Name: high-resolution three-dimensional imaging of the footpad vasculature in a murine hindlimb gangrene model
Uploaded: 2022-03-16T13:00:00
Description: Watch this Scientific Journal Video about High-Resolution Three-Dimensional Imaging of the Footpad Vasculature in a Murine Hindlimb Gangrene Model at JoVE.com

这项工作得到了美国国立卫生研究院对Z-J L和OC V的资助[R01HL149452和VITA（NHLBl-CSB-HV-2017-01-JS）]的支持。我们还感谢迈阿密大学医学院迈阿密治疗瘫痪项目的显微镜和成像设施提供对其图像分析和处理软件的访问。

协议中描述的所有动物实验均已获得迈阿密大学机构动物护理和使用委员会（IACUC）的批准。FVB小鼠，包括雄性和雌性，年龄在8-12周，用于研究。
1. L-NAME溶液的制备
<ol><li>在层流罩中的无菌条件下，通过将1gL-NAME粉末（见 材料表）与20mL无菌水溶解以制成50mg / mL溶液来制备L-NAME储备溶液。将储备溶液储存在-20°C的300-500μL等分试样中长达3个月。</li>
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<ol>
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H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Signal+processing+methods+for+quantitative+power+doppler+microvascular+angiography.">Signal processing methods for quantitative power doppler microvascular angiography.</a> Electronic Thesis and Dissertation Repository. , 3106 (2015).</li><li>Matic, M., Matic, A., Djuran, V., Gajinov, Z., Prcic, S., Golusin, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Frequency+of+peripheral+arterial+disease+in+patients+with+chronic+venous+insufficiency.">Frequency of peripheral arterial disease in patients with chronic venous insufficiency.</a> Iranian Red Crescent Medical Journal. 18 (1), 1-6 (2016).</li><li>Ammermann, 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=Concomitant+chronic+venous+insufficiency+in+patients+with+peripheral+artery+disease:+Insights+from+MR+angiography.">Concomitant chronic venous insufficiency in patients with peripheral artery disease: Insights from MR angiography.</a> European Radiology. 30 (7), 3908-3914 (2020).</li><li>Yang, 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=Cellular+and+molecular+mechanism+regulating+blood+flow+recovery+in+acute+versus+gradual+femoral+artery+occlusion+are+distinct+in+the+mouse.">Cellular and molecular mechanism regulating blood flow recovery in acute versus gradual femoral artery occlusion are distinct in the mouse.</a> Journal of Vascular Surgery. 48 (6), 1546-1558 (2008).</li><li>Padgett, M. E., McCord, T. J., McClung, J. M., Kontos, C. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methods+for+acute+and+subacute+murine+hindlimb+ischemia.">Methods for acute and subacute murine hindlimb ischemia.</a> Journal of Visualized Experiments. (112), e54166 (2016).</li><li>Nowak-Sliwinska, P., 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=Consensus+guidelines+for+the+use+and+interpretation+of+angiogenesis+assays.">Consensus guidelines for the use and interpretation of angiogenesis assays.</a> Angiogenesis. 21 (3), 425-432 (2018).</li><li>Greco, A., 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=Repeatability,+reproducibility+and+standardisation+of+a+laser+doppler+imaging+technique+for+the+evaluation+of+normal+mouse+hindlimb+perfusion.">Repeatability, reproducibility and standardisation of a laser doppler imaging technique for the evaluation of normal mouse hindlimb perfusion.</a> Sensors. 13 (1), 500-515 (2013).</li><li>Kochi, T., 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=Characterization+of+the+arterial+anatomy+of+the+murine+hindlimb:+Functional+role+in+the+design+and+understanding+of+ischemia+models.">Characterization of the arterial anatomy of the murine hindlimb: Functional role in the design and understanding of ischemia models.</a> PLoS ONE. 8 (12), 84047 (2013).</li><li>Hlushchuk, R., Haberth&#252;r, D., Djonov, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ex+vivo+microangioCT:+Advances+in+microvascular+imaging.">Ex vivo microangioCT: Advances in microvascular imaging.</a> Vascular Pharmacology. 112, 2-7 (2019).</li><li>Robertson, R. 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虽然小鼠后肢缺血是研究PAD和CLTI新生血管形成的最广泛使用的临床前模型，但缺血严重程度和恢复程度存在显着差异，具体取决于所使用的特定小鼠品系以及动脉破坏的部位，数量和方法。股动脉结扎和L-NAME的IP给药相结合可以可靠地诱导FVB小鼠的后肢坏疽11。相同的治疗导致C57BL / 6小鼠的后肢缺血而没有组织损失，而在BALB / c小鼠中，足部或腿部的自动截肢可以仅由股动脉结扎...

外周动脉疾病 （PAD） 的特征是动脉粥样硬化导致四肢血流量减少，影响美国 650 万人和全球 2 亿人1。PAD 患者的肢体功能和生活质量下降，而患有 CLTI（最严重的 PAD 形式）的患者截肢和死亡风险增加，5 年死亡率接近 50%2。在临床实践中，踝臂指数 （ABI） <0.9 的患者被认为患有 PAD，而与静息痛或组织丢失相关的 ABI <.4 的患者则认为患有 CLTI3。具有相似 ABI 的患者的症状因日活动、肌肉对缺血的耐受性、解剖学变异和侧支发育差异而异4。指和肢体坏疽是导致CLTI的所有血管闭塞性疾病的最严重表现。它是一种干燥的坏死形式，使软组织木乃伊化。除了动脉粥样硬化性 PAD 外，还可以在糖尿病患者、血管炎（如 Buerger 病和雷诺氏现象）或终末期肾病情况下的钙化反应患者中观察到该功能5，6。
已经开发了几种临床前模型来研究PAD / CLTI的发病机制并测试潜在治疗方法的疗效，其中最常见的仍然是小鼠后肢缺血。诱导小鼠后肢缺血通常是通过阻塞髂动脉或股动脉的血流来完成的，通过缝合结扎，电凝或其他收缩所需血管的方法7。这些技术大大减少了后肢的灌注，并刺激大腿和小腿肌肉的新生血管形成。然而，小鼠菌株对缺血性损伤的敏感性存在明显的应变性差异，部分原因是侧支分布的解剖学差异8，9。例如，C57BL / 6小鼠对后肢缺血相对耐药，显示肢体功能降低，但通常没有证据表明脚垫中有坏疽。另一方面，BALB / c小鼠从缺血中恢复的能力天生较差，通常在单独股动脉结扎后发生足部或小腿的自动截肢。这种对缺血的严重反应会缩小治疗窗口，并排除对肢体再灌注和功能进行纵向评估。有趣的是，位于小鼠7号染色体上的单个数量性状位点的遗传差异与C57BL / 6和BALB / c小鼠对组织坏死和肢体再灌注的这些差异易感性有关10。
与C57BL / 6和BALB / c菌株相比，FVB小鼠对单独股动脉结扎表现出中间但不一致的反应。一些动物以黑色缺血性指甲或木乃伊手指的形式出现脚垫坏疽，而另一些动物则没有任何明显的缺血迹象11。同时给予Nω-硝基-L-精氨酸甲酯盐酸盐（L-NAME），一种一氧化氮合酶（NOS）抑制剂12，可防止代偿性血管扩张机制，并进一步增加后肢组织中的氧化应激。结合股动脉结扎或凝血，这种方法在FVB小鼠中持续产生足垫组织损失，类似于CLTI的萎缩性变化，但很少进展为肢体自动截肢11。氧化应激是PAD / CLTI的标志之一，并通过内皮功能障碍和一氧化氮（NO）生物利用度降低来传播13，14。NO 是一种多能分子，通常对动脉和毛细血管血流、血小板粘附和聚集以及白细胞募集和活化发挥有益作用13。NOS水平的降低也被证明可以激活血管紧张素转换酶，从而诱导氧化应激并加速动脉粥样硬化的进展15。
一旦建立了后肢缺血的模型，还需要监测随后的肢体再灌注和任何潜在治疗的治疗效果。在提出的小鼠坏疽模型中，可以首先使用Faber评分来量化组织损失的程度，以评估足部的毛坯外观（0：正常，1-5：指甲脱落，其中评分代表受影响的指甲数量，6-10：手指萎缩，其中分数代表受影响的手指数量，11-12：部分和完全足部萎缩， 分别）9.然后通常使用LDPI进行后肢灌注的定量测量，LDPI依赖于激光和红细胞之间的多普勒相互作用来指示感兴趣区域的像素水平灌注（ROI）16。虽然这种技术是定量的，非侵入性的，并且是重复测量的理想选择，但它不能提供后肢脉管系统的颗粒解剖细节16。其他成像方式，如显微计算机断层扫描 （micro-CT）、磁共振血管造影 （MRA） 和 X 射线显微血管造影，要么成本高昂，需要复杂的仪器，要么在技术上具有挑战性16。2008年，Li等人描述了一种用亲脂性碳青染料DiI17标记视网膜内血管的技术。DiI掺入内皮细胞中，并通过直接扩散，染色血管膜结构，如血管生成芽和假足突17，18。由于其直接递送到内皮细胞和染料的高度荧光性质，该过程提供了强烈而持久的血管标记。2012年，Boden等人 通过 股动脉结扎后收获的大腿内收肌的全贴片成像，将DiI灌注技术应用于小鼠后肢缺血模型19。
目前的方法为评估后肢缺血和基于基因或细胞的治疗反应的新生血管形成提供了一种相对便宜且技术上可行的方法。在进一步的改编中，该协议描述了DiI灌注的应用，以高分辨率和3D在后肢坏疽的小鼠模型中对脚垫脉管系统进行成像。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Binder clips (small)</td>
			<td>Office supply store</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Buprenorphine (sustained-release)</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Butterfly needle (25 G with Luer-Lok)</td>
			<td>VWR</td>
			<td>10148-584</td>
			<td></td>
		</tr>
		<tr>
			<td>Confocal laser scanning microscope</td>
			<td>Leica</td>
			<td>TCS SP5</td>
			<td></td>
		</tr>
		<tr>
			<td>DiI (1,1&#39;-Dioctadecyl-3,3,3&#39;,3&#39;-tetramethylindocarbocyanine perchlorate)</td>
			<td>Invitrogen</td>
			<td>D282</td>
			<td></td>
		</tr>
		<tr>
			<td>Electrocautery device</td>
			<td>Gemini Cautery System</td>
			<td>5917</td>
			<td></td>
		</tr>
		<tr>
			<td>Ethanol (100%)</td>
			<td>VWR</td>
			<td>89370-084</td>
			<td></td>
		</tr>
		<tr>
			<td>Fiji (ImageJ) software</td>
			<td>NIH</td>
			<td></td>
			<td>Used version 2.1.0. Free download, no license required.</td>
		</tr>
		<tr>
			<td>Foam biopsy pads</td>
			<td>Fisher Scientific</td>
			<td>22-038-221</td>
			<td></td>
		</tr>
		<tr>
			<td>Formalin (neutral buffered, 10%)</td>
			<td>VWR</td>
			<td>89370-094</td>
			<td></td>
		</tr>
		<tr>
			<td>FVB mice</td>
			<td>Jackson Laboratory</td>
			<td>001800</td>
			<td></td>
		</tr>
		<tr>
			<td>Glucose</td>
			<td>Sigma-Aldrich</td>
			<td>G7528</td>
			<td>Used version 2.1.0.</td>
		</tr>
		<tr>
			<td>HCl (1 M)</td>
			<td>Sigma-Aldrich</td>
			<td>13-1700</td>
			<td></td>
		</tr>
		<tr>
			<td>Imaris software</td>
			<td>Oxford Instruments</td>
			<td></td>
			<td>Used version 9.6.0.</td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Pivetal</td>
			<td>NDC 46066-755-04</td>
			<td></td>
		</tr>
		<tr>
			<td>KCl</td>
			<td>Sigma-Aldrich</td>
			<td>P9333</td>
			<td></td>
		</tr>
		<tr>
			<td>Ketamine</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>L-NAME (N&#969;-Nitro-L-arginine methyl ester hydrochloride)</td>
			<td>Sigma-Aldrich</td>
			<td>N5751</td>
			<td></td>
		</tr>
		<tr>
			<td>Laser Doppler perfusion imager</td>
			<td>MoorLDI</td>
			<td>moorLDI2-HIR</td>
			<td>Used moorLDI V5 software.</td>
		</tr>
		<tr>
			<td>Microscope slides (25 x 75 x 1 mm)</td>
			<td>VWR</td>
			<td>48311-703</td>
			<td></td>
		</tr>
		<tr>
			<td>Na2HPO4</td>
			<td>Sigma-Aldrich</td>
			<td>S7907</td>
			<td></td>
		</tr>
		<tr>
			<td>NaCl</td>
			<td>Sigma-Aldrich</td>
			<td>S7653</td>
			<td></td>
		</tr>
		<tr>
			<td>NaH2PO4</td>
			<td>Sigma-Aldrich</td>
			<td>S8282</td>
			<td></td>
		</tr>
		<tr>
			<td>NaOH</td>
			<td>Sigma-Aldrich</td>
			<td>S8263</td>
			<td></td>
		</tr>
		<tr>
			<td>Needles (27 G)</td>
			<td>BD</td>
			<td>305109</td>
			<td></td>
		</tr>
		<tr>
			<td>Povidone-iodine swabstick (10%)</td>
			<td>Medline</td>
			<td>MDS093901ZZ</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical instruments</td>
			<td>Roboz Surgical</td>
			<td></td>
			<td>Fine forceps, needle driver, spring scissors, and hemostat are recommended.</td>
		</tr>
		<tr>
			<td>Suture (5-0 absorbable)</td>
			<td>DemeTECH</td>
			<td>G275017B0P</td>
			<td></td>
		</tr>
		<tr>
			<td>Syringes (10 mL)</td>
			<td>BD</td>
			<td>305482</td>
			<td></td>
		</tr>
		<tr>
			<td>Three-way stopcocks</td>
			<td>Cole-Parmer</td>
			<td>19406-49</td>
			<td></td>
		</tr>
		<tr>
			<td>Vascular Analysis Plugin</td>
			<td></td>
			<td></td>
			<td>Free download, no license required. See reference: Elfarnawany (2015).</td>
		</tr>
		<tr>
			<td>Xylazine</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

high-resolution three-dimensional imaging of the footpad vasculature in a murine hindlimb gangrene model

外周动脉疾病 （PAD） 是慢性暴露于动脉粥样硬化危险因素导致的发病的重要原因。患有最严重形式的慢性威胁肢体缺血（CLTI）的患者面临日常生活的严重损害，包括慢性疼痛，行走距离有限而没有疼痛，以及伤口未愈合。已经开发了各种动物的临床前模型来研究PAD，但小鼠后肢缺血仍然是最广泛使用的。在这些模型中，对缺血性损伤的反应可能存在显着差异，具体取决于所使用的小鼠品系以及动脉破坏的部位，数量和手段。该协议描述了一种独特的方法，将股动脉和静脉电凝与施用一氧化氮合酶（NOS）抑制剂相结合，以可靠地诱导朋友病毒B（FVB）小鼠的脚垫坏疽，类似于CLTI的组织损失。虽然仍然推荐使用传统的评估再灌注的方法，如激光多普勒灌注成像（LDPI），但亲脂性染料1，1'-二十八烷基-3，3，3'，3'-四甲基高氯吲哚花青（DiI）的心内灌注用于标记脉管系统。随后的全安装共聚焦激光扫描显微镜允许对脚垫血管网络进行高分辨率，三维（3D）重建，以补充评估后肢缺血模型中再灌注的传统方法。

该协议详细介绍了在易感FVB小鼠中使用股动脉和静脉凝固与L-NAME给药（一氧化氮合酶抑制剂）的组合诱导小鼠脚垫缺血和组织丢失的可靠方法。 图1 详细介绍了小鼠后肢脉管系统的解剖结构，并指示股动脉和静脉凝固的部位（黄色X），仅近端的外侧环反射股动脉（LCFA）和隐腘连接处的近端。需要确定LCFA，并且该结构的凝血位点在所有外科手术中保持一致。如前所述，在外?...

Watch this Scientific Journal Video about High-Resolution Three-Dimensional Imaging of the Footpad Vasculature in a Murine Hindlimb Gangrene Model at JoVE.com

High-Resolution Three-Dimensional Imaging of the Footpad Vasculature in a Murine Hindlimb Gangrene Model

本方案描述了一种独特的、临床相关的外周动脉疾病模型，该模型将股动脉和静脉电凝与施用一氧化氮合酶抑制剂相结合，以诱导FVB小鼠的后肢坏疽。然后，心内DiI灌注用于脚垫脉管系统的高分辨率三维成像。

小鼠后肢坏死模型中脚垫脉管系统的高分辨率三维成像

Peripheral arterial disease (PAD) is a significant cause of morbidity resulting from chronic exposure to atherosclerotic risk factors. Patients suffering ...

该协议介绍了如何使用质量抑制剂和股动脉凝血来创建小鼠足垫坏疽模型。然后，DiI 灌注可实现足垫脉管系统的高分辨率可视化。该模型具有临床相关性，可用于临床前药物测试和研究肢体缺血。血管系统的高分辨率可视化为评估新生血管形成提供了一种技术上方便的方法。该足垫坏疽模型在外周动脉疾病和严重肢体缺血的临床前研究中具有重要应用。DiI灌注是研究动物模型新生血管形成的有用工具。演示该程序的将是外科住院医师Antoine Ribieras和我实验室的研究助理Yulexi Ortiz。首先，在使用前立即向10毫升工作稀释剂溶液中加入200微升DiI储备溶液，制成DiI工作溶液。用手摇匀以充分混合。串联两个或三个三向旋塞阀和一个 25 号蝶针。准备10毫升注射器，其中4毫升PBS，10毫升DiI溶液和10毫升10%中性缓冲福尔马林。将带有福尔马林的注射器连接到近端流入口，并注入溶液以冲洗管路中的空气。然后转动旋塞阀以关闭端口。重复相同的过程，将注射器与DiI连接，然后PBS分别连接到中间和远端流入口。安乐死后，将动物置于吸水垫上的仰卧位，并用针头固定腋窝和下肢。使用剪刀，做一个横向切口打开腹腔，然后暴露并分割左右隔膜以进入胸腔。将胸骨两侧的胸壁从下肋骨切到第一肋骨或第二肋骨。使用止血器抓住胸骨的下端，并将其反射到动物的头部，以暴露胸腔。识别左心室，其颜色比右心室浅。用钝钳轻轻抓住心脏，将蝴蝶针插入左心室。使用剪刀或18号针刺穿右心房，让血液和灌注溶液返回心脏引流。用PBS打开注射器的端口，以每分钟一到两毫升的速度手动注射两到四毫升，持续一到两分钟，以冲洗血管系统中的血液。通过观察右心房出血确保成功灌注。注射后，关闭PBS注射器的端口。用DiI打开注射器的端口，以每分钟1至2毫升的速度注射5至10毫升，持续5分钟。注射后，关闭DiI注射器的端口并等待两分钟，以便在注射固定剂之前掺入染料。用福尔马林打开注射器的端口，以每分钟1至2毫升的速度注射5至10毫升，持续5分钟。注射后，从左心室取出针头。使用沉重的剪刀，使脚踝处的胫骨脱臼，将左右脚与小腿完全分开。将收获的脚放入装有一到两毫升10%福尔马林溶液的12孔板中。用铝箔纸包裹盘子，在四摄氏度下储存过夜。第二天，将12孔板中的固定溶液替换为每孔一到两毫升PBS。用手术刀在足底和足背部分做一个纵向切口。然后使用齿镊子和小止血器，小心地从脚和手指上取下所有皮肤，不要损坏下面的软组织。要安装组织，请将脚分别放在两个玻璃显微镜载玻片之间，两端折叠有泡沫活检垫。使用两个小活页夹将载玻片的两端压缩在一起。或者，将脚垫放回装有1至2毫升PBS的12孔板中。用铝箔覆盖后，将其储存在四摄氏度下，以保持荧光长达一个月。打开成像系统，启动采集软件，然后使用低放大倍率或低数值孔径物镜拍摄图像。单击“是”以激活阶段对话框。在配置选项卡中激活 561 纳米激光器。在主屏幕上，通过单击可见按钮激活可见光束路径。通过单击相应的活动复选框，将探测器设置为570至600纳米范围。选择采集选项卡中的磁贴扫描图标，然后设置所需的分辨率。将干燥的贴装组织样品置于显微镜载玻片之间，并将组织聚焦。导航到示例的一角。在磁贴扫描菜单下，单击标记位置按钮。导航到对面的角落，然后单击标记位置按钮，再次设置扫描边界。要设置 Z 轴的深度，请单击实时按钮。导航到样本的中心，然后使用 z 轴旋钮滚动到样本的底部。在 Z 堆栈菜单下的采集选项卡中，单击开始按钮。滚动到示例的顶部，然后单击结束按钮。单击 z 步长大小并设置为所需值。然后单击“开始”开始图像采集。该图显示了手术后一周可以从该模型中预期的组织损失变化，FABER评分记录在每张图像的右下角。重建显微镜图像显示，非结扎对照足垫的血管解剖结构正常，术后 5 天结扎后肢脚垫的灌注严重减少。手术后20天，足垫灌注已明显改善，但未达到非结扎控制的程度。该视频显示，然后使用动画功能创建了表面渲染。冲洗灌注组件中的所有气泡。如果肺在灌注过程中扩张并变成粉红色，则针头已穿透右心室，应略微缩回。肢体组织可用于免疫染色，以评估肌肉坏死、组织再生、血管密度以及祖细胞和其他组织修复细胞的募集。该小鼠坏疽模型中的DiI灌注技术已用于基因和细胞疗法的研究。该研究旨在改善严重肢体缺血患者肢体的灌注。

Research

JoVE Journal

Medicine

Human Neuroendocrine Tumor Cell Lines as a Three-Dimensional Model for the Study of Human Neuroendocrine Tumor Therapy

人类神经内分泌肿瘤细胞株为研究人类神经内分泌肿瘤治疗的三维模型

Neuroendocrine tumors (NETs) are rare tumors, with an incidence of two per 100,&#x200A;000 individuals per year, and they account for 0.5% of all human ...

科研

医学

A Murine Model of Stent Implantation in the Carotid Artery for the Study of Restenosis

颈动脉支架植入再狭窄的研究的小鼠模型

Despite  the considerable progress made in the stent development in the last decades,  cardiovascular diseases remain the main cause of death in western ...

Three-dimensional Imaging of Nociceptive Intraepidermal Nerve Fibers in Human Skin Biopsies

三维成像伤害表皮内神经纤维在人体皮肤活检

A punch biopsy of the skin is commonly used to quantify intraepidermal nerve fiber densities (IENFD) for the diagnosis of peripheral polyneuropathy 1,2. ...

Osmotic Drug Delivery to Ischemic Hindlimbs and Perfusion of Vasculature with Microfil for Micro-Computed Tomography Imaging

药物输送到微型计算机断层扫描成像缺血后肢血管灌注与微滤渗透

Preclinical research in animal models of peripheral arterial disease plays a vital role in testing the efficacy of therapeutic agents designed to ...

Quantification of Breast Cancer Cell Invasiveness Using a Three-dimensional (3D) Model

Quantification of Breast Cancer Cell Invasiveness Using a Three-dimensional 3D Model

乳腺癌细胞侵袭定量使用三维（3D）模型

It is now well known that the cellular and tissue microenvironment are critical regulators influencing tumor initiation and progression. Moreover, the ...

Three-dimensional Co-culture Model for Tumor-stromal Interaction

对于肿瘤基质相互作用的三维共培养模式

Cancer progression (initiation, growth, invasion and metastasis) occurs through interactions between malignant cells and the surrounding tumor stromal ...

Three-Dimensional (3D) Tumor Spheroid Invasion Assay

Three-Dimensional 3D Tumor Spheroid Invasion Assay

三维（3D）肿瘤球体侵袭试验

Invasion of surrounding normal tissues is generally considered to be a key hallmark of malignant (as opposed to benign) tumors. For some cancers in ...

Quantitative Visualization of Leukocyte Infiltrate in a Murine Model of Fulminant Myocarditis by Light Sheet Microscopy

通过轻型显微镜对小鼠急性心肌炎模型中白细胞浸润的定量可视化

Light-sheet fluorescence microscopy (LSFM), in combination with chemical clearing protocols, has become the gold standard for analyzing fluorescently ...

Electrophysiological Assessment of Murine Atria with High-Resolution Optical Mapping

高分辨率光学制图对小鼠心房电生理的评价

Recent genome-wide association studies targeting atrial fibrillation (AF) have indicated a strong association between the genotype and ...

Assessing Therapeutic Angiogenesis in a Murine Model of Hindlimb Ischemia

评估下肢缺血的Murine模型的治疗血管生成

Critical limb ischemia (CLI) is a serious condition that entails a high risk of lower limb amputation. Despite revascularization being the gold-standard ...