Name: precision cut lung slices as an efficient tool for ex vivo pulmonary vessel structure and contractility studies
Uploaded: 2021-05-24T13:00:00
Description: Watch this Scientific Journal Video about Precision Cut Lung Slices as an Efficient Tool for Ex vivo Pulmonary Vessel Structure and Contractility Studies at JoVE.com

作者要感谢袁昊博士和刘开峰博士的技术支持。这项工作得到了NIH 1R01 HL150106-01A1，Parker B. Francis奖学金和肺动脉高压协会Aldrighetti研究奖的支持。

所有动物护理均符合波士顿儿童医院的指导方针和机构动物护理和使用委员会批准的协议。本研究中使用的小鼠是野生型C57 / B6小鼠和Cdh5-CreERT2 x Ai14 tdTomato杂交小鼠。
1. 溶液的制备
<ol><li>提前准备实验期间所需的磷酸盐缓冲溶液（1x PBS）和2%琼脂糖溶液。<ol>
<li>将 2 克琼脂糖粉混合到 100 mL 高压灭菌水中。在微波炉中加热几秒钟，直到溶液变清。</li>
		<li>将溶液置于42°C的水...

<ol>
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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=Preparation+and+incubation+of+precision-cut+liver+and+intestinal+slices+for+application+in+drug+metabolism+and+toxicity+studies.">Preparation and incubation of precision-cut liver and intestinal slices for application in drug metabolism and toxicity studies.</a> Nature Protocols. 5 (9), 1540-1551 (2010).</li><li>Alsafadi, H. N., 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=Applications+and+approaches+for+three-dimensional+precision-cut+lung+slices.+Disease+modeling+and+drug+discovery.">Applications and approaches for three-dimensional precision-cut lung slices. Disease modeling and drug discovery.</a> American Journal of Respiratory Cell and Molecular Biology. 62 (6), 681-691 (2020).</li><li>Morin, J. 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=Precision+cut+lung+slices+as+an+efficient+tool+for+in+vitro+lung+physio-pharmacotoxicology+studies.">Precision cut lung slices as an efficient tool for in vitro lung physio-pharmacotoxicology studies.</a> Xenobiotica. 43 (1), 63-72 (2013).</li><li>Springer, J., Fischer, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Substance+P-induced+pulmonary+vascular+remodelling+in+precision+cut+lung+slices.">Substance P-induced pulmonary vascular remodelling in precision cut lung slices.</a> The European Respiratory Journal. 22 (4), 596-601 (2003).</li><li>Suleiman, 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=Argon+reduces+the+pulmonary+vascular+tone+in+rats+and+humans+by+GABA-receptor+activation.">Argon reduces the pulmonary vascular tone in rats and humans by GABA-receptor activation.</a> Scientific Reports. 9 (1), 1902 (2019).</li><li>Rieg, A. 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=Cardiovascular+agents+affect+the+tone+of+pulmonary+arteries+and+veins+in+precision-cut+lung+slices.">Cardiovascular agents affect the tone of pulmonary arteries and veins in precision-cut lung slices.</a> PLoS One. 6 (12), 29698 (2011).</li><li>Perez, J. F., Sanderson, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+frequency+of+calcium+oscillations+induced+by+5-HT,+ACH,+and+KCl+determine+the+contraction+of+smooth+muscle+cells+of+intrapulmonary+bronchioles.">The frequency of calcium oscillations induced by 5-HT, ACH, and KCl determine the contraction of smooth muscle cells of intrapulmonary bronchioles.</a> The Journal of General Physiology. 125 (6), 535-553 (2005).</li><li>Deng, C. 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=Upregulation+of+5-hydroxytryptamine+receptor+signaling+in+coronary+arteries+after+organ+culture.">Upregulation of 5-hydroxytryptamine receptor signaling in coronary arteries after organ culture.</a> PLoS One. 9 (9), 107128 (2014).</li><li>Sandker, S. 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=Adventitial+dissection:+A+simple+and+effective+way+to+reduce+radial+artery+spasm+in+coronary+bypass+surgery.">Adventitial dissection: A simple and effective way to reduce radial artery spasm in coronary bypass surgery.</a> Interactive Cardiovascular and Thoracic Surgery. 17 (5), 784-789 (2013).</li><li>Naik, J. 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=Pressure-induced+smooth+muscle+cell+depolarization+in+pulmonary+arteries+from+control+and+chronically+hypoxic+rats+does+not+cause+myogenic+vasoconstriction.">Pressure-induced smooth muscle cell depolarization in pulmonary arteries from control and chronically hypoxic rats does not cause myogenic vasoconstriction.</a> Journal of Applied Physiology. 98 (3), 1119-1124 (2005).</li><li>Lopez-Lopez, J. G., 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=Diabetes+induces+pulmonary+artery+endothelial+dysfunction+by+NADPH+oxidase+induction.">Diabetes induces pulmonary artery endothelial dysfunction by NADPH oxidase induction.</a> American Journal of Physiology. Lung Cellular and Molecular Physiology. 295 (5), 727-732 (2008).</li><li>Gonzalez-Tajuelo, 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=Spontaneous+pulmonary+hypertension+associated+with+systemic+sclerosis+in+P-selectin+glycoprotein+Ligand+1-deficient+mice.">Spontaneous pulmonary hypertension associated with systemic sclerosis in P-selectin glycoprotein Ligand 1-deficient mice.</a> Arthritis &amp; Rheumatology. 72 (3), 477-487 (2020).</li><li>Bai, Y., Sanderson, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modulation+of+the+Ca2++sensitivity+of+airway+smooth+muscle+cells+in+murine+lung+slices.">Modulation of the Ca2+ sensitivity of airway smooth muscle cells in murine lung slices.</a> American Journal of Physiology. Lung Cellular and Molecular Physiology. 291 (2), 208-221 (2006).</li><li>Nishiyama, S. 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=Vascular+function+and+endothelin-1:+tipping+the+balance+between+vasodilation+and+vasoconstriction.">Vascular function and endothelin-1: tipping the balance between vasodilation and vasoconstriction.</a> Journal of Applied Physiology. 122 (2), 354-360 (2017).</li><li>Schneider, M. P., Inscho, E. W., Pollock, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Attenuated+vasoconstrictor+responses+to+endothelin+in+afferent+arterioles+during+a+high-salt+diet.">Attenuated vasoconstrictor responses to endothelin in afferent arterioles during a high-salt diet.</a> American Journal of Physiology. Renal Physiology. 292 (4), 1208-1214 (2007).</li><li>Inscho, E. W., Imig, J. D., Cook, A. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Afferent+and+efferent+arteriolar+vasoconstriction+to+angiotensin+II+and+norepinephrine+involves+release+of+Ca2++from+intracellular+stores.">Afferent and efferent arteriolar vasoconstriction to angiotensin II and norepinephrine involves release of Ca2+ from intracellular stores.</a> Hypertension. 29, 222-227 (1997).</li><li>Vecchione, 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=Protection+from+angiotensin+II-mediated+vasculotoxic+and+hypertensive+response+in+mice+lacking+PI3Kgamma.">Protection from angiotensin II-mediated vasculotoxic and hypertensive response in mice lacking PI3Kgamma.</a> The Journal of Experimental Medicine. 201 (8), 1217-1228 (2005).</li></ol>

在这份手稿中，描述了一种增强的方法，以产生高分辨率的小鼠肺组织图像，该图像保留了血管结构并优化了实验灵活性，特别是使用PCLS的应用来获得可以在三维空间中观察的肺组织微切片，并保留脉管系统的收缩力。使用活性试剂，该方案表明，精心准备和保存的切片可以保持活力超过一周。微切片的存活力得以在脉管系统和气道结构上进行多次和长时间的实验，使其成为测试多种药物 的?...

在不牺牲解剖结构的情况下保留细胞成分的肺组织的制备和成像的进步提供了对肺部疾病的详细了解。在保持生理结构的同时识别蛋白质，RNA和其他生物化合物的能力提供了有关细胞空间排列的重要信息，可以拓宽对许多肺部疾病的病理生理学的理解。当应用于动物模型时，这些详细的图像可以导致对肺血管疾病（如肺动脉高血压）的更好理解，从而可能导致治疗策略的改进。
尽管技术进步，但获得小鼠肺组织的高质量图像仍然是一个挑战。呼吸周期由吸入期间产生的负胸内压驱动1。传统上，当获得活检并准备肺部样本进行成像时，负压梯度丢失，导致气道和脉管系统塌陷，不再代表自身的当前状态。为了获得反映当前状况的真实图像，必须重新充气肺气道，并灌注脉管系统，将动态肺变成静态夹具。这些不同技术的应用可以保持结构完整性，肺脉管系统和细胞成分，包括免疫细胞，如巨噬细胞，允许将肺组织视为尽可能接近其生理状态。
精密切肺切片（PCLS）是研究肺血管系统解剖学和生理学的理想工具2。PCLS提供三维肺组织的详细成像，同时保留结构和细胞成分。PCLS已被用于动物和人类模型中，以允许在三维空间中提供细胞功能的实时高分辨率图像，使其成为研究潜在治疗靶点，测量小气道收缩和研究慢性肺部疾病（如COPD，ILD和肺癌）的病理生理学的理想工具3。使用类似的技术，PCLS样品暴露于血管收缩剂可以保持肺结构和血管收缩性，复制 体外 条件。除了保持收缩性外，制备的样品在正确制备时还可以进行额外的分析，例如RNA测序，蛋白质印迹和流式细胞术。最后，在肺部收获后用tdTomato荧光标记的报告色标记细胞可以在制备微切片后保留标记，使其成为细胞追踪研究的理想选择。这些技术的整合提供了一个复杂的模型，保留了细胞的空间排列和血管收缩力，可以更详细地了解肺脉管系统疾病的信号级联反应和潜在的治疗方案。
在这份手稿中，PCLS小鼠肺组织暴露于血管收缩剂，显示出保留的结构完整性和血管收缩性。该研究表明，适当准备和处理的组织可以保持活力10天。该研究还证明了用内源性荧光（tdTomato）保存细胞，允许样品提供肺脉管系统和结构的高分辨率图像。最后，已经描述了处理和制备用于RNA测量的组织切片和蛋白质印迹以研究潜在机制的方法。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>0.5cc of fractionated heparin in syringe</td>
			<td>BD</td>
			<td>100 USP units per mL</td>
			<td></td>
		</tr>
		<tr>
			<td>1X PBS</td>
			<td>Corning</td>
			<td>&#160;21-040-CM</td>
			<td></td>
		</tr>
		<tr>
			<td>20 1/2 inch gauge blunt end needle for trachea cannulation</td>
			<td>Cml Supply</td>
			<td>90120050D</td>
			<td></td>
		</tr>
		<tr>
			<td>30cc syringe</td>
			<td>BD</td>
			<td>309650</td>
			<td></td>
		</tr>
		<tr>
			<td>Anti Anti solution</td>
			<td>Gibco</td>
			<td>15240096</td>
			<td></td>
		</tr>
		<tr>
			<td>Automated vibrating blade microtome</td>
			<td>Leica</td>
			<td>VT1200S</td>
			<td></td>
		</tr>
		<tr>
			<td>Cell Viability Reagent (alamarBlue)</td>
			<td>Thermofisher</td>
			<td>DAL1025</td>
			<td></td>
		</tr>
		<tr>
			<td>Confocal</td>
			<td>Zeiss</td>
			<td>880</td>
			<td></td>
		</tr>
		<tr>
			<td>Dulbecco&#8217;s Modified Eagle Medium and GLutaMAX, supplemented with 10% FBS, 1% Pen/Strep</td>
			<td>Gibco</td>
			<td>10569-010</td>
			<td></td>
		</tr>
		<tr>
			<td>Endothelin-1</td>
			<td>Sigma</td>
			<td>E7764</td>
			<td></td>
		</tr>
		<tr>
			<td>KCl</td>
			<td>Sigma</td>
			<td>7447-40-7</td>
			<td></td>
		</tr>
		<tr>
			<td>Mortar and Pestle</td>
			<td>Amazon</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>RIPA lysis and extraction buffer</td>
			<td>Thermoscientific</td>
			<td>89900</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical suture 6/0</td>
			<td>FST</td>
			<td>18020-60</td>
			<td></td>
		</tr>
		<tr>
			<td>TRIzol Reagent</td>
			<td>Invitrogen, Thermofisher</td>
			<td>15596026</td>
			<td></td>
		</tr>
		<tr>
			<td>UltraPure Low Melting Point Agarose</td>
			<td>Invitrogen</td>
			<td>16520050</td>
			<td></td>
		</tr>
		<tr>
			<td>Vibratome</td>
			<td>Leica Biosystems</td>
			<td>VT1200 S</td>
			<td></td>
		</tr>
		<tr>
			<td>Winged blood collection set (Butterfly needle) 25-30G</td>
			<td>BD</td>
			<td>25-30G</td>
			<td></td>
		</tr>
	</tbody>
</table>

precision cut lung slices as an efficient tool for ex vivo pulmonary vessel structure and contractility studies

鼠肺组织的可视化提供了有关潜在气道和脉管系统的有价值的结构和细胞信息。然而，真正代表生理状况的肺血管的保存仍然存在挑战。此外，鼠肺的精细配置导致技术挑战，为保留细胞组成和结构的高质量图像制备样品。类似地，可以进行细胞收缩力测定来研究细胞 在体外对血管收缩剂作出反应的潜力，但这些测定不能再现完整肺的复杂环境。与这些技术问题相反，精确切割肺切片（PCLS）方法可以作为一种有效的替代方案，在没有区域偏倚的情况下以三维方式可视化肺组织，并作为长达10天的实时替代收缩力模型。使用PCLS制备的组织保留了结构和空间取向，使其成为 离体研究疾病过程的理想选择。从诱导的tdTomato报告小鼠模型中收获的PCLS中内源性tdTomato标记细胞的位置可以通过共聚焦显微镜成功可视化。暴露于血管收缩剂后，PCLS显示出血管收缩性和肺结构的保存，这可以通过延时模块捕获。结合其他程序，如蛋白质印迹和RNA分析，PCLS有助于全面了解各种疾病背后的信号级联反应，并导致更好地理解肺血管疾病的病理生理学。

当添加到细胞或组织中时，活力试剂被活组织的还原环境修饰并变成粉红色/红色，变得高度荧光。 图3显示了从第0-1天和第9-10天检测到的代表性颜色变化。如前所述，溶液在一夜之间开始变为蓝色并变成粉红色，显示出可行性。颜色变化通常发生在1-4小时内;但是，可能需要更长的时间。为了测定活力，使用读板器测定振动切片机制备的样品和4%PFA固定肺切片的吸光度，作为...

Watch this Scientific Journal Video about Precision Cut Lung Slices as an Efficient Tool for Ex vivo Pulmonary Vessel Structure and Contractility Studies at JoVE.com

Precision Cut Lung Slices as an Efficient Tool for Ex vivo Pulmonary Vessel Structure and Contractility Studies

这里介绍的是用于保存PCLS小鼠肺组织的血管收缩力的方案，从而产生肺脉管系统和气道的复杂三维图像，可以保存长达10天，容易受到许多程序的影响。

精密切肺切片作为 离体 肺血管结构和收缩力研究的有效工具

The visualization of murine lung tissue provides valuable structural and cellular information regarding the underlying airway and vasculature. However, ...

精密切肺切片法将保持三维结构和空间方向，无区域偏差，可作为血管和气道收缩力测量的活体替代模型。结合延时成像、蛋白质印迹和 RNA 分析，精确切开的肺切片有助于全面了解信号级联反应，这些级联反应是各种疾病的基础，并有助于更好地了解肺血管疾病的病理生理学。首先，将安乐死的成年小鼠置于仰卧位置，并用胶带将爪子和鼻子贴在桌子上，将其固定到位。用70%乙醇喷洒小鼠的腹侧表面。使用镊子抬起小鼠腹部皮肤在膀胱的位置，并使用手术剪刀在中线处切开，向上移动到颈部区域以暴露气管。然后用镊子抬起下面的筋膜层，用手术剪刀暴露内脏器官和纵隔室，再次从下切到上。切开中线的胸骨，露出心脏和肺部。从肝脏和腹部隔室仔细解剖隔膜。找到肠下腔静脉并切割。定位右心室后，使用25至30号蝶针将0.5毫升1%分馏肝素注射到右心室中，并用10至15毫米PBS缓慢但稳定地冲洗。定位喉部后，使用钝头镊子解剖周围的筋膜和组织，然后将手术缝合线放在气管下方并松散地打结。在气管上放一个高于琴弦的小孔，并用20号钝头针插管。使用手术结收紧针头和气管周围的缝合线。然后将2.5至4毫升琼脂糖注射到气管中并监测肺部的充气。注射琼脂糖后，将冷PBS倒在肺部以固化琼脂糖。然后切除手术缝合线上的气管，并通过去除任何粘连和筋膜将肺和心脏从纵隔上解剖。凝固后，将DMEM中的组织放在冰上的培养皿中，然后立即使用振动机切开一个叶片。为了准备肺切片，用超级胶水将样品连接到平台上，保持内侧朝上。然后用 PBS 填充样品容器。在容器中装满冰块，以保持周围的PBS和样品冷却。打开振动器。将厚度调整为300微米，频率调整为100赫兹，振幅调整为0.6毫米，速度调整为每秒5微米。使用内六角扳手，将刀片架转到安全位置，然后打开钳口插入刀片。然后用内六角扳手拧紧钳口。将样品放在盒子上，确保其浸没在PBS中。将盒子向上滑动到正确的位置，并确保其牢固。将刀片架转入适当的位置进行切片。将振动片向上移动到块的边缘，然后手动降低刀片，直到它与样品顶部均匀。确认设置并运行振动器。切开新鲜切片后，从PBS中取出样品，并将其放入含有10毫升含有一次性抗真菌抗生素的DMEM的无菌培养皿中。将所有切片放入培养皿中后，将刀片完全缩回，并使用内六角扳手将刀片抬高到安全位置。将样品储存在37摄氏度的DMEM中。并将振动切片片放在显微镜载玻片上。使用清洁湿巾去除多余的介质。将样品放在相差显微镜上，并使用10至20倍放大倍率来识别容器。然后加入500微升血管收缩剂，例如60毫摩尔氯化钾或一微摩尔内皮素-1以完全浸没载玻片。通过录制30到60秒来观察和捕获视频。在开始实验之前，用一升液氮填充一个小的聚苯乙烯泡沫盒，并在装有液氮的容器中放置两个1.5毫升的微量离心管。将四到五片新鲜的振动切片放入砂浆碗中。将5至10毫升液氮倒在切片上，并在液氮蒸发之前快速使用杵将切片研磨成粉末。使用预冷的钢制实验室铲斗将粉末舀入两个冷冻的微量离心管中，并通过加入500微升用于RNA分离的RNA提取试剂或500微升用于蛋白质裂解的RIPA缓冲液来裂解粉末。在代表性的可行性实验中，从第0天到第1天和第9天到第10天检测到PCLS切片的颜色变化。该溶液最初是蓝色的，一夜之间变成粉红色，显示出可行性。颜色变化通常发生在一到四个小时内，但可能需要更长的时间。此处显示了振动切片机制备的组织在562和613纳米波长处的吸光度的每日差异。使用内皮素-1和氯化钾的振动切片机制备的肺组织中脉管系统的收缩可以在这里可视化。在他莫昔芬注射后一周的细胞中保存tdTomato标记在这些肺切片中得到了证明。最重要的是，在进行额外的实验和成像之前，要很好地进行肺部清除和充气步骤，以从循环中去除所有血液。除血管疾病外，PCLS还可用于研究气道相关疾病的生理生物学，并可在临床相关情况下在人体肺组织上进一步测试。

Research

JoVE Journal

Medicine

Precision Cut Lung Slices as an Efficient Tool for Ex vivo Pulmonary Vessel Structure and Contractility Studies

Videomorphometric Analysis of Hypoxic Pulmonary Vasoconstriction of Intra-pulmonary Arteries Using Murine Precision Cut Lung Slices

使用穆林精密切肺片对肺动脉内下氧肺血管收缩进行视频形态分析

Acute alveolar hypoxia causes pulmonary vasoconstriction (HPV) - also known as von Euler-Liljestrand mechanism - which serves to match lung perfusion to ...

科研

医学

Fundus Photography as a Convenient Tool to Study Microvascular Responses to Cardiovascular Disease Risk Factors in Epidemiological Studies

眼底摄影作为一种方便的工具，研究微血管回应流行病学研究心血管疾病的风险因素

The microcirculation consists of blood vessels with diameters less than 150 &#181;m. It makes up a large part of the circulatory system and plays an ...

Method of Isolated Ex Vivo Lung Perfusion in a Rat Model: Lessons Learned from Developing a Rat EVLP Program

Method of Isolated Ex Vivo Lung Perfusion in a Rat Model: Lessons Learned from Developing a Rat EVLP Program

的隔离方法<em&gt;离体</em&gt;肺灌注的大鼠模型：教训制定鼠EVLP课程中学

The number of acceptable donor lungs available for lung transplantation is severely limited due to poor quality. Ex-Vivo Lung Perfusion (EVLP) has allowed ...

Ex vivo Live Imaging of Lung Metastasis and Their Microenvironment

Ex vivo Live Imaging of Lung Metastasis and Their Microenvironment

体外肺转移及微环境的实时成像

Metastasis is a major cause for cancer-related morbidity and mortality. Metastasis is a multistep process and due to its complexity, the exact cellular ...

The 4-vessel Sampling Approach to Integrative Studies of Human Placental Physiology In Vivo

The 4-vessel Sampling Approach to Integrative Studies of Human Placental Physiology In Vivo

人体胎盘生理学综合研究的4血管取样方法<em&gt;体内</em

The human placenta is highly inaccessible for research while still in utero. The current understanding of human placental physiology in vivo is therefore ...

Combining Volumetric Capnography And Barometric Plethysmography To Measure The Lung Structure-function Relationship

体积摄影与气压面学相结合测量肺结构-功能关系

Tools to measure lung and airways volume are critical for pulmonary researchers interested in evaluating the impact of disease or novel therapies on the ...

Ex Vivo Corneal Organ Culture Model for Wound Healing Studies

创面愈合研究的前活体角膜器官培养模型

The cornea has been used extensively as a model system to study wound healing. The ability to generate and utilize primary mammalian cells in two ...

Quantitative Mapping of Specific Ventilation in the Human Lung using Proton Magnetic Resonance Imaging and Oxygen as a Contrast Agent

以质子磁共振成像和氧为造影剂对人肺特异性通气进行定量定位

Specific ventilation imaging (SVI) is a functional magnetic resonance imaging technique capable of quantifying specific ventilation &#8213; the ratio of ...

Optocardiography and Electrophysiology Studies of Ex Vivo Langendorff-perfused Hearts

前维沃·兰根多夫注入心脏的心电生理学研究

Small animal models are most commonly used in cardiovascular research due to the availability of genetically modified species and lower cost compared to ...

Murine Precision-Cut Liver Slices as an Ex Vivo Model of Liver Biology

毛利精密切肝片作为肝脏生物学的Ex Vivo模型

Understanding the mechanisms of liver injury, hepatic fibrosis, and cirrhosis that underlie chronic liver diseases (i.e., viral hepatitis, non-alcoholic ...

精密切肺切片作为 离体 肺血管结构和收缩力研究的有效工具

精密切肺切片作为离体肺血管结构和收缩力研究的有效工具