Name: ex vivo imaging of cell-specific calcium signaling at the tripartite synapse of the mouse diaphragm
Uploaded: 2018-10-04T14:00:00
Description: Watch this Scientific Journal Video about Ex Vivo Imaging of Cell-specific Calcium Signaling at the Tripartite Synapse of the Mouse Diaphragm at JoVE.com

这项工作得到了来自国家卫生研究院 (NIH) GM103554 和 GM110767 (东华三院) 和来自国家研究资源5P20RR018751 和国立普通医学科学院 8P20 GM103513 (G.W.H.) 的资金的支持。

畜牧和实验是按照国家卫生研究院的指导, 为实验室动物的护理和使用和 IACUC 在内华达大学。
1. 转基因小鼠横膈膜和膈神经的制备
<ol><li>购买转基因小鼠和寡核苷酸引物对这些小鼠进行基因型。 注: 这些小鼠的 "信息" 页面上都列出了引物。<ol>
<li>饲养一只3至6月大的老鼠, 表达一份适当的转基因/敲入的GCaMP3/6等位基因的副本, 并与同一年龄的第二只老鼠表达条件 GCaM...

<ol>
	<li>Sanes, J. R., Lichtman, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+the+vertebrate+neuromuscular+junction.">Development of the vertebrate neuromuscular junction.</a> Annual Review of Neuroscience. 22, 389-442 (1999).</li><li>Darabid, H., Perez-Gonzalez, A. P., Robitaille, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuromuscular+synaptogenesis:+coordinating+partners+with+multiple+functions.">Neuromuscular synaptogenesis: coordinating partners with multiple functions.</a> Nature Reviews Neuroscience. 15 (11), 703-718 (2014).</li><li>Fatt, P., Katz, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+analysis+of+the+end-plate+potential+recorded+with+an+intracellular+electrode.">An analysis of the end-plate potential recorded with an intracellular electrode.</a> Journal of Physiology. 115 (3), 320-370 (1951).</li><li>Todd, K. J., Robitaille, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Purinergic+modulation+of+synaptic+signalling+at+the+neuromuscular+junction.">Purinergic modulation of synaptic signalling at the neuromuscular junction.</a> Pflugers Archive. 452 (5), 608-614 (2006).</li><li>Hennig, G. 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=Use+of+Genetically+Encoded+Calcium+Indicators+(GECIs)+Combined+with+Advanced+Motion+Tracking+Techniques+to+Examine+the+Behavior+of+Neurons+and+Glia+in+the+Enteric+Nervous+System+of+the+Intact+Murine+Colon.">Use of Genetically Encoded Calcium Indicators (GECIs) Combined with Advanced Motion Tracking Techniques to Examine the Behavior of Neurons and Glia in the Enteric Nervous System of the Intact Murine Colon.</a> Frontiers of Cellular Neuroscience. 9, 436 (2015).</li><li>Heredia, D. J., Schubert, D., Maligireddy, S., Hennig, G. W., Gould, T. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Novel+Striated+Muscle-Specific+Myosin-Blocking+Drug+for+the+Study+of+Neuromuscular+Physiology.">A Novel Striated Muscle-Specific Myosin-Blocking Drug for the Study of Neuromuscular Physiology.</a> Frontiers of Cellular Neuroscience. 10, 276 (2016).</li><li>Johnson, B. R., Hauptman, S. A., Bonow, R. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Construction+of+a+simple+suction+electrode+for+extracellular+recording+and+stimulation.">Construction of a simple suction electrode for extracellular recording and stimulation.</a> Journal of Undergraduate Neuroscience Education. 6 (1), A21-A26 (2007).</li><li>Hong, S. J., Chang, C. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+geographutoxin+II+(mu-conotoxin)+for+the+study+of+neuromuscular+transmission+in+mouse.">Use of geographutoxin II (mu-conotoxin) for the study of neuromuscular transmission in mouse.</a> British Journal of Pharmacology. 97 (3), 934-940 (1989).</li><li>Heredia, D. J., Feng, C. Y., Hennig, G. W., Renden, R. B., Gould, T. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Activity-induced+Ca2++signaling+in+perisynaptic+Schwann+cells+of+the+early+postnatal+mouse+is+mediated+by+P2Y1+receptors+and+regulates+muscle+fatigue.">Activity-induced Ca2+ signaling in perisynaptic Schwann cells of the early postnatal mouse is mediated by P2Y1 receptors and regulates muscle fatigue.</a> Elife. 7, e30839 (2018).</li><li>Cho, J. 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=The+GCaMP-R+Family+of+Genetically+Encoded+Ratiometric+Calcium+Indicators.">The GCaMP-R Family of Genetically Encoded Ratiometric Calcium Indicators.</a> ACS Chemical Biology. 12 (4), 1066-1074 (2017).</li></ol>

在这里, 我们提供了一些例子, 以测量 GECI 表达小鼠完整的神经肌肉组织中的特定细胞的 Ca2 +反应。为了成功地进行这些实验, 在解剖过程中不应损伤膈神经。要在雪旺细胞中以低功率或大功率 (即20X 或 60X) 图像 Ca2 +响应, 必须使用 BHC 或µ-芋螺毒素来阻止移动。对于在肌肉细胞中的 Ca2 +反应的低功率成像, 有可能在没有这些药物的情况下测量它们, 从而允许同时获得?...

NMJ 像所有的突触一样, 由三元素组成: 一个突触前终端, 从神经元, 突触后神经元/效应细胞, perisynaptic 胶质细胞1,2。虽然突触传输的基本方面在这个突触3首先被证明, 这个过程的许多方面仍然不明, 部分由于这个突触的不同的细胞元素表达相同的分子。例如, 由脊椎动物 NMJ 的运动神经元共同释放的嘌呤腺嘌呤核苷酸 ATP 和乙酰胆碱的受体, 由肌肉、雪旺细胞和运动神经元表达, 从而使任何这些物质施加的功能效应 (例如, 发射机释放或反应, 肌肉力量世代)4。此外, 虽然 NMJ 的三方成分比较简单, 例如, 中枢神经系统中的神经元通常呈现多个突触输入, 无论是运动神经元、肌细胞还是雪旺细胞因刺激而变化关于它们的内在异质性 (如胚胎衍生、纤维亚型、形态学) 尚不清楚。为了解决这些问题, 在一个突触元素中同时跟踪许多细胞的反应, 以及在其他任何一个单独的元素中同时跟踪这种反应是有利的。使用化学染料来测量钙信号的常规策略不能达到这两个目标, 因为沐浴应用染料在应用到组织后被多种细胞类型所占, intracellularly 载染料只能用于可视化单个或小的细胞群。在这里, 利用转基因小鼠表达 GECIs 设计来测量细胞特定的钙信号, 连同具体的成像和软件工具5, 我们展示了这两个总体目标的第一个, 并讨论如何添加新的转基因工具将有助于实现第二个目标。这项技术将是有用的任何人感兴趣的跟踪钙动力学或其他细胞信号事件可观测到通过基因编码的光学传感器在多个细胞群体的同时。

<table>
	<tbody>
		<tr>
			<td>Myf5-Cre mice</td>
			<td>Jax</td>
			<td>#007893</td>
			<td>Drives muscle cell expression as early as E136</td>
		</tr>
		<tr>
			<td>Wnt1-Cre mice</td>
			<td>Jax</td>
			<td>#003829</td>
			<td>Drives expression into all&#160; Schwann cells at E13 but not P209</td>
		</tr>
		<tr>
			<td>Sox10-Cre mice</td>
			<td>Jax</td>
			<td>#025807</td>
			<td>Drives Schwann cell expression at older ages</td>
		</tr>
		<tr>
			<td>Conditional GCaMP3 mice</td>
			<td>Jax</td>
			<td>#029043</td>
			<td>Expresses GCaMP3 in cell-specific fashion</td>
		</tr>
		<tr>
			<td>Conditional GCaMP6f mice</td>
			<td>Jax</td>
			<td>#024105</td>
			<td>Expresses GCaMP6f in cell-specific fashion</td>
		</tr>
		<tr>
			<td>BHC (3-(N-butylethanimidoyl)-4-hydroxy-2H-chromen-2-one)</td>
			<td>Hit2Lead</td>
			<td>#5102862</td>
			<td>Blocks skeletal muscle myosin but not neurotransmission6</td>
		</tr>
		<tr>
			<td>CF594-&#945;-BTX</td>
			<td>Biotium</td>
			<td>#00007</td>
			<td>Labels acetylcholine receptor clusters at NMJ</td>
		</tr>
		<tr>
			<td>&#181;-conotoxin GIIIb</td>
			<td>Peptides Int&#39;l</td>
			<td>#CONO20-01000</td>
			<td>Blocks Nav1.4 voltage-dependent sodium channel8</td>
		</tr>
		<tr>
			<td>Silicone Dielectric Gel; aka Sylgard</td>
			<td>Ellswoth Adhesives</td>
			<td># Sil Dielec Gel .9KG&#160;</td>
			<td>Allows for the immobilization of the diaphragm by minutien pins</td>
		</tr>
		<tr>
			<td>Minutien pins (0.1mm diameter)</td>
			<td>Fine Science Tools</td>
			<td>26002-10</td>
			<td>Immobilizes diaphragm onto silicone dielectric gel</td>
		</tr>
		<tr>
			<td>Eclipse FN1 upright microscope&#160;</td>
			<td>Nikon</td>
			<td>MBA74100</td>
			<td>Allows staging and observation of specimen</td>
		</tr>
		<tr>
			<td>Basic Fixed Microscope Platform with Manual XY Microscope Translator&#160;</td>
			<td>Autom8</td>
			<td>MXMScr</td>
			<td>Allows movement of specimen</td>
		</tr>
		<tr>
			<td>Manual micromanipulator&#160;</td>
			<td>Narishige</td>
			<td>M-152</td>
			<td>Holds recording and stimulating electrodes&#160;</td>
		</tr>
		<tr>
			<td>Microelectrode amplifier&#160;</td>
			<td>Molecular Devices</td>
			<td>Axoclamp 900A</td>
			<td>Allows sharp electrode intracellular electrophysiological recording</td>
		</tr>
		<tr>
			<td>Microelectrode low-noise data acquisition system</td>
			<td>Molecular Devices</td>
			<td>Digidata 1550&#160;</td>
			<td>Allows electrophysiological data acquisition</td>
		</tr>
		<tr>
			<td>Microelectrode data analysis system</td>
			<td>Molecular Devices</td>
			<td>PCLAMP 10 Standard</td>
			<td>Performs electrophysiological data analysis</td>
		</tr>
		<tr>
			<td>Square wave stimulator</td>
			<td>Grass</td>
			<td>S48</td>
			<td>Stimulates nerve to excite muscle</td>
		</tr>
		<tr>
			<td>Stimulus Isolation Unit</td>
			<td>Grass</td>
			<td>PSIU6</td>
			<td>Reduces&#160; stimulation artifacts</td>
		</tr>
		<tr>
			<td>Borosilicate filaments, 1.0 mm outer diameter, 0.5mm internal diameter&#160;</td>
			<td>Sutter</td>
			<td>FG-GBF100-50-15</td>
			<td>Impales and records nerve-evoked muscle potentials</td>
		</tr>
		<tr>
			<td>Borosilicate filaments, 1.5 mm outer diameter, 1.17mm internal diameter&#160;</td>
			<td>Sutter</td>
			<td>BF150-117-15</td>
			<td>Lengthened and used for suction electrode</td>
		</tr>
		<tr>
			<td>Micropipette Puller</td>
			<td>Sutter</td>
			<td>P-97&#160;</td>
			<td>Pulls and prepares recording electrodes</td>
		</tr>
		<tr>
			<td>1200x1200 pixel, back-illuminated cMOS camera&#160;</td>
			<td>Photometrics</td>
			<td>Prime 95b</td>
			<td>Sensitive camera that allows high-resolution, high-speed imaging</td>
		</tr>
		<tr>
			<td>Light Source</td>
			<td>Lumencor</td>
			<td>Spectra X</td>
			<td>Provides illumination from LEDs for fluorescence obsevation</td>
		</tr>
		<tr>
			<td>&#160;Infinity-corrected fluorescent water immersion objectives, W.D. 2mm&#160;</td>
			<td>Nikon</td>
			<td>CFI60</td>
			<td>Provide long working distances for visualization of specimen</td>
		</tr>
		<tr>
			<td>Fiber Optic Illuminator with Halogen lamp</td>
			<td>Sumita</td>
			<td>LS-DWL-N</td>
			<td>Provides illumination for brightfield observation</td>
		</tr>
		<tr>
			<td>W-View Gemini Image Splitter&#160;</td>
			<td>Hamamatsu</td>
			<td>A12801-01</td>
			<td>Projects 1 pair of dual wavelength images separated by a dichroic to single camera</td>
		</tr>
		<tr>
			<td>Single-band Bandpass Filters&#160; (512/25-25 and 630/92-25)&#160;</td>
			<td>SemRock</td>
			<td>FF01-512/25-25; FF01-630/92-25</td>
			<td>Permits dual band imaging</td>
		</tr>
		<tr>
			<td>560 nm Single-Edge Dichroic Beamsplitter</td>
			<td>Sem Rock</td>
			<td>FF560-FDi01-25x36</td>
			<td>Dichroic mirror which separates beams of light to allow dual-wavelength imaging</td>
		</tr>
		<tr>
			<td>Imaging data acquisition system</td>
			<td>Nikon</td>
			<td>NIS Elements - MQS31000</td>
			<td>Allows imaging data acquisition</td>
		</tr>
		<tr>
			<td>Wavelength control module</td>
			<td>Nikon</td>
			<td>MQS41220</td>
			<td>Module for imaging data acqusiition</td>
		</tr>
		<tr>
			<td>Emission splitter hardware module&#160;</td>
			<td>Nikon</td>
			<td>MQS41410</td>
			<td>Module for imaging data acqusiition</td>
		</tr>
		<tr>
			<td>Imaging data analysis system</td>
			<td>NA</td>
			<td>Volumetry 8D5, Fiji</td>
			<td>Allows analysis of fluorescence intensity and other imaging data</td>
		</tr>
	</tbody>
</table>

ex vivo imaging of cell-specific calcium signaling at the tripartite synapse of the mouse diaphragm

细胞在组织中的电活动可以通过电生理技术进行监测, 但通常仅限于对单个细胞的分析。由于细胞质中的细胞内钙 (Ca2 +) 的增加往往是由于电活动, 或对无数其他刺激的反应, 这一过程可以监测的影像, 加载与荧光钙敏感染料。 然而, 由于这些染料被组织内的所有细胞类型占去, 很难在整个组织内的单个细胞类型中想象这种反应。相比之下, 基因编码的钙指标 (GECIs) 可以用单个细胞类型和荧光来表达, 以响应胞内 ca2 +的增加, 从而允许在整个人群中的 ca2 +信号的成像单个单元格类型。在这里, 我们应用 GECIs GCaMP3/6 的小鼠神经肌肉连接, 一个三方突触之间的运动神经元, 骨骼肌, 和终端/perisynaptic 雪旺细胞。我们展示了这种技术在经典的活体组织制剂中的效用。使用光学分光器, 我们执行动态 Ca2 +信号的双波长成像和神经肌肉连接的静态标签 (NMJ) 的方法, 可以很容易地适应监测两个细胞特定的 GECI 或基因编码的电压指标 (GEVI) 同时进行。最后, 我们讨论了用于捕捉荧光强度空间图的例程。这些光学、转基因和分析技术可以用来研究 NMJ 中不同细胞亚群在多种环境下的生物活性。

一些荧光强度变化的例子, 介导的增加细胞内钙2 +在定义的细胞类型的 NMJ, 显示了这种方法的效用。这些结果显示为空间荧光强度图, 它提供了响应细胞的位置, 以及它们的响应强度, 从而使人们能够评估多少细胞反应, 以及每个细胞对特定刺激。例如, 如图 1所示, 我们在 P7 Wnt1-Cre的膜片 NMJs 上拍摄了一个终端/perisynaptic 许旺细胞 (TPSCs) ?...

Watch this Scientific Journal Video about Ex Vivo Imaging of Cell-specific Calcium Signaling at the Tripartite Synapse of the Mouse Diaphragm at JoVE.com

Ex Vivo Imaging of Cell-specific Calcium Signaling at the Tripartite Synapse of the Mouse Diaphragm

在这里, 我们提出了一个协议, 以图像钙信号在个别细胞类型的人群在小鼠神经肌肉连接。

前体小鼠膈肌三方突触细胞特异性钙信号的影像学研究

The electrical activity of cells in tissues can be monitored by electrophysiological techniques, but these are usually limited to the analysis of ...

这种方法可以帮助回答神经肌肉领域的关键问题，如钙信号在肌肉或Schwann细胞中对神经刺激的反应的作用。这种技术的主要优点是，可以形象地激活特定的细胞类型，以回应神经刺激。首先，获得转基因小鼠，并基因型它们，如文本协议中详细说明。安乐死后，用虹细胞切除术剪刀横切整个动物，在肝脏下方和心脏和肺部上方。解剖肝脏、心脏和肺部。小心保持一个足够长的咽神经长度，以被吸引到吸力电极中。进一步去除肋骨笼和椎柱，隔膜周围的薄脊除外。将隔膜和肾神经样本放在微模糊管中，用 Krebs-Ringer 溶液，每毫升含有一微克的荧光标记的α邦加罗毒素。让试样在黑暗中在溶液中孵育10分钟。使用小针固定隔膜，将其固定在涂有硅胶的六厘米盘上，并填充约八毫升氧氧克雷布斯-林格溶液。将盘子放在显微镜的舞台上。然后，以每分钟 8 毫升的速度将隔膜与更多 Krebs-Ringer 溶液相近，使用 30 分钟。在四倍放大时，使用微操纵器将吸力电极移动到左神经上。通过拉出连接到吸管的五毫升注射器筒，施加吸力。通过使用 Brightfield 照明目视检查隔膜，确保隔膜收缩以响应一赫兹刺激。如果没有，通过增量转动电压旋钮来调节电压，从而达到一个超能量脉冲，可以通过肌肉收缩的目视检查来验证。如果仍然不可见，用注射器吹出神经，并尝试通过吸力再次吸入。现在，关闭灌注添加肌肉特异性肌素抑制剂BHC或电压门控钠通道拮抗性木康多毒素。在一毫升的克雷布斯-林格溶液中，将四根200毫摩尔库存BHC的微升移液，准备BHC预稀释。从盘子中取出一毫升的克雷布斯-林格溶液。然后将预稀释的BHC加入到菜中。30 分钟后，再打开新鲜 Krebs-Ringer 溶液的灌注 20 到 30 分钟。戴上手套，将外径为一毫米、内径为 0.4 毫米的硅丝玻璃放入微移子拉拔器中，准备记录电极。拧紧表盘以将其夹紧到位并关闭拉拔门。然后对拉拔器进行编程，如文本协议中所述。对于胚胎隔膜，使用放大器的软件控制测量电阻。确保电阻接近 60 兆欧姆，对于较旧的隔膜，10 到 20 兆欧姆。现在，用三个摩尔氯化钾加载记录电极。在放大倍数 10 倍时，使用舞台对面的第二个微操纵器将电极降为肌肉，作为刺激电极。使用电生理数据采集软件等待，直到静膜电位从零到零 65 毫伏以下变化。刺激在一赫兹，并验证肌肉动作潜力的存在，通过检查一个大的潜力，表现出一个适度的过冲。不要将刺激工件与动作电位混淆。在20倍的放大率下，通过寻找在绿色黄光激发下荧光标记的α bungarotoxin神经肌肉结，定位肌肉中心的端板带。切换到蓝光激发，以在肌肉、运动神经元或施万细胞中成像钙反应。在此示例中，GCAMP 3 以肌肉细胞表示。如果需要，使用带通滤波器和双波长成像的双色单刃滤波器设置图像分割器。使用查找表栏上的亮度条进行实验，将亮度条设置为表示组织的遗传编码钙指示器在 20 倍放大时表现出饱和度，而不会对氯化钾进行装箱。以每秒 20 帧的速度录制，因此不会错过任何快速事件。使用吸电极提供一系列脉冲，并收集一个细胞子类型的动态荧光钙反应以及静态荧光标记的α bungarotoxin神经肌肉结信号，以 1 至 45 秒的 20 至 40 赫兹刺激神经刺激。药理激动剂也可以通过沐浴应用或灌注添加，动态荧光钙反应可以收集相同的方式。当成像或电生理实验完成，因为已经取得了预期的结果，通过灌注线吸水，并通过吸水电极吸水两到三次，以确保盐不积聚。Schwann 细胞钙对 Wnt 一个 GCaMP 三小鼠隔膜在产后第 7 天的肾神经刺激的空间图显示了对神经肌肉结端寄生 Schwann 细胞的限制。同样的隔膜在贴有荧光标记的α双糖毒素后被成像，该标记键连接，使神经肌肉结的乙酰胆碱受体。同样的隔膜也在光明场照明下被成像，以引导记录电极到神经肌肉结的肌肉突触后区域。此处显示的是颜色编码细胞的空间划分或用于分析单个钙瞬变的感兴趣区域。在肌素抑制剂BHC存在的情况下，第5个月第5个GCaMP三只小鼠膜片的肌肉细胞钙反应的空间图显示了整个区域的所有膜片肌肉细胞的反应。相比之下，在粘膜毒素存在的情况下刺激同一膜片会导致所有与乙酰胆碱受体聚类富集端板带对应的膜片肌肉细胞的中区空间受限钙反应。按照这个程序，其他方法，如免疫组织化学或免疫印迹可以执行，以回答其他问题，如，什么是细胞的分子配置文件由电生理学或钙成像？这种方法可以深入了解Schwann细胞和肌肉细胞对正常新生儿小鼠神经刺激的种群反应。它也可以应用于研究老年小鼠或研究小鼠表达神经肌肉疾病相关蛋白质。

Research

JoVE Journal

Neuroscience

Ex Vivo Imaging of Cell-specific Calcium Signaling at the Tripartite Synapse of the Mouse Diaphragm

Ex Vivo Calcium Imaging for Visualizing Brain Responses to Endocrine Signaling in Drosophila

Ex Vivo Calcium Imaging for Visualizing Brain Responses to Endocrine Signaling in Drosophila

前体钙成像用于可视化大脑对果蝇内分泌信号的反应

Organ-to-organ communication by endocrine signaling, for example, from the periphery to the brain, is essential for maintaining homeostasis. As a model ...

科研

神经科学

In vivo Calcium Imaging of Mouse Geniculate Ganglion Neuron Responses to Taste Stimuli

在活体钙成像小鼠致病性甘里昂神经元对味觉刺激的反应

Within the last ten years, advances in genetically encoded calcium indicators (GECIs) have promoted a revolution in in vivo functional imaging. Using ...

In vivo Calcium Imaging in Mouse Inferior Olive

In vivo Calcium Imaging in Mouse Inferior Olive

在体内 小鼠劣质橄榄中的钙成像

Inferior olive (IO), a nucleus in the ventral medulla, is the only source of climbing fibers that form one of the two input pathways entering the ...

In Vivo Wide-Field and Two-Photon Calcium Imaging from a Mouse Using a Large Cranial Window

In Vivo Wide-Field and Two-Photon Calcium Imaging from a Mouse Using a Large Cranial Window

体内 使用大颅窗的小鼠宽场和双光子钙成像

Wide-field calcium imaging from the mouse&#39;s neocortex allows one to observe cortex-wide neural activity related to various brain functions. On the ...

Two-Photon and Wide-Field Calcium Imaging in the Superior Colliculus

Calcium Imaging in Mouse Superior Colliculus

Unveiling Neural Coding and Mechanisms of Visual Processing in the Superior Colliculus (Video) | JoVE

小鼠上丘的钙成像

The superior colliculus (SC), an evolutionarily conserved midbrain structure in all vertebrates, is the most sophisticated visual center before the ...

Mouse Retina OCT Fibergram Alignment with Confocal Images

Alignment of Visible-Light Optical Coherence Tomography Fibergrams with Confocal Images of the Same Mouse Retina

Advancements in In Vivo and Ex Vivo Retinal Imaging for Improved Glaucoma Diagnosis and Treatment (Video) | JoVE

可见光光学相干断层扫描光纤图与同一小鼠视网膜的共聚焦图像的对齐

In recent years, in vivo retinal imaging, which provides non-invasive, real-time, and longitudinal information about biological systems and processes, has ...

Evaluating Calcium Activity in Retinal Ganglion Cells Layer

Calcium Imaging In Electrically Stimulated Flat-Mounted Retinas

An Innovative Approach to Neural Electrical Stimulation Using Calcium Imaging (Video) | JoVE

电刺激平装视网膜中的钙成像

Retinal dystrophies are a leading cause of blindness worldwide. Extensive efforts are underway to develop advanced retinal prostheses that can bypass the ...

Monitoring Epileptiform Events in Drosophila via Calcium Imaging

Ex Vivo Calcium Imaging for Drosophila Model of Epilepsy

Insights into the Techniques and Findings of Recent Advancements in Epilepsy Research (Video) | JoVE

离体 癫痫 果蝇 模型的钙成像

Epilepsy is a neurological disorder characterized by recurrent seizures, partially correlated with genetic origin, affecting over 70 million individuals ...