Name: a fibrin-enriched and tpa-sensitive photothrombotic stroke model
Uploaded: 2021-06-04T12:30:00
Description: Watch this Scientific Journal Video about A Fibrin-Enriched and tPA-Sensitive Photothrombotic Stroke Model at JoVE.com

这项工作得到了美国国立卫生研究院（NIH）的资助（NS108763、NS100419、NS095064和HD080429给CYK和NS106592给YYS）。

该协议由弗吉尼亚大学机构动物护理和使用委员会 （IACUC） 批准，并遵循美国国立卫生研究院实验动物护理和使用指南。 图 1A 概述了该协议的外科手术顺序。
1. 手术设置
<ol><li>在手术前至少 15 分钟将温度设置为 37 °C 的加热垫放在小动物适配器上。为适配器准备一个鼻夹卷，允许动物头部旋转。准备麻醉剂氯胺酮 （60 mg/kg）/甲苯噻嗪 （10 mg/kg）。</...

<ol>
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Y., 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=Sickle+mice+are+sensitive+to+hypoxia/ischemia-induced+stroke+but+respond+to+tissue-type+plasminogen+activator+treatment.">Sickle mice are sensitive to hypoxia/ischemia-induced stroke but respond to tissue-type plasminogen activator treatment.</a> Stroke. 48 (12), 3347-3355 (2017).</li><li>Sun, Y. Y., Kuan, C. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+thrombotic+stroke+model+based+on+transient+cerebral+hypoxia-ischemia.">A thrombotic stroke model based on transient cerebral hypoxia-ischemia.</a> Journal of Visualized Experiments. (102), e52978 (2015).</li><li>Pena-Martinez, C., 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=Pharmacological+modulation+of+neutrophil+extracellular+traps+reverses+thrombotic+stroke+tPA+(tissue-type+plasminogen+activator)+resistance.">Pharmacological modulation of neutrophil extracellular traps reverses thrombotic stroke tPA (tissue-type plasminogen activator) resistance.</a> Stroke. 50 (11), 3228-3237 (2019).</li><li>Denorme, F., 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=ADAMTS13-mediated+thrombolysis+of+t-PA-resistant+occlusions+in+ischemic+stroke+in+mice.">ADAMTS13-mediated thrombolysis of t-PA-resistant occlusions in ischemic stroke in mice.</a> Blood. 127 (19), 2337-2345 (2016).</li><li>Marder, V. J., 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+thrombi+retrieved+from+cerebral+arteries+of+patients+with+acute+ischemic+stroke.">Analysis of thrombi retrieved from cerebral arteries of patients with acute ischemic stroke.</a> Stroke. 37 (8), 2086-2093 (2006).</li><li>Bacigaluppi, M., Semerano, A., Gullotta, G. S., Strambo, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Insights+from+thrombi+retrieved+in+stroke+due+to+large+vessel+occlusion.">Insights from thrombi retrieved in stroke due to large vessel occlusion.</a> Journal of Cerebral Blood Flow and Metabolism. 39 (8), 1433-1451 (2019).</li><li>Sun, Y. Y., 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=A+murine+photothrombotic+stroke+model+with+an+increased+fibrin+content+and+improved+responses+to+tPA-lytic+treatment.">A murine photothrombotic stroke model with an increased fibrin content and improved responses to tPA-lytic treatment.</a> Blood Advances. 4 (7), 1222-1231 (2020).</li><li>Su, E. J., 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=Activation+of+PDGF-CC+by+tissue+plasminogen+activator+impairs+blood-brain+barrier+integrity+during+ischemic+stroke.">Activation of PDGF-CC by tissue plasminogen activator impairs blood-brain barrier integrity during ischemic stroke.</a> Nature Medicine. 14 (7), 731-737 (2008).</li><li>Gupta, A. 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1985 年引入的传统 RB 光血栓性卒中是一种有吸引力的局灶性脑缺血模型，适用于简单的外科手术、低死亡率和高可重复性脑梗死。5 在该模型中，光动力染料 RB 在光激发下迅速激活血小板，导致致密的聚集体阻塞血管 5,8,23。然而，RB诱导的血凝块中的少量纤维蛋白（图2）偏离了缺血性?...

血管内血栓切除术和 tPA 介导的溶栓是美国食品和药物管理局 （FDA） 批准的仅有的两种急性缺血性卒中疗法，美国每年有 ~700,000 名患者患病1。由于血栓切除术的应用仅限于大血管闭塞 （LVO），而 tPA 溶栓术可以缓解小血管闭塞，因此两者都是急性缺血性卒中2 的有价值的治疗方法。此外，两种疗法的联合使用（例如，在卒中发作后 4.5 小时内开始 tPA 溶栓，然后进行血栓切除术）可改善再灌注和功能结局3。因此，即使在血栓切除术时代，优化溶栓仍然是卒中研究的重要目标。
血栓栓塞模型是临床前卒中研究的重要工具，旨在改善溶栓治疗。这是因为机械血管闭塞模型（例如，腔内缝合 MCA 闭塞）不会产生血凝块，并且其在去除机械闭塞后脑血流的快速恢复被过度理想化 4,5。迄今为止，主要的血栓栓塞模型包括光血栓形成 6,7,8、局部氯化铁 （FeCl 3） 应用9、凝血酶显微注射到 MCA 分支 10,11、体外（微）栓子注射到 MCA 或颈总动脉 （CCA） 12、13、14 和短暂性缺氧缺血 （tHI）15,16，17,18.这些卒中模型在随后血栓的组织学组成和对tPA介导的裂解疗法的敏感性方面有所不同（表1）。它们在开颅手术要求（原位凝血酶注射和局部应用 FeCl3 需要）、梗死大小和位置的一致性（例如，CCA 输注微栓子产生非常可变的结果）以及对心血管系统的总体影响（例如，tHI 增加心率和心输出量以补偿缺氧诱导的外周血管舒张）方面也有所不同。
基于 RB 染料的光血栓卒中 （PTS） 模型具有许多吸引人的特征，包括简单的无开颅手术、低死亡率（通常< 5%）以及可预测的梗死大小和位置（在 MCA 供应区域），但它有两个主要局限性。8 首先需要注意的是，对 tPA 介导的溶栓治疗反应弱至零，这也是 FeCl3 模型7、19、20 的缺点。PTS 和 FeCl3 卒中模型的第二个警告是，随后的血栓由密集的血小板聚集体和少量纤维蛋白组成，这不仅导致其对 tPA 溶解治疗的弹性，而且还偏离了急性缺血性卒中患者中血小板：纤维蛋白血栓混合的模式21,22。相比之下，原位凝血酶显微注射模型主要包括聚合纤维蛋白和不确定含量的血小板10。
鉴于上述推理，我们假设 RB 和亚血栓形成剂量的凝血酶的混合物通过变薄的颅骨进行 MCA 靶向光活化可能会增加所得血栓中的纤维蛋白成分并提高对 tPA 介导的裂解治疗的敏感性。我们已经证实了这一假设，23 在此我们描述了改良 （T+RB） 光血栓中风模型的详细程序。

<table>
	<tbody>
		<tr>
			<td>2,3,5-triphenyltetrazolium chloride (TTC)</td>
			<td>Sigma</td>
			<td>T8877</td>
			<td>infarct</td>
		</tr>
		<tr>
			<td>4-0 Nylon monofilament suture</td>
			<td>LOOK</td>
			<td>766B</td>
			<td>surgical supplies</td>
		</tr>
		<tr>
			<td>5-0 silk suture</td>
			<td>Harvard Apparatus</td>
			<td>624143</td>
			<td>surgical supplies</td>
		</tr>
		<tr>
			<td>543nm laser beam</td>
			<td>Melles Griot</td>
			<td>25-LGP-193-249</td>
			<td>photothrombosis</td>
		</tr>
		<tr>
			<td>adult male mice</td>
			<td>Charles River</td>
			<td>C57BL/6</td>
			<td>10~14 weeks old (22~30 g)</td>
		</tr>
		<tr>
			<td>Anesthesia bar for mouse adaptor</td>
			<td>machine shop, UVA</td>
			<td></td>
			<td>surgical setup</td>
		</tr>
		<tr>
			<td>Avertin (2, 2, 2-Tribromoethanol)</td>
			<td>Sigma</td>
			<td>T48402</td>
			<td>euthanasia</td>
		</tr>
		<tr>
			<td>Dental drill</td>
			<td>Dentamerica</td>
			<td>Rotex 782</td>
			<td>surgical setup</td>
		</tr>
		<tr>
			<td>Digital microscope</td>
			<td>Dino-Lite</td>
			<td>AM2111</td>
			<td>brain imaging</td>
		</tr>
		<tr>
			<td>Dissecting microscope</td>
			<td>Olympus</td>
			<td>SZ40</td>
			<td>surgical setup</td>
		</tr>
		<tr>
			<td>Fine curved forceps (serrated)</td>
			<td>FST</td>
			<td>11370-31</td>
			<td>surgical instrument</td>
		</tr>
		<tr>
			<td>Fine curved forceps (smooth)</td>
			<td>FST</td>
			<td>11373-12</td>
			<td>surgical instrument</td>
		</tr>
		<tr>
			<td>goat anti-rabbit Alexa Fluro 488</td>
			<td>Invitrogen</td>
			<td>A11008</td>
			<td>Immunohistochemistry</td>
		</tr>
		<tr>
			<td>Halsted-Mosquito hemostats</td>
			<td>FST</td>
			<td>13008-12</td>
			<td>surgical instrument</td>
		</tr>
		<tr>
			<td>Heat pump with warming pad</td>
			<td>Gaymar</td>
			<td>TP700</td>
			<td>surgical setup</td>
		</tr>
		<tr>
			<td>infusion pump</td>
			<td>KD Scientific</td>
			<td>200</td>
			<td>thrombolytic treatment</td>
		</tr>
		<tr>
			<td>Insulin syringe with 31G needle</td>
			<td>BD</td>
			<td>328291</td>
			<td>photothrombosis</td>
		</tr>
		<tr>
			<td>Ketamine</td>
			<td>CCM, UVA</td>
			<td></td>
			<td>anesthesia</td>
		</tr>
		<tr>
			<td>Laser protective google 532nm</td>
			<td>Thorlabs</td>
			<td>LG3</td>
			<td>photothrombosis</td>
		</tr>
		<tr>
			<td>Meloxicam SR</td>
			<td>CCM, UVA</td>
			<td></td>
			<td>NSAID analgesia</td>
		</tr>
		<tr>
			<td>micro needle holders</td>
			<td>FST</td>
			<td>12060-01</td>
			<td>surgical instrument</td>
		</tr>
		<tr>
			<td>micro scissors</td>
			<td>FST</td>
			<td>15000-03</td>
			<td>surgical instrument</td>
		</tr>
		<tr>
			<td>MoorFLPI-2 blood flow imager</td>
			<td>Moor</td>
			<td>780-nm laser source</td>
			<td>Laser Speckle Contrast Imaging</td>
		</tr>
		<tr>
			<td>Mouse adaptor</td>
			<td>RWD</td>
			<td>68014</td>
			<td>surgical setup</td>
		</tr>
		<tr>
			<td>Puralube Vet ointment</td>
			<td>Fisher</td>
			<td>NC0138063</td>
			<td>eye dryness prevention</td>
		</tr>
		<tr>
			<td>Retractor tips</td>
			<td>Kent Scientific</td>
			<td>Surgi-5014-2</td>
			<td>surgical setup</td>
		</tr>
		<tr>
			<td>Rose Bengal</td>
			<td>Sigma</td>
			<td>198250</td>
			<td>photothrombosis</td>
		</tr>
		<tr>
			<td>Thrombin</td>
			<td>Sigma</td>
			<td>T7513</td>
			<td>photothrombosis</td>
		</tr>
		<tr>
			<td>Tissue glue</td>
			<td>Abbott Laboratories</td>
			<td>NC9855218</td>
			<td>surgical supplies</td>
		</tr>
		<tr>
			<td>tPA</td>
			<td>Genetech</td>
			<td>Cathflo activase 2mg</td>
			<td>thrombolytic treatment</td>
		</tr>
		<tr>
			<td>Vibratome</td>
			<td>Stoelting</td>
			<td>51425</td>
			<td>TTC infacrt</td>
		</tr>
		<tr>
			<td>Xylazine</td>
			<td>CCM, UVA</td>
			<td></td>
			<td>anesthesia</td>
		</tr>
	</tbody>
</table>

a fibrin-enriched and tpa-sensitive photothrombotic stroke model

理想的血栓栓塞性卒中模型需要某些特性，包括相对简单的外科手术、死亡率低、梗死大小和位置一致、血小板：纤维蛋白混合血凝块的沉淀与患者相似，以及对纤溶治疗具有足够的敏感性。基于孟加拉玫瑰 （RB） 染料的光血栓中风模型满足前两个要求，但对 tPA 介导的裂解处理高度难治，这可能是由于其富含血小板但纤维蛋白贫乏的凝块组成。我们认为，RB染料（50mg / kg）和亚血栓形成剂量的凝血酶（80 U / kg）的组合用于针对大脑中动脉（MCA）近端分支的光活化，可能会产生富含纤维蛋白和tPA敏感的凝块。事实上，凝血酶和RB（T+RB）联合光血栓形成模型触发了混合血小板：纤维蛋白血凝块，如免疫染色和免疫印迹所示，并保持一致的梗死大小和位置以及低死亡率。此外，在光激活后 2 小时内静脉注射 tPA（阿替普酶，10 mg/kg）可显着减小 T+RB 光血栓形成的梗死面积。因此，凝血酶增强的光血栓中风模型可能是测试新型溶栓疗法的有用实验模型。

首先，我们比较了RB与T+RB光血栓形成诱导的血凝块中的纤维蛋白含量。光活化后2 h通过心内灌注固定剂处死小鼠，取出脑进行纵横面MCA分支免疫荧光染色。在RB光血栓形成中，MCA分支密集地填充了CD41+血小板和少量纤维蛋白（图2A，C）。相比之下，T+RB光血栓形成中的MCA分支被随机混合的血小板：纤维蛋白凝块阻塞（图2B，D，n<...

Watch this Scientific Journal Video about A Fibrin-Enriched and tPA-Sensitive Photothrombotic Stroke Model at JoVE.com

A Fibrin-Enriched and tPA-Sensitive Photothrombotic Stroke Model

传统的光血栓中风 （PTS） 模型主要诱导对组织纤溶酶原激活剂 （tPA） 溶解治疗具有高耐药性的致密血小板聚集体。本文通过共注射凝血酶和光敏染料进行光活化，引入了改良的小鼠PTS模型。凝血酶增强的 PTS 模型产生混合血小板：纤维蛋白凝块，并且对 tPA 溶栓高度敏感。

纤维蛋白富集和 tPA 敏感的光血栓形成卒中模型

An ideal thromboembolic stroke model requires certain properties, including relatively simple surgical procedures with low mortality, a consistent ...

我们希望创建一个中风模型，该模型产生与患者样本相似的血凝块，并对临床相关的溶栓治疗做出反应。因此，我们修改了经典的光血栓中风模型，并在光活化之前混合凝血酶和光动力染料孟加拉玫瑰。我们很高兴该手术确实会产生对tPA介导的溶栓高度敏感的血凝块。这种改良的光血栓中风模型使用非常简单的外科手术，死亡率低，产生了高度一致的梗死大小和位置。您还会产生纤维蛋白和血小板混合血凝块，与急性卒中患者的血凝块相似。因此，我们认为开发更有效的溶栓疗法对于临床前卒中研究非常有用。我们希望更多的人使用我们的模型。演示该程序的将是我们的研究助理教授Yu-Yo Sun博士。手术前30分钟，通过注射镇痛剂来准备小鼠。麻醉鼠标后，捏住脚趾以确保其完全麻醉。然后用脱毛膏去除左脖子和头部的毛发。将鼠标放在仰卧位的小动物适配器上，并用聚维酮碘和70%乙醇交替擦拭三次，对手术皮肤区域进行消毒。在解剖显微镜下，使用一把微型剪刀和直镊子在中线外侧约 0.2 厘米处做一个 0.5 厘米的左侧颈椎切口。接下来，使用一对细锯齿镊子，拉开软组织和筋膜，露出LCCA。然后使用一把细光滑的镊子，小心地将LCCA与迷走神经分开。使用 5-0 丝线缝合线在 LCCA 周围放置永久性双结缝合线。将鼠标翻转到俯卧位，然后将鼻夹滚动 15 度。然后用三滴Betadine和70%乙醇交替擦拭皮肤，对手术区域进行消毒。用一把微型剪刀和直镊子沿着左眼和耳朵在头皮上做一个0.8厘米的切口，露出颞肌。接下来，沿着左顶骨颞肌边缘做一个 0.5 厘米的切口。然后在颞肌上做第二个0.3厘米的垂直切口，并缩回肌肉，露出顶骨和鳞状骨的边缘。确保可视化额骨和顶骨之间的冠状缝合线的里程碑。接下来，通过应用无菌生理盐水润湿颅骨以显示左侧MCA，并用记号笔在鳞状骨上标记近端MCA分支。用气动牙钻轻轻地在这个标记区域周围画一个直径为一毫米的圆圈。然后将头骨变薄约0.2毫米深，而不接触硬脑膜下方。当残留一层非常薄的骨头时停止钻孔。接下来，根据小鼠的体重混合凝血酶和玫瑰孟加拉溶液，并用 31 号针将混合物缓慢注入眶后窦。在双眼上涂抹眼药膏以防止干燥。用 532 纳米激光将照明器施加在两英寸距离内的钻孔部位 20 分钟。通过激光投影护目镜可视化 MCA 近端分支的照明。20 分钟后，停止激光照明。在颅骨的顶骨上做一个直径为三毫米的颅窗，并在上面放置一个盖玻片。然后在 20 倍水浸物镜下定位远端 MCA。在成像前五分钟通过尾静脉注射 DyLight 488 偶联的抗 GP1b-β 抗体标记循环血小板。然后眼眶后注射凝血酶和玫瑰孟加拉溶液混合物。使用直径为 10 微米的激光束的 532 纳米激光系统光激活 MCA，并记录图像直至血栓形成。对于tPA给药，将麻醉的动物放在37摄氏度的温暖垫上。在选定的光激活后时间点用 45 摄氏度的温湿纱布包裹它的尾巴一分钟。接下来，通过尾静脉注射重组人tPA。推注 50%，然后使用输液泵在 30 分钟内输注剩余的 50%。为了监测脑血流，在头皮上做一个中线切口，露出头骨。用无菌生理盐水润湿颅骨，然后轻轻地将超声波凝胶涂抹在颅骨上，确保避免凝胶中出现毛发或气泡。在激光散斑对比成像仪下监测两个大脑半球的脑血流 10 分钟。免疫荧光标记显示，在基于孟加拉玫瑰染料的光血栓形成中，MCA分支密集地填充了CD41阳性血小板和少量纤维蛋白。相反，在凝血酶加孟加拉玫瑰光血栓形成中，MCA分支被随机混合的血小板和纤维蛋白凝块阻塞。免疫印迹分析显示，与单独使用孟加拉玫瑰相比，凝血酶加孟加拉玫瑰光血栓形成的同侧半球纤维蛋白沉积增加两倍以上。基于共聚焦显微镜的活体成像显示，即使在激光照射下，静脉注射凝血酶也未能诱导血小板聚集。在孟加拉玫瑰光血栓形成模型中，血小板形成均匀的凝块，但在凝血酶加孟加拉玫瑰模型中，血小板聚集不均匀，有多个微弱区域。通过激光散斑对比成像测量同一只小鼠在 tPA 治疗前和后 24 小时与载体治疗的 CBF，并归一化到对侧半球。在孟加拉玫瑰光血栓形成中，tPA治疗导致CBF恢复的趋势，特别是在缺血性边界区域，与载体治疗的小鼠相比。在凝血酶加孟加拉玫瑰光血栓形成中，tPA处理小鼠CBF的恢复更为突出，近端MCA分支常在24小时可见。在孟加拉玫瑰光血栓形成中，在载体处理和tPA处理的小鼠中检测到相似的梗死大小。相比之下，在凝血酶加孟加拉玫瑰光血栓形成中，与载体治疗的小鼠相比，tPA 裂解治疗在光激活后 0.5、1 或 2 小时显着减少了梗死，但在光激活后 6 小时内没有，凝血酶介导的纤维蛋白生成对于该过程中的血栓形成至关重要。该方法可用于研究凝块组成并进一步研究溶栓治疗。在此程序之后，可以应用其他溶栓治疗方法并确定后期疗效。添加凝血酶可以将凝块组成从纤维蛋白贫乏调整为纤维蛋白富集，使研究人员能够制定临床相关的治疗策略。

Research

JoVE Journal

Neuroscience

Retrograde Loading of Nerves, Tracts, and Spinal Roots with Fluorescent Dyes

逆行神经，大港，并用荧光染料的神经根装载

Retrograde labeling of neurons is a standard anatomical method1,2 that has also been used to load calcium and voltage-sensitive dyes into neurons3-6. ...

科研

神经科学

Progenitor-derived Oligodendrocyte Culture System from Human Fetal Brain

来自祖，人脑少突胶质细胞培养系统

Differentiation of human neural progenitors into neuronal and glial cell types offers a model to study and compare molecular regulation of neural cell ...

Deriving the Time Course of Glutamate Clearance with a Deconvolution Analysis of Astrocytic Transporter Currents

星形细胞转运电流派生谷氨酸间隙的时间进程与反卷积分析

The highest density of glutamate transporters in the brain is found in astrocytes. Glutamate transporters couple the movement of glutamate across the ...

Utilizing 3D Printing Technology to Merge MRI with Histology: A Protocol for Brain Sectioning

利用3D打印技术来合并MRI与组织学：一个大脑切片协议

Magnetic resonance imaging (MRI) allows for the delineation between normal and abnormal tissue on a macroscopic scale, sampling an entire tissue volume ...

Evaluation of Bioenergetic Function in Cerebral Vascular Endothelial Cells

在脑血管内皮细胞生物能量功能的评价

The integrity of the blood-brain-barrier (BBB) is critical to prevent brain injury. Cerebral vascular endothelial (CVE) cells are one of the cell types ...

Measuring Synaptic Vesicle Endocytosis in Cultured Hippocampal Neurons

海马神经元中突触囊泡吞的测量

During endocytosis, fused synaptic vesicles are retrieved at nerve terminals, allowing for vesicle recycling and thus the maintenance of synaptic ...

An In Vivo Assessment of Blood-Brain Barrier Disruption in a Rat Model of Ischemic Stroke

An In Vivo Assessment of Blood-Brain Barrier Disruption in a Rat Model of Ischemic Stroke

一种体内评价大鼠缺血性中风模型中血脑屏障的破坏

Ischemic stroke leads to vasogenic cerebral edema and subsequent primary brain injury, which is mediated through destruction of the blood-brain barrier ...

A Preclinical Model to Assess Brain Recovery After Acute Stroke in Rats

评估大鼠急性中风后脑恢复的临床前模型

The purpose of this study was to establish and validate an animal brain ischemia model in the recovery and sequela stages. A middle cerebral artery ...

Time-lapse Live Imaging and Quantification of Fast Dendritic Branch Dynamics in Developing Drosophila Neurons

Time-lapse Live Imaging and Quantification of Fast Dendritic Branch Dynamics in Developing Drosophila Neurons

发展果蝇神经元的快速丹德林分体动力学的延时实时成像和定量

Highly motile dendritic filopodia are widely present in neurons at early developmental stages. These exploratory dynamic branches sample the surrounding ...

Confocal Microscopy to Measure Three Modes of Fusion Pore Dynamics in Adrenal Chromaffin Cells

共聚焦显微镜测量肾上腺嗜铬细胞融合孔隙动力学的三种模式

Dynamic fusion pore opening and closure mediate exocytosis and endocytosis and determine their kinetics. Here, it is demonstrated in detail how confocal ...