，所以，它的垂直中线符合的头骨中线。前囟门点应设在第 5层的前矿务局阵列中间。使用两个或三个microscrews在边缘的头骨，以帮助确保电极用牙科水泥的位置空空间。为支撑矿务局阵列连接器，我们对头部的中间支撑位置，并用牙科水泥覆盖。固化后，切开皮肤缝合，至少5天的恢复。在恢复过程中，老鼠被安置在个别笼免费获得食物和水。恢复后，进行脑电记录。动物护理，外科手术，并记录程序，按照机构的动物护理和使用委员会的指引，继1992年的韩国实验室动物保护条例"和相关准则的法。在这个手稿矿务局阵列制备蔡描述。等。人请联系作者在视频使用的相同的PBM阵列。 2。程序<ol><li>准备消毒的手术器械和立体定位仪。 </li><li>麻醉与麻醉（例如，氯胺酮/甲苯​​噻嗪，剂量为120和60毫克/公斤的IP，repectively）鼠标。尾巴或脚趾捏之前检查手术麻醉深度。 </li><li>立体定位仪安装到鼠标放入鼠标的耳道，耳酒吧，并拧紧到位。眼睛可覆盖生物相容性果冻（如凡士林），以防止眼睛干燥，手术灯。保湿鼠标的头部，用70％乙醇后，剃光头皮毛。 </li><li>切开头皮约25毫米的头部。这是可取的，适用2％的少量利多卡因头皮下的盐酸。捏皮肤微夹钳（18052-03，精细的科学工具（美国），公司​​，加州，美国），以保持切口敞开。 </li><li>擦拭用盐水浸泡棉签提高矿务局阵列坚持颅骨其余的头骨上的任何组织碎片。使用去除膜的残留过氧化氢也是有效的。 </li><li>前囟和lambda点上找到的头骨，并与石油为基础的记号笔标记。矿务局阵列，所以，它的垂直中线符合图1所示的头骨中线放置。矿务局阵列的前5次用一滴自来水层中间位置的前囟门。从内侧向外侧，为更好地遵守仔细擦拭薄膜电极。为了帮助薄膜电极薄膜电极坚持严格的头骨，应完全干燥的头骨。薄膜电极仍然被固定在头骨上的范德华力。重新定位电极，另一滴自来水是必要的，以防止被撕裂的电极。 <img src="/files/ftp_upload/2562/2562fig1.jpg" alt="图1" /> 图1（左）鼠标头骨矿务局阵列。矿务局阵列anterio后轴相匹配的前囟（BP）和lambda点（LP）之间的线。 （右）上公认的鼠脑表面电极位置。鼠标的解剖结构分割的3 - D的磁共振成像数字地图集在布鲁克海文国家实验室和电极位置生产的成年C57BL/6J小鼠脑数据库，根据立体测量标记。地线（G）和参考（REF）电极标记在右边的图。 </li><li>标记至少有三个开放区域的头骨上的位置，然后在这些地点使用支持microscrews定位一个牙钻的孔。这些microscrews的stanchion作用和机械支持的头骨上的牙科水泥。在钻井过程中需要格外小心。由于小鼠颅骨的薄层组成，出血可发生在钻井过程中轻松。出血发生时，停钻成孔止血粉少量申请。 </li><li>应用牙科丙烯酸水泥（牙科顶点，顶点自固化，荷兰），覆盖的PBM的阵列，锚钉，而其余的头骨。准备在胶和粘性彗星的牙科水泥ondition。 </li><li>一个刚性棒作为一个支柱上使用强力胶头（例如，乐泰，汉高乐泰公司，都柏林，爱尔兰）的中间位置。放在后面的连接器平面双面胶带。操纵矿务局阵列连接器连接支柱的一角。头骨和连接器之间的垂直距离应超过5毫米。棒连接器的平面和提示使用强力胶水粘更好的附着力。 </li><li>确保彻底治愈，牙科水泥。然后申请在头骨的牙齿丙烯酸水泥和安全颅骨的连接器连接器平面的背面。 </li><li>删除微夹钳。缝合切开皮肤消毒缝合纤维。从设备中取出鼠标。你可以申请额外的牙科丙烯酸水泥盾暴露颅骨。周围的水泥和缝合皮肤应用抗生素（fucidin软膏，东和制药，韩国）。 </li><li>鼠标放置在消毒笼，直到它恢复其原来的体重（约5至7天）。建议不作任何笼交配。 </li><li>恢复后，由一个轻量级的印刷电路板（PCB）连接器连接到一个多通道脑电放大器系统，在自己的笼子里（Synamps 2），美国北卡罗莱纳州的流浆箱的连接鼠标。连接线应慎重对待，因此它不纠缠不清。 </li><li>经过1小时的习惯，进行脑电记录。扫描4.4。 （美国北卡罗莱纳州）在本实验中使用采集软件。一个典型的时间跟踪鼠标脑电图的设置如图2所示。建议记录的视频与鼠标的行为。 </li><li> MATLAB（MathWorks公司，纳提克，马萨诸塞州，美国）获得的信号可以分析产生的地形脑图。鼠脑的脑电功率图的示例拓扑图如图3所示。 </li></ol> 3。代表性的成果如果上述手术过程中正确地进行，将成功地执行了40个频道的鼠标脑电记录，无不良渠道。下图显示的代表性原始脑电图跟踪和地形图。 <img src="/files/ftp_upload/2562/2562fig2.jpg" alt="图2" /> 图2:一个脑电图经历的情况下通过注射γ-丁内酯（GBL，经口50mg/kg，IP）致痫样本跟踪。红色条表示在失神发作棘波放电（SWDS）。水平和垂直轴表示时间和电压分别。在图中定义的通道蒙太奇数如下。 1（右）。 <img src="/files/ftp_upload/2562/2562fig3.jpg" alt="图3" /> 图3。地形图显示2-6赫兹配电。在彩条的值代表的势力地图的颜色规模。在棘波和γ-丁内酯（GBL，经口50mg/kg，IP），大额顶皮层区域的力量诱导显着的排放（SWDS）相比增加GBL注射前（基线）的权力。

<ol>
	<li>Choi, J. H., Koch, K. P., Poppendieck, W., Lee, M., Shin, H. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High+resolution+Electroencephalography+in+Freely+Moving+Mice.">High resolution Electroencephalography in Freely Moving Mice.</a> J Neurophysiol. , .</li><li>Lee, M., Shin, H. S., Choi, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simultaneous+recording+of+brain+activity+and+functional+connectivity+in+the+mouse+brain.">Simultaneous recording of brain activity and functional connectivity in the mouse brain.</a> Conf Proc IEEE Eng Med Biol Soc. , 2934-2936 (2009).</li></ol>

在这里，我们报告，手术及录音的程序，以获得自由活动的小鼠高密度的脑电图。这种方法使我们获得脑功能的地图，从小鼠模型，研究自发的有节奏的1,2或事件相关脑活动的分子机制。 矿务局阵列中的所有渠道收购是一个必要条件，以获得整个大脑映射，但是为每个通道的安全接触是非常具有挑战性的。为了保持一致的联系渠道颅骨，下面的程序是重要的。首先，任何组织头骨碎片可能会设置除了从头骨的电极。因此，它是非常重要的，以消除所有的组织碎片，并用消毒棉签干颅骨表面。其次，矿务局阵列的能力，能坚持到头骨表面的范德华力。建议申请前申请矿务局阵列头骨，头骨建立水层下降的自来水。第三，正确的牙科水泥粘度是非常重要的。如果是过流鼻涕，有可能建立一个绝缘层，下矿务局阵列。如果过于密集，牙科水泥将很容易从头骨，由于其自​​身的重量土崩瓦解。最后，至少有三个配套microscrews应植入的头骨，使牙科水泥紧密结合的头骨。支持microscrews托起牙科水泥和安全的探头，这是必不可少的长期研究的。 由于其尺寸和重量成年小鼠的慢性研究矿务局阵列的应用将受到限制。根据我们的经验，超过25克重的老鼠已经足够使用。作为进一步的应用，通过我们矿务局阵列中的分支之间的空空间，可能引入二次电极记录或刺激的目的。

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high-density eeg recordings of the freely moving mice using polyimide-based microelectrode

脑电图（EEG）显示的一个大型神经元群体的平均的电活动。它被广泛用来作为一种无创性脑认知神经科学的监测工具以及神经内科癫痫和睡眠障碍的诊断工具。然而，脑电图节律产生的基本机制仍然是下的面纱。最近推出的聚酰亚胺为基础的微电极脑电图 1高分辨率鼠标（矿务局阵列）的试验之一，回答基于丰富的遗传资源，在小鼠模型包含复杂的脑电图生成过程分析脑电信号的神经生理问题。这nanofabricated矿务局阵列对小鼠的头骨中的应用是一个有效的工具，收集大型的转基因小鼠的大脑活动和容纳结合与行为，以确定一定的脑电节律的神经关联。但它的超薄厚度和分叉结构导致矿务局阵列处理和植入的麻烦。在视频中，矿务局阵列鼠标头骨植入的准备和手术步骤描述一步一步来。还提供了处理和手术技巧，以帮助研究人员成功植入。

Watch this Scientific Journal Video about High-density EEG Recordings of the Freely Moving Mice using Polyimide-based Microelectrode at JoVE.com

High-density EEG Recordings of the Freely Moving Mice using Polyimide-based Microelectrode

在这篇文章中，我们介绍了手术过程和处理超薄的聚酰亚胺为基础的微电极阵列（PBM阵列）收购的高密度脑电图（EEG）在小鼠模​​型脑照相鼠标颅骨植入的提示。

高密度的自由移动，使用基于聚酰亚胺的微电极的小鼠的脑电图记录

Electroencephalogram (EEG) indicates the averaged electrical activity of the neuronal populations on a large-scale level. It is widely utilized as a ...

high-density-eeg-recordings-freely-moving-mice-using-polyimide-based

Nanyang Technological University

Research

JoVE Journal

Neuroscience

36.9K 次观看.  Korea Institute of Science and Technology (KIST). 在这篇文章中，我们介绍了手术过程和处理超薄的聚酰亚胺为基础的微电极阵列（PBM阵列）收购的高密度脑电图（EEG）在小鼠模​​型脑照相鼠标颅骨植入的提示。

视频: High-density EEG Recordings of the Freely Moving Mice using Polyimide-based Microelectrode

Combining Computer Game-Based Behavioural Experiments With High-Density EEG and Infrared Gaze Tracking

Experimental paradigms are valuable insofar as the timing and other parameters of their stimuli are well specified and controlled, and insofar as they yield data relevant to the cognitive processing that occurs under ecologically valid conditions. These two goals often are at odds, since well controlled stimuli often are too repetitive to sustain subjects' motivation. Studies employing electroencephalography (EEG) are often especially sensitive to this dilemma between ecological validity and experimental control: attaining sufficient signal-to-noise in physiological averages demands large numbers of repeated trials within lengthy recording sessions, limiting the subject pool to individuals with the ability and patience to perform a set task over and over again. This constraint severely limits researchers' ability to investigate younger populations as well as clinical populations associated with heightened anxiety or attentional abnormalities. Even adult, non-clinical subjects may not be able to achieve their typical levels of performance or cognitive engagement: an unmotivated subject for whom an experimental task is little more than a chore is not the same, behaviourally, cognitively, or neurally, as a subject who is intrinsically motivated and engaged with the task. A growing body of literature demonstrates that embedding experiments within video games may provide a way between the horns of this dilemma between experimental control and ecological validity. The narrative of a game provides a more realistic context in which tasks occur, enhancing their ecological validity (Chaytor &amp; Schmitter-Edgecombe, 2003). Moreover, this context provides motivation to complete tasks. In our game, subjects perform various missions to collect resources, fend off pirates, intercept communications or facilitate diplomatic relations. In so doing, they also perform an array of cognitive tasks, including a Posner attention-shifting paradigm (Posner, 1980), a go/no-go test of motor inhibition, a psychophysical motion coherence threshold task, the Embedded Figures Test (Witkin, 1950, 1954) and a theory-of-mind (Wimmer &amp; Perner, 1983) task. The game software automatically registers game stimuli and subjects' actions and responses in a log file, and sends event codes to synchronise with physiological data recorders. Thus the game can be combined with physiological measures such as EEG or fMRI, and with moment-to-moment tracking of gaze. Gaze tracking can verify subjects' compliance with behavioural tasks (e.g. fixation) and overt attention to experimental stimuli, and also physiological arousal as reflected in pupil dilation (Bradley et al., 2008). At great enough sampling frequencies, gaze tracking may also help assess covert attention as reflected in microsaccades - eye movements that are too small to foveate a new object, but are as rapid in onset and have the same relationship between angular distance and peak velocity as do saccades that traverse greater distances. The distribution of directions of microsaccades correlates with the (otherwise) covert direction of attention (Hafed &amp; Clark, 2002).

Procedures for recording high-density EEG and gaze data during computer game-based cognitive tasks are described. Using a video game to present cognitive tasks enhances ecological validity without sacrificing experimental control.

结合电玩与高密度的脑电图和红外视线跟踪基于行为的实验

Experimental paradigms are valuable insofar as the timing and other parameters of their stimuli are well specified and controlled, and insofar as they ...

科研

神经科学

Recording Single Neurons' Action Potentials from Freely Moving Pigeons Across Three Stages of Learning

While the subject of learning has attracted immense interest from both behavioral and neural scientists, only relatively few investigators have observed single-neuron activity while animals are acquiring an operantly conditioned response, or when that response is extinguished. But even in these cases, observation periods usually encompass only a single stage of learning, i.e. acquisition or extinction, but not both (exceptions include protocols employing reversal learning; see Bingman&#160;et al.1 for an example). However, acquisition and extinction entail different learning mechanisms and are therefore expected to be accompanied by different types and/or loci of neural plasticity.
Accordingly, we developed a behavioral paradigm which institutes three stages of learning in a single behavioral session and which is well suited for the simultaneous recording of single neurons&#39; action potentials. Animals are trained on a single-interval forced choice task which requires mapping each of two possible choice responses to the presentation of different novel visual stimuli (acquisition). After having reached a predefined performance criterion, one of the two choice responses is no longer reinforced (extinction). Following a certain decrement in performance level, correct responses are reinforced again (reacquisition). By using a new set of stimuli in every session, animals can undergo the acquisition-extinction-reacquisition process repeatedly. Because all three stages of learning occur in a single behavioral session, the paradigm is ideal for the simultaneous observation of the spiking output of multiple single neurons. We use pigeons as model systems, but the task can easily be adapted to any other species capable of conditioned discrimination learning.

Learning new stimulus-response associations engages a wide range of neural processes which are ultimately reflected in changing spike output of individual neurons. Here we describe a behavioral protocol allowing for the continuous registration of single-neuron activity while animals acquire, extinguish, and reacquire a conditioned response within a single experimental session.

自由移动鸽子穿越三个阶段学习记录单个神经元&#39;动作电位

While the subject of learning has attracted immense interest from both behavioral and neural scientists, only relatively few investigators have observed ...

Recording EEG in Freely Moving Neonatal Rats Using a Novel Method

EEG is a useful method to detect electrical activity in the brain. Moreover, it is a widely used diagnostic tool for various neurological conditions, such as epilepsy and neurodegenerative disorders. However, it is technically difficult to obtain EEG recordings in neonates as it requires specialized handling and great care. Here, we present a novel method to record EEG in neonatal rat pups (P8-P15). We designed a simple and reliable electrode using computer pin loci; it can be easily implanted into the skull of a rat pup to record high-quality EEG signals in the normal and epileptic brain. Pups were given an intraperitoneal (i.p.) injection of the neurotoxin kainic acid (KA) to induce epileptic seizures. The surgical implantation performed in this procedure is less expensive than other EEG procedures for neonates. This method allows one to record high-quality and stable EEG signals for more than 1 week. Furthermore, this procedure can also be applied to adult rats and mice to study epilepsy or other neurological disorders.

Here, we introduce a novel technique designed to record electroencephalography (EEG) in freely moving neonatal epileptic pups and describe its procedures, features, and applications. This method allows one to record EEG for more than 1 week.

使用新颖的方法在自由移动的新生大鼠中记录脑电图

EEG is a useful method to detect electrical activity in the brain. Moreover, it is a widely used diagnostic tool for various neurological conditions, such ...

Visual Evoked Potential Recordings in Mice Using a Dry Non-invasive Multi-channel Scalp EEG Sensor

For scalp EEG research environments with laboratory mice, we designed a dry-type 16 channel EEG sensor which is non-invasive, deformable, and re-usable because of the plunger-spring-barrel structural facet and mechanical strengths resulting from metal materials. The whole process for acquiring the VEP responses in vivo from a mouse consists of four steps: (1) sensor assembly, (2) animal preparation, (3) VEP measurement, and (4) signal processing. This paper presents representative measurements of VEP responses from multiple mice with a submicro-voltage signal resolution and sub-hundred millisecond temporal resolution. Although the proposed method is safer and more convenient compared to other previously reported animal EEG acquiring methods, there are remaining issues including how to enhance the signal-to-noise ratio and how to apply this technique with freely moving animals. The proposed method utilizes easily available resources and shows a repetitive VEP response with a satisfactory signal quality. Therefore, this method could be utilized for longitudinal experimental studies and reliable translational research exploiting non-invasive paradigms.

We designed a dry-type 16 channel EEG sensor which is non-invasive, deformable, and re-usable. This paper describes the whole process from manufacturing the proposed EEG electrode to signal processing of visual evoked potential (VEP) signals measured on a mouse scalp using a dry non-invasive multi-channel EEG sensor.

干无创多通道头皮脑电传感器在小鼠视觉诱发电位记录中的应用

For scalp EEG research environments with laboratory mice, we designed a dry-type 16 channel EEG sensor which is non-invasive, deformable, and re-usable ...

Simultaneous Recordings of Cortical Local Field Potentials, Electrocardiogram, Electromyogram, and Breathing Rhythm from a Freely Moving Rat

Monitoring the physiological dynamics of the brain and peripheral tissues is necessary for addressing a number of questions about how the brain controls body functions and internal organ rhythms when animals are exposed to emotional challenges and changes in their living environments. In general experiments, signals from different organs, such as the brain and the heart, are recorded by independent recording systems that require multiple recording devices and different procedures for processing the data files. This study describes a new method that can simultaneously monitor electrical biosignals, including tens of local field potentials in multiple brain regions, electrocardiograms that represent the cardiac rhythm, electromyograms that represent awake/sleep-related muscle contraction, and breathing signals, in a freely moving rat. The recording configuration of this method is based on a conventional micro-drive array for cortical local field potential recordings in which tens of electrodes are accommodated, and the signals obtained from these electrodes are integrated into a single electrical board mounted on the animal's head. Here, this recording system was improved so that signals from the peripheral organs are also transferred to an electrical interface board. In a single surgery, electrodes are first separately implanted into the appropriate body parts and the target brain areas. The open ends of all of these electrodes are then soldered to individual channels of the electrical board above the animal's head so that all of the signals can be integrated into the single electrical board. Connecting this board to a recording device allows for the collection of all of the signals into a single device, which reduces experimental costs and simplifies data processing, because all data can be handled in the same data file. This technique will aid the understanding of the neurophysiological correlates of the associations between central and peripheral organs.

This study introduces a method for the simultaneous recording of local field potentials in the brain, electrocardiograms, electromyograms, and breathing signals of a freely moving rat. This technique, which reduces experimental costs and simplifies data analysis, will contribute to the understanding of the interactions between the brain and peripheral organs.

自由运动大鼠皮质局部电位、心电图、肌电图和呼吸节律的同步记录

Monitoring the physiological dynamics of the brain and peripheral tissues is necessary for addressing a number of questions about how the brain controls ...

Noninvasive EEG Recordings from Freely Moving Piglets

The method allows the recording of high-quality electroencephalograms (EEGs) from freely moving piglets directly in the pigpen. We use a one-channel telemetric electroencephalography system in combination with standard self-adhesive hydrogel electrodes. The piglets are calmed down without the use of sedatives. After their release into the pigpen, the piglets behave normally&#8212;they drink and sleep in the same cycle as their siblings. Their sleep phases are used for the EEG recordings.

Here, we present a protocol to record telemetric electroencephalograms (EEGs) from freely moving piglets directly in the pigpen without the use of a sedative, making it possible to record typical EEG patterns during non-REM sleep, like spindle bursts.

自由移动仔猪的无创脑电图记录

The method allows the recording of high-quality electroencephalograms (EEGs) from freely moving piglets directly in the pigpen. We use a one-channel ...

Microdialysis of Excitatory Amino Acids During EEG Recordings in Freely Moving Rats

Microdialysis is a well-established neuroscience technique that correlates the changes of neurologically active substances diffusing into the brain interstitial space with the behavior and/or with the specific outcome of a pathology (e.g., seizures for epilepsy). When studying epilepsy, the microdialysis technique is often combined with short-term or even long-term video-electroencephalography (EEG) monitoring to assess spontaneous seizure frequency, severity, progression and clustering. The combined microdialysis-EEG is based on the use of several methods and instruments. Here, we performed in vivo microdialysis and continuous video-EEG recording to monitor glutamate and aspartate outflow over time, in different phases of the natural history of epilepsy in a rat model. This combined approach allows the pairing of changes in the neurotransmitter release with specific stages of the disease development and progression. The amino acid concentration in the dialysate was determined by liquid chromatography. Here, we describe the methods and outline the principal precautionary measures one should take during in vivo microdialysis-EEG, with particular attention to the stereotaxic surgery, basal and high potassium stimulation during microdialysis, depth electrode EEG recording and high-performance liquid chromatography analysis of aspartate and glutamate in the dialysate. This approach may be adapted to test a variety of drug or disease induced changes of the physiological concentrations of aspartate and glutamate in the brain. Depending on the availability of an appropriate analytical assay, it may be further used to test different soluble molecules when employing EEG recording at the same time.

Here, we describe a method for in vivo microdialysis to analyze aspartate and glutamate release in the ventral hippocampus of epileptic and non-epileptic rats, in combination with EEG recordings. Extracellular concentrations of aspartate and glutamate may be correlated with the different phases of the disease.

自由运动大鼠脑电图记录过程中兴奋氨基酸的微透析

Microdialysis is a well-established neuroscience technique that correlates the changes of neurologically active substances diffusing into the brain ...

Long-term Sensory Conflict in Freely Behaving Mice

Long-term sensory conflict protocols are a valuable means of studying motor learning. The presented protocol produces a persistent sensory conflict for experiments aimed at studying long-term learning in mice. By permanently wearing a device fixed on their heads, mice are continuously exposed to a sensory mismatch between visual and vestibular inputs while freely moving in home cages. Therefore, this protocol readily enables the study of the visual system and multisensory interactions over an extended timeframe that would not be accessible otherwise. In addition to lowering the experimental costs of long-term sensory learning in naturally behaving mice, this approach accommodates the combination of in vivo and in vitro experiments. In the reported example, video-oculography is performed to quantify the vestibulo-ocular reflex (VOR) and optokinetic reflex (OKR) before and after learning. Mice exposed to this long-term sensory conflict between visual and vestibular inputs presented a strong VOR gain decrease but exhibited few OKR changes. Detailed steps of device assembly, animal care, and reflex measurements are hereby reported.

The presented protocol produces a persistent sensory conflict for experiments aimed at studying long-term learning. By permanently wearing a fixed device on their heads, mice are continuously exposed to a sensory mismatch between visual and vestibular inputs while freely moving in home cages.

自由生育小鼠的长期感觉冲突

Long-term sensory conflict protocols are a valuable means of studying motor learning. The presented protocol produces a persistent sensory conflict for ...

Hybrid Microdrive System with Recoverable Opto-Silicon Probe and Tetrode for Dual-Site High Density Recording in Freely Moving Mice

Multi-regional neural recordings can provide crucial information to understanding fine-timescale interactions between multiple brain regions. However, conventional microdrive designs often only allow use of one type of electrode to record from single or multiple regions, limiting the yield of single-unit or depth profile recordings. It also often limits the ability to combine electrode recordings with optogenetic tools to target pathway and/or cell type specific activity. Presented here is a hybrid microdrive array for freely moving mice to optimize yield and a description of its fabrication and reuse of the microdrive array. The current design employs nine tetrodes and one opto-silicon probe implanted in two different brain areas simultaneously in freely moving mice. The tetrodes and the opto-silicon probe are independently adjustable along the dorsoventral axis in the brain to maximize the yield of unit and oscillatory activities. This microdrive array also incorporates a set-up for light, mediating optogenetic manipulation to investigate the regional- or cell type-specific responses and functions of long-range neural circuits. In addition, the opto-silicon probe can be safely recovered and reused after each experiment. Because the microdrive array consists of 3D-printed parts, the design of microdrives can be easily modified to accommodate various settings. First described is the design of the microdrive array and how to attach the optical fiber to a silicon probe for optogenetics experiments, followed by fabrication of the tetrode bundle and implantation of the array into a mouse brain. The recording of local field potentials and unit spiking combined with optogenetic stimulation also demonstrate feasibility of the microdrive array system in freely moving mice.

This protocol describes the construction of a hybrid microdrive array that allows implantation of nine independently adjustable tetrodes and one adjustable opto-silicon probe in two brain regions in freely moving mice. Also demonstrated is a method for safely recovering and reusing the opto-silicon probe for multiple purposes.

混合微驱系统,带可回收的光电探头和Tetrode,用于自由移动小鼠的双位点高密度记录

Multi-regional neural recordings can provide crucial information to understanding fine-timescale interactions between multiple brain regions. However, ...

Multi-Fiber Photometry to Record Neural Activity in Freely-Moving Animals

Recording the activity of a group of neurons in a freely-moving animal is a challenging undertaking. Moreover, as the brain is dissected into smaller and smaller functional subgroups, it becomes paramount to record from projections and/or genetically-defined subpopulations of neurons. Fiber photometry is an accessible and powerful approach that can overcome these challenges. By combining optical and genetic methodologies, neural activity can be measured in deep brain structures by expressing genetically-encoded calcium indicators, which translate neural activity into an optical signal that can be easily measured. The current protocol details the components of a multi-fiber photometry system, how to access deep brain structures to deliver and collect light, a method to account for motion artifacts, and how to process and analyze fluorescent signals. The protocol details experimental considerations when performing single and dual color imaging, from either single or multiple implanted optic fibers.

This protocol details how to implement and perform multi-fiber photometry recordings, how to correct for calcium-independent artifacts, and important considerations for dual-color photometry imaging.

多纤维光测量记录自由移动动物的神经活动

Recording the activity of a group of neurons in a freely-moving animal is a challenging undertaking. Moreover, as the brain is dissected into smaller and ...