Name: visualizing and analyzing intracellular transport of organelles and other cargos in astrocytes
Uploaded: 2019-08-28T15:00:00
Description: Watch this Scientific Journal Video about Visualizing and Analyzing Intracellular Transport of Organelles and Other Cargos in Astrocytes at JoVE.com

DNL作为西蒙斯学者得到北卡罗来纳大学教堂山分校（UNC）医学院的支持。TWR得到了UNCPREP赠款R25 GM089569的支持。使用UNC神经科学中心显微镜核心设施的工作部分得到了NIH-NINDS神经科学中心支持赠款P30 NS045892和NIH-NICHD智力和发育障碍研究中心支持赠款U54的支持支持。HD079124。

所有动物程序都是在北卡罗来纳大学教堂山动物护理和使用委员会（IACUC）批准下进行的。
1. 脑解剖和原小鼠星形细胞培养
注：以下协议改编自已发表的方法，遵循麦卡锡和德韦利斯13、14、15开发的原始程序。麦卡锡/德韦利斯（MD）星形细胞的混合细胞培养从产后2⁄4（P2+P4）小鼠皮质?...

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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=Distribution+of+glutamate+transporter+GLAST+in+membranes+of+cultured+astrocytes+in+the+presence+of+glutamate+transport+substrates+and+ATP.">Distribution of glutamate transporter GLAST in membranes of cultured astrocytes in the presence of glutamate transport substrates and ATP.</a> Neurochemistry Research. 34 (10), 1758-1766 (2009).</li><li>Potokar, 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=Cytoskeleton+and+vesicle+mobility+in+astrocytes.">Cytoskeleton and vesicle mobility in astrocytes.</a> Traffic. 8 (1), 12-20 (2007).</li><li>Potokar, 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=Regulation+of+AQP4+surface+expression+via+vesicle+mobility+in+astrocytes.">Regulation of AQP4 surface expression via vesicle mobility in astrocytes.</a> Glia. 61 (6), 917-928 (2013).</li><li>Potokar, M., Lacovich, V., Chowdhury, H. H., Kreft, M., Zorec, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rab4+and+Rab5+GTPase+are+required+for+directional+mobility+of+endocytic+vesicles+in+astrocytes.">Rab4 and Rab5 GTPase are required for directional mobility of endocytic vesicles in astrocytes.</a> Glia. 60 (4), 594-604 (2012).</li><li>Chiu, C. 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(71), e50079 (2013).</li><li>McCarthy, K. D., de Vellis, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preparation+of+separate+astroglial+and+oligodendroglial+cell+cultures+from+rat+cerebral+tissue.">Preparation of separate astroglial and oligodendroglial cell cultures from rat cerebral tissue.</a> The Journal of Cell Biology. 85 (3), 890-902 (1980).</li><li>Tedeschi, B., Barrett, J. N., Keane, R. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Astrocytes+produce+interferon+that+enhances+the+expression+of+H-2+antigens+on+a+subpopulation+of+brain+cells.">Astrocytes produce interferon that enhances the expression of H-2 antigens on a subpopulation of brain cells.</a> The Journal of Cell Biology. 102 (6), 2244-2253 (1986).</li><li>Zala, 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=Vesicular+glycolysis+provides+on-board+energy+for+fast+axonal+transport.">Vesicular glycolysis provides on-board energy for fast axonal transport.</a> Cell. 152 (3), 479-491 (2013).</li><li>. IJ KymoToolBox Available from: <a target="_blank" href="https://github.com/fabricecordelieres/IJ_KymoToolBox">https://github.com/fabricecordelieres/IJ_KymoToolBox</a> (2016)</li></ol>

在这里，我们描述了一种实验性方法，用于在高纯度原生小鼠皮质MD星形细胞中使用延时视频显微镜来表达、可视化和跟踪荧光标记的细胞器和膜蛋白。我们还概述了测量粒子动力学的方法。在原星形细胞中直接显示蛋白质和细胞器动力学，为研究这些细胞体内细胞内迁移的调节提供了一个有力的工具。
上述建立小鼠MD星形细胞培养的方法结合了其他已公布的协议13、14、15的?...

星形细胞是成人中枢神经系统中最丰富的细胞，它们在其中执行独特的发育和静默功能1。星形细胞调节突触发育，通过直接接触前和后发触终端作为三部分突触的一部分，其中包含神经递质受体，传输器和细胞粘附分子，促进突触形成和神经元-天体细胞通信2。此外，星形细胞通过快速去除突触裂口的兴奋神经递质，回收神经递质，并参与突触修剪3，积极控制突触传播，防止神经元兴奋性中毒。,4,5,6.为了启用这些高度交互的功能，星形细胞通过专门的膜蛋白（包括细胞粘附分子、水孢素、离子电池通道）相互之间、与其他胶质细胞和神经元之间通信，神经递质传输器和间隙结分子。星形细胞积极改变这些蛋白质的表面水平，以响应其细胞内和细胞外环境的波动7。此外，线粒体、脂液滴和降解和再循环细胞器的水平和分布的变化调节了能量供应、代谢物可用性和细胞清除过程，这些对星形细胞功能和生存。
膜蛋白和细胞器贩运和星形细胞的定位的动态变化是由运动蛋白和适配器的协同功能促进货物运动8，9。同样，膜蛋白的表面水平是通过内化和回收事件10来调节的。这些货物通过一个复杂的网络，由行为蛋白、微管和可能的中间细丝轨道8进行运输。基于最终结合蛋白1（EB1）的免疫荧光染色的研究表明，在星形细胞束的微管中，微管从围核外辐射，并将其加端向外围11.然而，仍然缺乏对微管和其他细胞骨骼元素使用活细胞成像的组织和极性的全面检查。虽然细胞器和膜蛋白动力学背后的许多机制已经在神经元和其他细胞类型中进行了广泛的研究，但星形细胞中的货运活力却不太了解。我们目前关于星形细胞中蛋白质和细胞器分布变化的大部分知识是基于传统的基于抗体的固定制备标签，这排除了对货物动力学进行精确的空间和时间检查7， 12.
在这里，我们描述了一种在高纯度原鼠星形细胞培养中为活成像标记膜蛋白和细胞器的方法。使用该协议，我们提供了示例，其中我们跟踪绿色荧光蛋白 （GFP） 标记膜蛋白在转染星细胞中的动态定位，包括间隙结蛋白 Connexin 43 （Cx43-GFP） 和兴奋氨基酸运输器 1 （EAAT1-GFP）。我们还描述了使用荧光酸度探针来可视化酸性细胞器，并跟踪其在活星形细胞中的贩运动态。最后，我们演示了如何分析延时数据，以提取和评估单个货物的运输参数。

<table>
	<tbody>
		<tr>
			<td>2.5% Trypsin (10x)</td>
			<td>Gibco</td>
			<td>15090-046</td>
			<td></td>
		</tr>
		<tr>
			<td>Benchtop Centrifuge</td>
			<td>Thermo Scientific</td>
			<td>75-203-637</td>
			<td>Sorvall ST8 Centrifuge</td>
		</tr>
		<tr>
			<td>Cell Culture Grade Water</td>
			<td>Gen Clone</td>
			<td>25-511</td>
			<td></td>
		</tr>
		<tr>
			<td>Cell Culture Microscope</td>
			<td>Zeiss</td>
			<td>WSN-AXIOVERT A1</td>
			<td>Vert.A1 inverted scope, Inverted tissue culture microscope with fluorescent capabilities</td>
		</tr>
		<tr>
			<td>Cytosine Arabinoside</td>
			<td>Sigma</td>
			<td>C1768-100MG</td>
			<td>(AraC) Dissolve lyophilized powder in sterile cell culture grade water to make a 10mM stock, aliquot and freeze for long term storage</td>
		</tr>
		<tr>
			<td>DAPI</td>
			<td>Sigma</td>
			<td>D9542-5MG</td>
			<td>Dissolve lyophilized powder in deionized water to a maximum concentration of 20 mg/ml, heat or sonicate as necessary. Use at 300 nM for counterstaining.</td>
		</tr>
		<tr>
			<td>Dissecting Microscope</td>
			<td>Zeiss</td>
			<td></td>
			<td>Stemi 305</td>
		</tr>
		<tr>
			<td>Dissecting Scissors</td>
			<td>F.S.T</td>
			<td>14558-09</td>
			<td></td>
		</tr>
		<tr>
			<td>Dulbecco&#39;s Modified Eagle Medium</td>
			<td>Gen Clone</td>
			<td>25-500</td>
			<td></td>
		</tr>
		<tr>
			<td>Fetal Bovine Serum</td>
			<td>Gemini</td>
			<td>100-106</td>
			<td>Heat-Inactivated</td>
		</tr>
		<tr>
			<td>FIJI (Fiji is Just Image J)</td>
			<td>NIH</td>
			<td>Version 1.52i</td>
			<td></td>
		</tr>
		<tr>
			<td>Fine Tip Tweezers</td>
			<td>F.S.T</td>
			<td>11254-20</td>
			<td>Style #5</td>
		</tr>
		<tr>
			<td>Fluorescence light source</td>
			<td>Excelitas</td>
			<td>012-63000</td>
			<td>X-Cite 120Q</td>
		</tr>
		<tr>
			<td>GFAP antibody</td>
			<td>Cell Signaling</td>
			<td>3670S</td>
			<td>GFAP (GA5) mouse monoclonal antibody</td>
		</tr>
		<tr>
			<td>Glass Bottom Dishes</td>
			<td>Mattek corporation</td>
			<td>P35G-1.5-14-C</td>
			<td>35 mm Dish | No. 1.5 Coverslip | 14 mm Glass Diameter | Uncoated</td>
		</tr>
		<tr>
			<td>Graefe Forceps</td>
			<td>F.S.T</td>
			<td>11054-10</td>
			<td>Graefe Iris Forceps with curved tips</td>
		</tr>
		<tr>
			<td>Green fluorescent dye that stains acidic compartments (late endosomes and lysosomes)</td>
			<td>Life Technologies</td>
			<td>L7526</td>
			<td>LysoTracker Green DND 26. Pre-dissolved in DMSOS to a 1mM stock solution. Dilute to the final working concentration in the growth medium or buffer of choice.</td>
		</tr>
		<tr>
			<td>Hank&#39;s Balanced Salt Solution (10x)</td>
			<td>Gibco</td>
			<td>14065-056</td>
			<td>Magnesium and calcium free</td>
		</tr>
		<tr>
			<td>Imaging Media</td>
			<td>Life Technologies</td>
			<td>A14291DJ</td>
			<td>Live Cell Imaging Solution</td>
		</tr>
		<tr>
			<td>Inverted Confocal Microscope</td>
			<td>Zeiss</td>
			<td></td>
			<td>LSM 780</td>
		</tr>
		<tr>
			<td>KymoToolBox</td>
			<td></td>
			<td></td>
			<td>https://github.com/fabricecordelieres/IJ_KymoToolBox</td>
		</tr>
		<tr>
			<td>Lipofection Enhancer Reagent</td>
			<td>Life Technologies</td>
			<td>11514015</td>
			<td>Plus Reagent</td>
		</tr>
		<tr>
			<td>Lipofection Reagent</td>
			<td>Life Technologies</td>
			<td>15338100</td>
			<td>Lipofectamine LTX reagent</td>
		</tr>
		<tr>
			<td>Orbital shaking incubator</td>
			<td>New Brunswick Scientific</td>
			<td>8261-30-1008</td>
			<td>Innova 4230 , orbital shaking incubator with temperature and speed control</td>
		</tr>
		<tr>
			<td>Penicillin/Strepomycin solution (100x)</td>
			<td>Gen Clone</td>
			<td>25-512</td>
			<td></td>
		</tr>
		<tr>
			<td>Phosphate Buffered Saline (10x)</td>
			<td>Gen Clone</td>
			<td>25-507x</td>
			<td></td>
		</tr>
		<tr>
			<td>Poly-D-Lysine Hydrobromide</td>
			<td>Sigma</td>
			<td>P7405</td>
			<td>Dissolve in Tris buffer, pH 8.5, at 1mg/mL. Freeze for long term storage. Avoid cycles of freezing and thawing</td>
		</tr>
		<tr>
			<td>Reduced serum medium</td>
			<td>Gibco</td>
			<td>31985-062</td>
			<td>OPTI-MEM</td>
		</tr>
		<tr>
			<td>Tissue Culture Flasks</td>
			<td>Olympus Plastics</td>
			<td>25-209</td>
			<td>75 cm^2. 100% angled neck access, 0.22um hydrophobic vented filter cap</td>
		</tr>
		<tr>
			<td>Tissue culture incubator</td>
			<td>Thermo Scientific</td>
			<td>51030285</td>
			<td>HERAcell VIOS 160i, tissue culture incubator with temperature, humidity, and CO2 control</td>
		</tr>
		<tr>
			<td>Tris-Base</td>
			<td>Sigma</td>
			<td>T1503</td>
			<td>8.402 g dissolved to one liter in water with 4.829 g Tris HCl to make 0.1 M Tris buffer, pH 8.5</td>
		</tr>
		<tr>
			<td>Tris-HCl</td>
			<td>Sigma</td>
			<td>T3253</td>
			<td>4.829 g dissolved to one liter in water with 8.402 g Tris Base to make 0.1 M Tris buffer, pH 8.5</td>
		</tr>
		<tr>
			<td>Trypsin-EDTA (0.25%), phenol red</td>
			<td>Gibco</td>
			<td>25200072</td>
			<td></td>
		</tr>
		<tr>
			<td>Vacuum-Driven Filter Systems</td>
			<td>Olympus Plastics</td>
			<td>25-227</td>
			<td>500 ml, PES membrane, 0.22 &#181;m</td>
		</tr>
		<tr>
			<td>Vannas scissors straight</td>
			<td>Roboz</td>
			<td>RS-5620</td>
			<td></td>
		</tr>
	</tbody>
</table>

visualizing and analyzing intracellular transport of organelles and other cargos in astrocytes

星形细胞是成人大脑中最丰富的细胞类型之一，它们在多种功能中起着关键作用。作为大脑平衡的中心角色，星形细胞为神经元提供重要的代谢物和缓冲细胞外水、离子和谷氨酸。星形细胞是"三部分"突触的组成部分，在突触的形成、修剪、维护和调制中也至关重要。为了启用这些高度交互的功能，星形细胞通过包括细胞在内的大量专用膜蛋白，相互之间以及与其他胶质细胞、神经元、脑血管和细胞外环境进行通信粘附分子、水孢素、孔通道、神经递质输送机和间隙结分子。为了支持这种动态通量，星形细胞，像神经元一样，依赖于紧密协调和有效的细胞内传输。与神经元不同，细胞内贩运被广泛划定，星形细胞中基于微管的迁移研究较少。然而，细胞膜蛋白的外泌和内分细胞贩运和细胞内细胞内传输协调星形细胞的正常生物学，这些过程往往在疾病或对损伤的反应中受到影响。在这里，我们提出一个直接的协议，培养高品质的鼠群细胞，荧光标记星形蛋白和感兴趣的细胞器，并记录其细胞内传输动力学使用时移共聚焦显微镜。我们还演示了如何使用可用的图像分析软件（即 ImageJ/FIJI）插件从获取的电影中提取和量化相关的传输参数。

上文概述的建立原小鼠MD星形细胞的协议应产生可重复的高质量培养物。虽然培养物最初包含星形细胞、成纤维细胞和其他胶质细胞的组合，包括微胶质和寡核苷酸细胞（图1Bi，Biv;红色箭头），但将AraC添加到DIV5-DIV7之间的混合培养物中，将这些污染物细胞的增殖。结合AraC处理和基于晃动的纯化策略丰富了星形细胞培养物的纯度（图1...

Watch this Scientific Journal Video about Visualizing and Analyzing Intracellular Transport of Organelles and Other Cargos in Astrocytes at JoVE.com

Visualizing and Analyzing Intracellular Transport of Organelles and Other Cargos in Astrocytes

在这里，我们描述了一种体外活成像方法，用于可视化细胞器的细胞内传输和在小鼠星形细胞中贩运血浆膜蛋白。该协议还提出了一种图像分析方法，以确定货物运输路线和动力学。

可视化和分析星形细胞中细胞内器官和其他货物的细胞内传输

Astrocytes are among the most abundant cell types in the adult brain, where they play key roles in a multiplicity of functions. As a central player in ...

我们的协议可用于研究在受控的细胞外环境和轻度疾病状态下星形细胞器或蛋白质的动能和定位。与提供蛋白质和细胞器定位单一快照的MHA固定制剂不同，我们的实时成像技术可用于分析单个活星细胞的粒子动力学。转染种子星核细胞在转染后24至48小时进行成像的适当密度的第二天。稀释血清介质中的唇部分离试剂，以适当的实验浓度。在250微升的血清介质中稀释5微克高纯度DNA，并在管中加入5微升的唇部增强剂试剂。将唇部分液试剂与同等体积的DNA唇部切除增强剂混合，在室温下孵育溶液15分钟。在孵育结束时，从星细胞培养物中去除上经剂，并将100微升的转染混合物滴投到细胞中。在37摄氏度和5%的二氧化碳下6小时后，用适当体积的星细胞培养基基物取代转染复合物，将细胞返回细胞培养箱24至72小时。成像前30分钟，在200微升星细胞培养培养基中稀释合适的利索体标记探针，以1微摩尔的工作浓度，在37摄氏度下用探针对星细胞进行30分钟的标记。然后用温暖的星细胞培养介质洗一次细胞，用成像介质代替洗涤。在探针标记后，立即将星细胞培养容器放入显微镜舞台上的适当适配器，并使用 EBI 荧光光，定位表达荧光蛋白或探针的细胞。使用数码相机调整荧光样品照明以可视化所选单元格，并调整焦点并放大到单个单元格。然后使用缩放和确定焦点函数以每两秒一帧的频率获取单个 Z 堆栈延时系列，时间间隔介于 300 到 500 秒之间。将延时图像保存并导出为 AVI 或 TIFF 堆栈文件。要分析延时图像，请打开 ImageJ 或斐济中的延时图像序列，并使用拆分通道工具拆分通道。在八位绿色通道图像中，使用分段线工具，使用所有影片的相同所需方向约定，沿粒子轨迹跟踪一条线，以便极性在所有被研究的单元格内和跨所有单元格之间保持一致。双击线条工具以调整线条宽度以匹配轨道的厚度，并使用 10 的线宽运行 Kymo ToolBox 插件的绘制 Kymo 宏。将出现一个提示，要求校准图像的时间和空间。校准后，将生成基纳仪，并可保存为 TIFF 文件。要分配粒子轨迹，请使用分段线工具在采集整个持续时间内手动跟踪 kymograph 中的每个粒子，并使用 ROI 管理器将每个粒子轨迹记录为感兴趣的区域。保存每个延时视频的所有感兴趣区域，以进行进一步分析，并运行 Kymo ToolBox 插件的分析 Kymo 宏。窗口将打开，要求从下拉菜单中从感兴趣的细胞核定义粒子运动的外向。应根据软件对每个感兴趣的货物的灵敏度定义限速，并且应调整线宽以匹配每个粒子轨迹的厚度。日志所有数据和日志外推坐标应用于计算各种货物运输参数。单击"确定"。然后，每个轨道计算的数据将按 kymograph 进行池，并保存在特定于每个图像的文本文件中。将细胞素阿拉伯边治疗与基于摇动的纯化策略相结合，比仅包括纯化步骤的传统协议更丰富了星细胞培养物的纯度。基于唇裂的转染允许蛋白质的瞬态表达，其水平是活细胞成像的最佳水平，而不会引起毒性或影响星细胞的生存能力。同样，使用荧光探针可以快速有效地标记酸性内糖体细胞器，以跟踪细胞器动力学和星形细胞。成像的延时数据可用于生成跟踪货物在时间和空间上的运动的 kymgraph。在这些 kymograph 中，指示货物的逆极运动由带负斜面的轨迹表示，而逆行运动由带正斜面的轨迹表示。固定囊泡显示为垂直轨迹。在这项分析中，通过星细胞区域对特定货物的通量进行量化，揭示了货物之间模态粒子的百分比差异，这些粒子可能代表其在获得电影的星细胞区域的正常碱线运动性。从 kymgraph 获得的每个粒子的 X、Y 位置沿完整时间刻度的精确映射也可用于评估其他运动参数，如货物速度和运行长度。由于该协议要求高质量的星细胞培养和高效的货物标签，因此应调整每个质粒或感兴趣的货物转染试剂孵育时间和采集时间表。该协议可以修改，以描述星形细胞中因细胞损伤、毒性、致病突变、突触活动或细胞内或细胞外环境变化而发生的运输事件。

Research

JoVE Journal

Neuroscience

Fluorescence Recovery After Photobleaching (FRAP) of Fluorescence Tagged Proteins in Dendritic Spines of Cultured Hippocampal Neurons

Fluorescence Recovery After Photobleaching FRAP of Fluorescence Tagged Proteins in Dendritic Spines of Cultured Hippocampal Neurons

漂白后荧光标记蛋白的荧光恢复（FRAP）在培养海马神经元的树突棘

FRAP has been used to quantify the mobility of GFP-tagged proteins. Using a strong excitation laser, the fluorescence of a GFP-tagged protein is bleached ...

科研

神经科学

Isolation and Culture of Mouse Cortical Astrocytes

小鼠脑皮层星形胶质细胞的分离和培养

Astrocytes  are an abundant cell type in the mammalian brain, yet much remains to be  learned about their molecular and functional characteristics. In ...

Imaging Intracellular Ca2+ Signals in Striatal Astrocytes from Adult Mice Using Genetically-encoded Calcium Indicators

Imaging Intracellular Ca2+ Signals in Striatal Astrocytes from Adult Mice Using Genetically-encoded Calcium Indicators

成像细胞内Ca<sup&gt; 2+</sup&gt;从成年小鼠使用基因编码的钙指标纹状体星形胶质细胞信号

Astrocytes display spontaneous intracellular Ca2+ concentration fluctuations ([Ca2+]i) and in several settings respond to neuronal excitation with ...

Protocol for Three-dimensional Confocal Morphometric Analysis of Astrocytes

协议星形胶质细胞的三维共聚焦形态分析

As glial cells in the brain, astrocytes have diverse functional roles in the central nervous system. In the presence of harmful stimuli, astrocytes modify ...

Analyzing Dendritic Morphology in Columns and Layers

分析树突状形态的列和图层

In many regions of the central nervous systems, such as the fly optic lobes and the vertebrate cortex, synaptic circuits are organized in layers and ...

Visualization and Live Imaging of Oligodendrocyte Organelles in Organotypic Brain Slices Using Adeno-associated Virus and Confocal Microscopy

腺相关病毒和共聚焦显微镜对器官脑切片胶质细胞器的可视化和实时成像

Neurons rely on the electric insulation and trophic support of myelinating oligodendrocytes. Despite the importance of oligodendrocytes, the advanced ...

Analyzing the Size, Shape, and Directionality of Networks of Coupled Astrocytes

耦合星形胶质细胞网络的大小、形状和方向性分析

It has become increasingly clear that astrocytes modulate neuronal function not only at the synaptic and single-cell levels, but also at the network ...

Analyzing Mitochondrial Transport and Morphology in Human Induced Pluripotent Stem Cell-Derived Neurons in Hereditary Spastic Paraplegia

遗传性痉挛性截瘫中人类诱导多能干细胞衍生神经元的线粒体迁移与形态分析

Neurons have intense demands for high energy in order to support their functions. Impaired mitochondrial transport along axons has been observed in human ...

Axonal Transport of Organelles in Motor Neuron Cultures using Microfluidic Chambers System

使用微流体腔系统在运动神经元培养物中有机细胞的斧头迁移

Motor neurons (MNs) are highly polarized cells with very long axons. Axonal transport is a crucial mechanism for MN health, contributing to neuronal ...

In Vitro Wedge Slice Preparation for Mimicking In Vivo Neuronal Circuit Connectivity

体外楔形切片准备模拟在Vivvo神经元电路连接

In vitro slice electrophysiology techniques measure single-cell activity with precise electrical and temporal resolution. Brain slices must be relatively ...