Name: in vivo microdialysis method to collect large extracellular proteins from brain interstitial fluid with high-molecular weight cut-off probes
Uploaded: 2018-09-26T12:30:00
Description: Watch this Scientific Journal Video about In Vivo Microdialysis Method to Collect Large Extracellular Proteins from Brain Interstitial Fluid with High-molecular Weight Cut-off Probes at JoVE.com

这项工作得到了来自下个和年轻科学家 (B) (16K20969) 资助的 "对创新领域 (脑蛋白质老化和痴呆控制) (15H01552) 进行科学研究的资助" 的支持。作者感谢 David m. Holtzman 博士和 Cirrito 博士在这一方法的发展过程中提出的技术建议。

所有动物研究都由东京大学医学研究生院机构动物护理和使用委员会审查和批准。
1. 手术前的程序
<ol><li>在开始手术前, 用70% 乙醇擦拭所有的东西以保持不育的条件。推荐使用加热垫的热支持。</li>
	<li>麻醉对小鼠进行腹腔注射水合氯醛 (400 毫克/千克)。通过执行脚趾夹来确认麻醉。推荐使用 meloxicam SR 在感应和丁丙诺啡恢复出价至少24小时。</li>
	<li>用手术剪刀刮头发。用?...

<ol>
	<li>Lei, Y., Han, H., Yuan, F., Javeed, A., Zhao, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+brain+interstitial+system:+Anatomy,+modeling,+in+vivo+measurement,+and+applications.">The brain interstitial system: Anatomy, modeling, in vivo measurement, and applications.</a> Progress in Neurobiology. 157, 230-246 (2017).</li><li>Kushikata, T., Hirota, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuropeptide+microdialysis+in+free-moving+animals.">Neuropeptide microdialysis in free-moving animals.</a> Methods in Molecular Biology. 789, 261-269 (2011).</li><li>Cirrito, J. 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D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+measurement+of+apolipoprotein+E+from+the+brain+interstitial+fluid+using+microdialysis.">In vivo measurement of apolipoprotein E from the brain interstitial fluid using microdialysis.</a> Molecular Neurodegeneration. 8 (1), 13 (2013).</li><li>Emmanouilidou, E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=GABA+transmission+via+ATP-dependent+K++channels+regulates+&#945;-synuclein+secretion+in+mouse+striatum.">GABA transmission via ATP-dependent K+ channels regulates &#945;-synuclein secretion in mouse striatum.</a> Brain. 139 (3), 871-890 (2016).</li><li>Takeda, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Seed-competent+high-molecular-weight+tau+species+accumulates+in+the+cerebrospinal+fluid+of+Alzheimer's+disease+mouse+model+and+human+patients.">Seed-competent high-molecular-weight tau species accumulates in the cerebrospinal fluid of Alzheimer's disease mouse model and human patients.</a> Annals of Neurology. 80 (3), 355-367 (2016).</li><li>Yamada, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+Microdialysis+of+brain+interstitial+fluid+for+the+determination+of+extracellular+tau+levels.">In vivo Microdialysis of brain interstitial fluid for the determination of extracellular tau levels.</a> Methods in Molecular Biology. 1523, 285-296 (2017).</li><li>Kang, J. -. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Amyloid-&#946;+dynamics+are+regulated+by+orexin+and+the+sleep-wake+cycle.">Amyloid-&#946; dynamics are regulated by orexin and the sleep-wake cycle.</a> Science. 326 (November), 1005-1008 (2009).</li><li>Cirrito, J. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synaptic+activity+regulates+interstitial+fluid+amyloid-beta+levels+in+vivo.">Synaptic activity regulates interstitial fluid amyloid-beta levels in vivo.</a> Neuron. 48 (6), 913-922 (2005).</li><li>Bero, A. W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuronal+activity+regulates+the+regional+vulnerability+to+amyloid-&#946;+deposition.">Neuronal activity regulates the regional vulnerability to amyloid-&#946; deposition.</a> Nature Neuroscience. 14 (6), 750-756 (2011).</li><li>Yamada, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuronal+activity+regulates+extracellular+tau+in+vivo.">Neuronal activity regulates extracellular tau in vivo.</a> Journal of Experimental Medicine. 211 (3), 387-393 (2014).</li><li>Pooler, A. M., Phillips, E. C., Lau, D. H. W., Noble, W., Hanger, D. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Physiological+release+of+endogenous+tau+is+stimulated+by+neuronal+activity.">Physiological release of endogenous tau is stimulated by neuronal activity.</a> EMBO Reports. 14 (4), 389-394 (2013).</li><li>Castellano, J. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+apoE+isoforms+differentially+regulate+brain+amyloid-&#946;+peptide+clearance.">Human apoE isoforms differentially regulate brain amyloid-&#946; peptide clearance.</a> Science Translational Medicine. 3 (89), 89ra57 (2011).</li><li>Yamada, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+of+in+vivo+turnover+of+tau+in+a+mouse+model+of+tauopathy.">Analysis of in vivo turnover of tau in a mouse model of tauopathy.</a> Molecular Neurodegeneration. 10 (1), 55 (2015).</li><li>Taylor, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Investigating+local+and+long-range+neuronal+network+dynamics+by+simultaneous+optogenetics,+reverse+microdialysis+and+silicon+probe+recordings+in+vivo.">Investigating local and long-range neuronal network dynamics by simultaneous optogenetics, reverse microdialysis and silicon probe recordings in vivo.</a> Journal of Neuroscience Methods. 235, 83-91 (2014).</li></ol>

高分子量切断膜的微透析必须采用推拉方式操作, 因此流速准确、恒定是至关重要的。流动速率的不准确性可能是气泡产生的原因和样品浓度不一致。如果流程不一致, 请检查所有连接是否有泄漏。如果问题仍然存在, 可能需要重新启动新的探头和油管。
Microdilaysis 探头通过灌注缓冲液不断灌注。因此, 细胞外分子没有足够的时间在灌注缓冲液中达到完全平衡。因此, 透析液中?...

该方法包括总脑容积的 15-20%, 并提供了一个关键的微环境对信号传导, 基质运输和废物清除1。因此, 收集活体动物的能力将对各种生物过程和疾病机制带来更大的影响。体内微透析是从清醒的、自由移动的动物中取样和量化细胞外分子的为数不多的方法之一, 从而在神经科学研究领域2、3中成为有用的工具。在该方法中, 透膜的微透析探针插入脑中, 以相对缓慢的流速 (0.1-5 µL/分钟) 灌流灌注缓冲液。在此灌注过程中, 细胞外分子根据浓度梯度被动扩散到探针中, 并作为透析液收集。虽然本文的重点是对脑部标本的方法, 但无论是原则还是方法, 都可以通过适当的修改在必要的情况下应用于其他器官。
微透析在二十世纪六十年代代初首次被使用, 从那时起, 它被广泛用于收集小分子, 包括氨基酸或脑内神经递质。然而, 最近的商业可用性微透析探针与高分子量的切割膜 (100 kDa-3 MDa) 已扩大其应用到相对较大的蛋白质, 以及4,5,6 ,7。使用这些探针的研究结果发现, 长时间被认为是专属细胞质的头或α synuclein 的蛋白质也在生理上存在于4,5,8。
使用微透析探针与大型切断膜 (通常超过 1000 kDa) 的难点之一是, 由于探针中积聚的内压, 它们更容易受到超滤液的损耗。这里使用的微透析探针有一个独特的结构来避免这个问题。压力不会被建立由于这个结构, 因此微透析与这些探针应该操作在 "推挤-拉扯" 方式使用注射器泵灌注探针 (= 推挤) 和滚筒/蠕动泵收集从探针出口的透析器(= 拉)9 (虽然它需要推挤和拉扯泵, 由于压力取消透气孔存在于探针, 系统在技术上仅由拉扯泵驱动)。本文从引导套管植入的立体定向手术入手, 介绍了如何建立微透析线, 以便通过微透析探针与 1000 kDa 膜分离来采集。

<table>
	<tbody>
		<tr>
			<td>The Univentor 820 Microsampler</td>
			<td>Univentor</td>
			<td>8303002</td>
			<td>Refrigerated fraction collector</td>
		</tr>
		<tr>
			<td>Syringe pump</td>
			<td>KD scientific</td>
			<td>KDS-101</td>
			<td></td>
		</tr>
		<tr>
			<td>Roller pump</td>
			<td>Eicom microdialysis</td>
			<td>ERP-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Raturn Stand-Alone System</td>
			<td>BASi</td>
			<td>MD-1409</td>
			<td>Free-moving system</td>
		</tr>
		<tr>
			<td>Dual species cage kit</td>
			<td>BASi</td>
			<td>CX-1600</td>
			<td></td>
		</tr>
		<tr>
			<td>AtmosLM Microdialysis probe (shaft length 8 mm, membrane length 2 mm)</td>
			<td>Eicom microdialysis</td>
			<td>PEP-8-02</td>
			<td>Shaft length for a probe, a guide, a dumy probe and a stereotaxic adaptor should be identical.</td>
		</tr>
		<tr>
			<td>Microdialysis guide (shaft length 8 mm)</td>
			<td>Eicom microdialysis</td>
			<td>PEG-8</td>
			<td></td>
		</tr>
		<tr>
			<td>Microdialysis dummy probe (shaft length 8 mm)</td>
			<td>Eicom microdialysis</td>
			<td>PED-8</td>
			<td></td>
		</tr>
		<tr>
			<td>Bone screw</td>
			<td>BASi</td>
			<td>MD-1310</td>
			<td></td>
		</tr>
		<tr>
			<td>Super bond C&#38;B set</td>
			<td>Sunmedical</td>
			<td></td>
			<td>Dental cement</td>
		</tr>
		<tr>
			<td>Small animal Stereotaxic Instrument with digital display console</td>
			<td>Kopf</td>
			<td>Model 940</td>
			<td>Stereotaxic apparatus</td>
		</tr>
		<tr>
			<td>Mouse and neonatal rat adaptor</td>
			<td>Stoelting</td>
			<td>51625</td>
			<td></td>
		</tr>
		<tr>
			<td>Standard Ear Bars and Rubber Tips for Mouse Stereotaxic</td>
			<td>Stoelting</td>
			<td>51648</td>
			<td></td>
		</tr>
		<tr>
			<td>Albumin solution from bovine serum</td>
			<td>Sigma</td>
			<td>A7284-50ML</td>
			<td>30% BSA solution</td>
		</tr>
		<tr>
			<td>FEP tubing (70 cm)</td>
			<td>Eicom microdialysis</td>
			<td>JF-10-70</td>
			<td>Internal volume = 0.5 &#181;L/cm</td>
		</tr>
		<tr>
			<td>Teflon tubing (50 cm)</td>
			<td>Eicom microdialysis</td>
			<td>JT-10-50</td>
			<td>Internal volume = 0.08 &#181;L/cm</td>
		</tr>
		<tr>
			<td>Byton tube</td>
			<td>Eicom microdialysis</td>
			<td>JB-30</td>
			<td></td>
		</tr>
		<tr>
			<td>Intramedic luer stab adaptor 23G</td>
			<td>BD</td>
			<td>427565</td>
			<td>Blunt end needle</td>
		</tr>
		<tr>
			<td>Roller tube</td>
			<td>Eicom microdialysis</td>
			<td>RT-5S</td>
			<td>Internal volume = 4 &#181;L</td>
		</tr>
		<tr>
			<td>Cap nut</td>
			<td>Eicom microdialysis</td>
			<td>AC-5</td>
			<td></td>
		</tr>
		<tr>
			<td>0.25 mL microcentrifuge tube with cap</td>
			<td>QSP</td>
			<td>503-Q</td>
			<td>Tubes for fraction collector</td>
		</tr>
		<tr>
			<td>Sterotaxic adaptor (shaft length 8 mm)</td>
			<td>Eicom microdialysis</td>
			<td>PESG-8</td>
			<td></td>
		</tr>
		<tr>
			<td>Connection needle</td>
			<td>Eicom microdialysis</td>
			<td>RTJ</td>
			<td></td>
		</tr>
		<tr>
			<td>Mouse animal collar</td>
			<td>BASi</td>
			<td>MD-1365</td>
			<td></td>
		</tr>
		<tr>
			<td>High Speed Rotary Micromotor kit</td>
			<td>FOREDOM</td>
			<td>K.1070</td>
			<td>Drill</td>
		</tr>
		<tr>
			<td>Picrotoxin</td>
			<td>Sigma</td>
			<td>P1675</td>
			<td></td>
		</tr>
		<tr>
			<td>Screw driver for bone screws</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Scalpel</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Cotton swab</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical clipper</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

in vivo microdialysis method to collect large extracellular proteins from brain interstitial fluid with high-molecular weight cut-off probes

在体内微透析是一种强大的技术, 收集从清醒, 自由行为的动物基于透析原理。虽然微透析是一种测量相对较小的分子, 包括氨基酸或神经递质的已建立的方法, 它最近也被用来评估大分子的动力学, 利用探针与高分子量切断膜。使用这种探针时, 微透析必须以推拉模式运行, 以避免探头内部积聚的压力。本文提供了分步协议, 包括立体定向手术, 以及如何建立微透析线收集蛋白质从。在微透析中, 药物可以系统地或直接输注到疫苗中。反向微透析技术是直接将化合物注入到其上的一种方法。在微透析灌注缓冲器中加入药物, 可使它们通过探针扩散到。以头蛋白为例, 通过对化学点燃癫痫的反向微透析, 揭示了其水平在刺激神经元活性方面的变化。本文结合其他体内方法, 介绍了微透析技术的优点和局限性。

为了刺激或抑制反向微透析11,12,13, 化学点燃癫痫, GABAA受体拮抗剂或河豚毒素的神经活动, Na+通道阻滞药已被使用。结果表明, 通过神经元活动的增加13,14刺激了头的释放。与以前的观察一致, 当50µM 化学点燃癫痫 (PTX) 通过反向微透析 (参见讨?...

Watch this Scientific Journal Video about In Vivo Microdialysis Method to Collect Large Extracellular Proteins from Brain Interstitial Fluid with High-molecular Weight Cut-off Probes at JoVE.com

In Vivo Microdialysis Method to Collect Large Extracellular Proteins from Brain Interstitial Fluid with High-molecular Weight Cut-off Probes

体内微透析已使大脑间质液中存在的分子从清醒的、自由行为的动物中收集。为了分析该化合物中相对较大的分子, 本文重点研究了用高分子量切断膜探针的微透析协议。

体内高分子量截离法从脑间质液中提取大细胞外蛋白的微透析方法

In vivo microdialysis is a powerful technique to collect ISF from awake, freely-behaving animals based on a dialysis principle. While microdialysis is an ...

这种方法可以帮助回答神经科学研究领域关于信号转导基板传输和废物清除如何在脑外空间发生关键问题。该技术的主要优点是，它允许对唤醒自由移动的动物的间质流体或 ISF 中的大细胞外分子进行采样和定量。在确认对手趾捏没有反应后，从麻醉小鼠的头骨上取下头发，并使用耳条和鼻夹将固定动物固定在适配器上。将软膏涂抹在动物的眼睛上，将适配器放在立体税器上。使用手术刀在头骨上下垂地进行皮肤切口，并在立体轴框架的操纵器上安装钻头。降低钻头，直到它轻轻触摸 lambda，并重置钻头的 DV 坐标为零。然后，将钻头移动到布雷格玛，以将 AP 和 ML 坐标重置为零。将钻头从啤酒从啤酒移到垂直坐标。将钻头降至头骨，然后再次将 DV 坐标设置为零。重复第三个坐标的过程后，在目标坐标处小心钻一个毛刺孔，然后在骨骼的相反侧钻第二个孔。将骨螺钉插入第二个孔，并将锁块从 1.5 毫升微离心管盖放在头骨上，使毛刺孔位于圆内。接下来，在立体声适配器的短臂上放置一个导轨，用盖螺母固定管。将立体声轴的较长臂放在电极夹上，然后将臂连接到立体机械装置的操纵器上。将操纵臂上的 DV 立体轴组件旋转 12 度，然后将导轨杆移动到毛刺孔。然后，慢慢地将导管下到大脑中1.2毫米。将牙科水泥添加到牙冠上，直到导管、骨螺钉和任何裸露的头骨的金属部分被覆盖和固定，让水泥变干。12 到 20 分钟后，从电极夹上卸下立体声轴适配器，卸下盖螺母，然后用虚拟探头更换立体声适配器。然后，重新发毛帽螺母，将鼠标从立体技术装置中释放出来，将鼠标单独放在单独的笼子里。在进行微透析之前，用蒸馏水填充一次性一毫升注射器，并使用维顿管将注射器连接到微透析探针的插座。手动盖住通风孔，轻轻按下注射器柱塞，将探针注入水。确认探针入口中出现水，并且微透析膜表面有泄漏。要激活探针，将探针膜淹没在70-100%乙醇中两秒钟，然后进行第二次蒸馏水输液。将连接针连接到一个入口和一条出口管路，然后用新鲜准备的灌注缓冲液装载一次性三毫升注射器。将装有钝端针头的注射器连接到管子的入口端，并使用注射器泵向整个管内加注液缓冲液。当管已满时，用激活的微透析探头和盖螺母更换入口和出口管之间的连接针，然后将滚管安装到滚轮泵上的出口管中。以每分钟 10 微升启动注射器泵，以每分钟 9.5-9.8 微升启动滚筒泵。现在，将领子放在麻醉导菜植入动物的脖子上，并拆下帽螺母和假探针。缓慢地插入微透析探针，通过导管，并固定盖螺母。然后，将鼠标放在连接到自由移动系统的笼子中，然后用衣领将鼠标系在一起。至少一小时后，依次停止滚筒泵和注射器泵，重新启动泵，注射器泵设置为比滚筒泵快 20%，并将出口管的自由端放在冷冻分数收集器上，以收集大脑 ISF。获得适当的实验体积后，取出探针并像刚才演示的一样处理鼠标恢复。与先前的观察一致，当50微摩尔皮克罗毒素通过反向微透析施用时，如清醒的C57B6J小鼠所证明的，与在车辆对照治疗的动物中测量的水平相比，观察到间质内源性tau水平增加。除了药物给药，微透析还可以与其他体内方法（如脑电图记录或光遗传学）相结合，以回答有关神经元活动如何影响 ISF 中基质浓度的其他问题。

Research

JoVE Journal

Neuroscience

In Vivo Microdialysis Method to Collect Large Extracellular Proteins from Brain Interstitial Fluid with High-molecular Weight Cut-off Probes

Lateral Fluid Percussion: Model of Traumatic Brain Injury in Mice

侧向液压冲击：创伤性脑损伤小鼠模型

Traumatic brain injury (TBI) research has attained renewed momentum due to the increasing awareness of head injuries, which result in morbidity and ...

科研

神经科学

Recording Large-scale Neuronal Ensembles with Silicon Probes in the Anesthetized Rat

在麻醉大鼠的硅探头录制的大型神经元合唱

Large scale electrophysiological recordings from neuronal ensembles offer the opportunity to investigate how the brain orchestrates the wide variety of ...

Large-scale Recording of Neurons by Movable Silicon Probes in Behaving Rodents

移动硅探针的神经行为鼠害的大型记录

A major challenge in neuroscience is linking behavior to the collective activity of neural assemblies. Understanding of input-output relationships of ...

Retrograde Fluorescent Labeling Allows for Targeted Extracellular Single-unit Recording from Identified Neurons In vivo

Retrograde Fluorescent Labeling Allows for Targeted Extracellular Single-unit Recording from Identified Neurons In vivo

逆行荧光染料标记允许有针对性的外单细胞记录技术鉴定神经元<em&gt;在体内</em

The overall goal of this method is to record single-unit responses from an identified population of neurons. In vivo electrophysiological recordings from ...

Functional Neuroimaging Using Ultrasonic Blood-brain Barrier Disruption and Manganese-enhanced MRI

使用超声波血脑屏障破坏和锰增强MRI的脑功能成像

Although mice are the dominant model system for studying the genetic and molecular underpinnings of neuroscience, functional neuroimaging in mice remains ...

Hippocampal Insulin Microinjection and In vivo Microdialysis During Spatial Memory Testing

Hippocampal Insulin Microinjection and In vivo Microdialysis During Spatial Memory Testing

海马胰岛素显微注射和<em在体内</em&gt;微透析过程中空间记忆测试

Glucose metabolism is a useful marker for local neural activity, forming the basis of methods such as 2-deoxyglucose and functional magnetic resonance ...

Design and Fabrication of Ultralight Weight, Adjustable Multi-electrode Probes for Electrophysiological Recordings in Mice

设计和超轻重量的制作，可调节的多电极电生理记录小鼠探头

The number of physiological investigations in the mouse, mus musculus, has experienced a recent surge, paralleling the growth in methods of genetic ...

Rapid In Situ Hybridization using Oligonucleotide Probes on Paraformaldehyde-prefixed Brain of Rats with Serotonin Syndrome

Rapid In Situ Hybridization using Oligonucleotide Probes on Paraformaldehyde-prefixed Brain of Rats with Serotonin Syndrome

快速<em&gt;原位</em使用寡核苷酸探针对大鼠多聚甲醛为前缀的大脑5-羟色胺综合征&gt;杂交

3,4-Methylenedioxymethamphetamine (MDMA; ecstasy) toxicity may cause region-specific changes in serotonergic mRNA expression due to acute serotonin ...

Real-time Iontophoresis with Tetramethylammonium to Quantify Volume Fraction and Tortuosity of Brain Extracellular Space

用四甲基铵进行实时离子电渗疗法，以量化体细胞分数和脑细胞外空间的变性

This review describes the basic concepts and protocol to perform the real-time iontophoresis (RTI) method, the gold-standard to explore and quantify the ...

Improving the Application of High Molecular Weight Biotinylated Dextran Amine for Thalamocortical Projection Tracing in the Rat

高分子量生物素化葡聚糖胺在大鼠丘脑-皮质投影示踪中的应用

High molecular weight biotinylated dextran amine (BDA) has been used as a highly sensitive neuroanatomical tracer for many decades. Since the quality of ...