Name: murine excisional wound healing model and histological morphometric wound analysis
Uploaded: 2020-08-21T15:00:00
Description: Watch this Scientific Journal Video about Murine Excisional Wound Healing Model and Histological Morphometric Wound Analysis at JoVE.com

我们感谢邓恩瓦尔德实验室的所有成员，他们多年来为优化该协议做出了贡献，并感谢吉娜·沙特曼，他坚持推广使用串行分段进行伤口分析，使该协议的创建成为可能。这项工作得到了国家卫生和、国家艾滋病和国家卫生系统向马丁·邓恩瓦尔德（AR067739）的资助。

所有实验都按照联邦法规完成，爱荷华大学的政策和程序已获爱荷华大学 IACUC 批准。
1. 畜牧业
<ol><li>在毛囊阶段为细胞原时，在8-10周大时使用所需小鼠线的成年小鼠。</li>
	<li>在手术当天，将老鼠分开进入干净的笼子和单独的房子，以尽量减少伤口的破坏。</li>
</ol>2. 手术
注：没有必要保持无菌的手术条件。虽然应注意保持动物之间的...

<ol>
	<li>Eming, S. A., Martin, P., Tomic-Canic, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Wound+repair+and+regeneration:+mechanisms,+signaling,+and+translation.">Wound repair and regeneration: mechanisms, signaling, and translation.</a> Science Translational Medicine. 6 (265), (2014).</li><li>Park, 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=Tissue-scale+coordination+of+cellular+behaviour+promotes+epidermal+wound+repair+in+live+mice.">Tissue-scale coordination of cellular behaviour promotes epidermal wound repair in live mice.</a> Nature Cell Biology. 19 (2), 155-163 (2017).</li><li>Gurtner, G. C., Werner, S., Barrandon, Y., Longaker, M. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Wound+repair+and+regeneration.">Wound repair and regeneration.</a> Nature. 453 (7193), 314-321 (2008).</li><li>Ansell, D. M., Campbell, L., Thomason, H. A., Brass, A., Hardman, 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=A+statistical+analysis+of+murine+incisional+and+excisional+acute+wound+models.">A statistical analysis of murine incisional and excisional acute wound models.</a> Wound Repair Regeneration. 22 (2), 281-287 (2014).</li><li>Elliot, S., Wikramanayake, T. C., Jozic, I., Tomic-Canic, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Modeling+Conundrum:+Murine+Models+for+Cutaneous+Wound+Healing.">A Modeling Conundrum: Murine Models for Cutaneous Wound Healing.</a> Journal of Investigative Dermatology. 138 (4), 736-740 (2018).</li><li>Crowe, M. J., Doetschman, T., Greenhalgh, D. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Delayed+wound+healing+in+immunodeficient+TGF-beta+1+knockout+mice.">Delayed wound healing in immunodeficient TGF-beta 1 knockout mice.</a> Journal of Investigative Dermatology. 115 (1), 3-11 (2000).</li><li>Pietramaggiori, 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=Improved+cutaneous+healing+in+diabetic+mice+exposed+to+healthy+peripheral+circulation.">Improved cutaneous healing in diabetic mice exposed to healthy peripheral circulation.</a> Journal of Investigative Dermatology. 129 (9), 2265-2274 (2009).</li><li>Cold Spring Harbor. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Paraformaldehyde+in+PBS.">Paraformaldehyde in PBS.</a> Cold Spring Harbor Protocols. 1 (1), (2006).</li><li>Gerharz, 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=Morphometric+analysis+of+murine+skin+wound+healing:+standardization+of+experimental+procedures+and+impact+of+an+advanced+multitissue+array+technique.">Morphometric analysis of murine skin wound healing: standardization of experimental procedures and impact of an advanced multitissue array technique.</a> Wound Repair Regeneration. 15 (1), 105-112 (2007).</li><li>Colwell, A. S., Krummel, T. M., Kong, W., Longaker, M. T., Lorenz, H. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Skin+wounds+in+the+MRL/MPJ+mouse+heal+with+scar.">Skin wounds in the MRL/MPJ mouse heal with scar.</a> Wound Repair Regeneration. 14 (1), 81-90 (2006).</li><li>Le, 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=Transforming+growth+factor+beta+3+is+required+for+proper+excisional+wound+repair+in+vivo.">Transforming growth factor beta 3 is required for proper excisional wound repair in vivo.</a> PLoS One. 7 (10), 48040 (2012).</li><li>Sato, T., Yamamoto, M., Shimosato, T., Klinman, 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=Accelerated+wound+healing+mediated+by+activation+of+Toll-like+receptor+9.">Accelerated wound healing mediated by activation of Toll-like receptor 9.</a> Wound Repair Regeneration. 18 (6), 586-593 (2010).</li><li>Martin, P., Leibovich, S. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inflammatory+cells+during+wound+repair:+the+good,+the+bad+and+the+ugly.">Inflammatory cells during wound repair: the good, the bad and the ugly.</a> Trends in Cell Biology. 15 (11), 599-607 (2005).</li><li>Shaw, T. J., Martin, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Wound+repair:+a+showcase+for+cell+plasticity+and+migration.">Wound repair: a showcase for cell plasticity and migration.</a> Current Opinion in Cell Biology. 42, 29-37 (2016).</li><li>Hoffman, M., Monroe, 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=Low+intensity+laser+therapy+speeds+wound+healing+in+hemophilia+by+enhancing+platelet+procoagulant+activity.">Low intensity laser therapy speeds wound healing in hemophilia by enhancing platelet procoagulant activity.</a> Wound Repair Regeneration. 20 (5), 770-777 (2012).</li><li>Tomasek, J. J., Gabbiani, G., Hinz, B., Chaponnier, C., Brown, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Myofibroblasts+and+mechano-regulation+of+connective+tissue+remodelling.">Myofibroblasts and mechano-regulation of connective tissue remodelling.</a> Nature Reviews Molecular Cell Biology. 3 (5), 349-363 (2002).</li><li>Uchiyama, 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=SOX2+Epidermal+Overexpression+Promotes+Cutaneous+Wound+Healing+via+Activation+of+EGFR/MEK/ERK+Signaling+Mediated+by+EGFR+Ligands.">SOX2 Epidermal Overexpression Promotes Cutaneous Wound Healing via Activation of EGFR/MEK/ERK Signaling Mediated by EGFR Ligands.</a> Journal of Investigative Dermatology. 139 (8), 1809-1820 (2019).</li><li>Sen, C. 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=Human+skin+wounds:+a+major+and+snowballing+threat+to+public+health+and+the+economy.">Human skin wounds: a major and snowballing threat to public health and the economy.</a> Wound Repair Regeneration. 17 (6), 763-771 (2009).</li><li>Rhea, L., 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=Interferon+regulatory+factor+6+is+required+for+proper+wound+healing+in+vivo.">Interferon regulatory factor 6 is required for proper wound healing in vivo.</a> Developmental Dynamics. 249 (4), 509-522 (2020).</li></ol>

双边切除伤口模型是一种高度可定制的程序，可用于研究伤口愈合的许多不同的方面。在开始伤口愈合项目之前，调查人员应执行功率分析，以确定检测特定效果大小的缺陷所需的伤口数量。文献中对于单个小鼠或伤口是否应用作生物复制，存在不一致之处，然而，最近的一项研究表明，单个动物4上的两个伤口之间没有显著的相关性。这表明，单个动物的伤口彼此独立，可以被...

皮肤伤口愈合是一个复杂的生物过程，连续重叠阶段。它要求协调在时间和空间上调节的细胞和分子过程，以恢复受损上皮的屏障功能。在第一阶段，炎症，嗜中性粒细胞和巨噬细胞迁移到伤口，调动局部和全身防御1。炎症阶段的跟随和重叠是增殖阶段。成纤维细胞开始迅速增殖并迁移到造粒组织中。角膜细胞远离前缘方向扩散到伤口，因为前缘的分化角细胞迁移至伤口2的再上皮化。最后，改造和成熟阶段开始，在此期间，造粒组织中的成纤维细胞开始合成和沉积胶原蛋白。新矩阵的改造和组织可以持续长达1年后受伤3。由于涉及多种细胞类型交谈的重叠事件的复杂性，尽管经过多年的研究，许多细胞和分子机制的基础伤口愈合仍然不太了解。
小鼠模型是研究伤口愈合机制的主要哺乳动物模型，由于其易用性，相对较低的成本和遗传操纵性1，4，5。虽然在穆林模型中描述了不同类型的伤口，但最常见的是切除伤口（双边冲孔或直接打孔活检），其次是切口伤口模型4。切除伤口模型比切口模型有明显的优势，因为它固有的产生控制组织，没有经过愈合过程。作为手术方案的一部分切除的冲孔活检组织可以与受伤组织相同的方式进行处理，并用于确定所需标准的恒时条件。如果评估皮肤预处理的效果或在受伤时确认成功的基因改变，可切除控制组织也很有用。
治疗参数可以通过许多不同的技术进行评估，包括平面学或组织学。然而，平面测量只能评估伤口的可见特征，由于有疤痕的存在，往往与组织学所可视化的愈合测量不相关，从而使组织学成为分析4的"黄金标准"。尽管组织学分析是金本位标准，但它通常对伤口6、7的任意子集进行。例如，在嵌入和切段伤口之前将伤口切割到"一半"是目前常见的做法，以减少用于分段材料和数据分析所花费的时间和资源。本协议中描述的形态分析方法被开发为包括整个伤口组织，准确反映伤口的形态特征，并增加检测伤口愈合缺陷的可能性，效果较小。在该协议中，我们详细介绍了产生最常研究的穆林伤口的手术方法、双边全厚切除伤口，以及一种详细而严格的组织学分析方法，这种方法在实地很少使用。

<table>
	<tbody>
		<tr>
			<td>100% ethanol</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>70% ethanol</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>80% ethanol</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>95% ethanol</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Alcohol Prep</td>
			<td>NOVAPLUS</td>
			<td>V9100</td>
			<td>70% Isopropyl alcohol, sterile</td>
		</tr>
		<tr>
			<td>Ammonium hydroxide</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Biopsy pads</td>
			<td>Cellpath</td>
			<td>22-222-012</td>
			<td></td>
		</tr>
		<tr>
			<td>Black plastic sheet</td>
			<td></td>
			<td></td>
			<td>Something firm yet manipulatable about the size of a sheet of paper</td>
		</tr>
		<tr>
			<td>Brightfield microscope</td>
			<td></td>
			<td></td>
			<td>With digital acquisition capabilities and a 4X objective</td>
		</tr>
		<tr>
			<td>Cotton tipped applicators</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Coverslips</td>
			<td></td>
			<td></td>
			<td>22 x 60 #1</td>
		</tr>
		<tr>
			<td>Dental wax sheets</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Digital camera</td>
			<td></td>
			<td></td>
			<td>Include a ruler for scale, if applicable</td>
		</tr>
		<tr>
			<td>Dissection teasing needle (straight)</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Embedding molds</td>
			<td></td>
			<td></td>
			<td>22 x 22 x 12</td>
		</tr>
		<tr>
			<td>Embedding rings</td>
			<td>Simport Scientific Inc.</td>
			<td>M460</td>
			<td></td>
		</tr>
		<tr>
			<td>Eosin Y</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Glacial acetic acid</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Hair clipper</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Heating pad</td>
			<td>Conair</td>
			<td></td>
			<td>Moist dry Heating Pad</td>
		</tr>
		<tr>
			<td>Hematoxylin</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Microtome</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Microtome blades</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Paint brushes</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
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			<td>Paraffin Type 6</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Paraformaldehyde</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Permount</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Phosphate buffer solution (PBS)</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Povidone-iodine</td>
			<td>Aplicare</td>
			<td>82-255</td>
			<td></td>
		</tr>
		<tr>
			<td>Processing cassette</td>
			<td>Simport Scientific Inc.</td>
			<td>M490-2</td>
			<td></td>
		</tr>
		<tr>
			<td>Razor blades</td>
			<td>ASR</td>
			<td></td>
			<td>.009 Regular Duty</td>
		</tr>
		<tr>
			<td>Scalpel blades #10</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Scalpel handle</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Sharp surgical scissors</td>
			<td></td>
			<td></td>
			<td>sterile for surgery</td>
		</tr>
		<tr>
			<td>Skin biopsy punches</td>
			<td></td>
			<td></td>
			<td>Size as determined by researcher</td>
		</tr>
		<tr>
			<td>Slide boxes</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Slide warmers</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Superfrosted microscope slides</td>
			<td>Fisher Scientific</td>
			<td>22 037 246</td>
			<td></td>
		</tr>
		<tr>
			<td>Temperature control water bath</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Tissue embedding station</td>
			<td></td>
			<td></td>
			<td>Minimum of a paraffin dispenser and a cold plate</td>
		</tr>
		<tr>
			<td>Tissue processor</td>
			<td></td>
			<td></td>
			<td>Minimum of a oven with a vacuum pump</td>
		</tr>
		<tr>
			<td>Triple antibiotic opthalmic ointment</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>tweezers, curved tip</td>
			<td></td>
			<td></td>
			<td>sterile for surgery</td>
		</tr>
		<tr>
			<td>tweezers, tapered tip</td>
			<td></td>
			<td></td>
			<td>sterile for surgery</td>
		</tr>
		<tr>
			<td>WypAll X60</td>
			<td>Kimberly-Clark</td>
			<td>34865</td>
			<td></td>
		</tr>
	</tbody>
</table>

murine excisional wound healing model and histological morphometric wound analysis

穆林切除伤口模型已被广泛使用，用于研究伤口愈合的每个连续重叠阶段：炎症，增殖和重塑。Murine 伤口具有组织学上定义明确且易于识别的伤口床，这些不同阶段的愈合过程是可测量的。在该领域，通常使用任意定义的伤口"中间"进行组织学分析。然而，伤口是一个三维实体，往往不是组织对称的，支持需要一个定义明确和稳健的定量方法，以检测小效果尺寸的形态缺陷。在该协议中，我们描述了在小鼠中创建双边全厚度切除伤口的过程，以及有关如何使用特定序列部分的图像处理程序测量形态参数的详细说明。伤口长度、表皮长度、表皮面积和伤口面积的两维测量与各部之间的已知距离相结合，以推断覆盖伤口、整体伤口面积、表皮体积和伤口体积的三维表皮面积。虽然这种详细的组织学分析比常规分析更耗时和资源，但其严谨性增加了在固有的复杂伤口愈合过程中检测新表型的可能性。

图5描述了通过对多个外科医生在不同小鼠菌株中产生的野生型伤口进行形态分析，并由不同个体进行分析，获得的测量和计算值的范围。不同菌株的野生小鼠可以显示统计差异，如我们的研究和文献9，10所述。根据这些具有代表性的结果，我们建议在一项研究中，只使用一种菌株的小鼠。虽然我们建议同一个人在特定研究中执行?...

Watch this Scientific Journal Video about Murine Excisional Wound Healing Model and Histological Morphometric Wound Analysis at JoVE.com

Murine Excisional Wound Healing Model and Histological Morphometric Wound Analysis

该协议描述了如何在小鼠中生成双边全厚度切除伤口，以及如何随后监测、收获和准备伤口以进行形态分析。其中包括一个深入的描述，说明如何使用串行组织学部分来定义、精确量化和检测形态缺陷。

穆林切除伤口愈合模型与组织学形态性伤口分析

The murine excisional wound model has been used extensively to study each of the sequentially overlapping phases of wound healing: inflammation, ...

该协议使用组织学分析，以获得对小鼠切除伤口的强健形态测量。使用这种技术，我们可以使用串行部分的子集来分析整个伤口，使我们能够更准确地检测可能遗漏的缺陷。展示这个程序的将是林赛·瑞亚，一个来自我实验室的副研究员。在确认麻醉成年小鼠对踏板反射缺乏反应后，对动物的眼睛涂抹软膏，并在肩-肩形运动中使用电动剃须刀剪子，以去除肩部小鼠背部的毛皮。使用剃须刀刀片从背部 20 度角在旋转-肌体运动中去除暴露的皮肤的任何剩余头发，然后用一个波维酮碘和一个 70%异丙醇准备垫擦拭清洁皮肤。对于伤口感应，将干净的纸质毛巾披在覆盖着牙蜡的平坦表面上，然后将鼠标放在毛巾的易发位置。沿着后心中线在肩部刀片之间捏住鼠标的皮肤，将夹紧的皮肤从身体上拉开。将鼠标切换到侧卧位置，将所需伤口大小的活检冲头尽可能靠近身体，让皮肤放松。使用摇摆动作按下冲孔，刺穿皮肤的所有层，并使用无菌剪刀和钳子从伤口上去除冲孔活检，以必要时从周围皮肤上释放冲孔。然后让鼠标通过监测和镇痛恢复。对于伤口床的形态分析，打开数字染色伤口图像。要设置比例和测量首选项，请在"分析"下选择"设置比例"并输入以像素为单位的距离、已知距离和长度单位。选择"全局"和"确定"可使每个打开的图像的缩放保持不变，然后选择"分析"和"设置测量"以确认已选中区域框。要测量伤口长度，请选择徒手，并使用工具沿皮肤-表皮结段进行跟踪，从伤口一侧未受伤组织的最后一个毛囊开始，到另一侧未受伤组织的第一个毛囊。如果表皮不覆盖整个伤口，跟随伤口一侧的皮皮-表皮结，迁移舌头结束，并继续遵循造粒组织或造粒组织与鳞片之间的交汇点，直到到达伤口另一侧未受伤组织的迁移舌头和未受伤组织的第一个毛囊。然后单击"分析和测量"。测量的长度将显示为与刻度中设置的相同单位。如果表皮未覆盖整个伤口，则根据造粒组织优越的方面或造粒组织与鳞片之间的交汇点测量每个表皮前缘之间的距离。要测量伤口区域，请使用徒手工具沿表皮的优越方面或造粒组织的优越方面进行跟踪，从伤口一侧未受伤的组织的最后一个毛囊开始，到另一侧未受伤的组织的第一个毛囊。继续沿着毛囊垂直追踪到造粒组织。一旦到达相反的毛囊和脂肪组织或肌肉，沿着造粒组织的劣边界到伤口的对面，并沿毛囊连接起点以关闭该区域。如果伤口未完全上皮化，沿表皮的优越方面进行跟踪，直到前缘，然后返回到皮肤-表皮结后的起点。如图所示，在整个伤口进行二D测量时，我们能够仅对伤口中间进行二D分析，从而计算出不可能计算的参数，这是现场常用的方法。在这个实验中，对伤口面积进行了二D测量，对整个伤口或伤口的中间子集进行了平均测量，并未发现实验组或分析方法之间的显著差异。然而，计算出的伤口体积在实验组之间有显著差异，这证明了对伤口愈合参数进行深入组织学分析的重要性。为组织学分析生成一致大小的伤口并执行严格的串行剖截，对于精确的形态分析至关重要。未染色的部分可用于免疫荧光或马森的三色染色，以测量伤口愈合的其他方面，而闪光冷冻伤口可用于形态测定，用于定量蛋白质分析。

Research

JoVE Journal

Developmental Biology

Zebrafish Keratocyte Explants to Study Collective Cell Migration and Reepithelialization in Cutaneous Wound Healing

斑马鱼角膜植研究集体细胞迁移和再上皮化在皮肤伤口愈合

Due to their unique motile properties, fish keratocytes dissociated from explant cultures have long been used to study the mechanisms of single cell ...

科研

发育生物学

Automated Quantification of Hematopoietic Cell &#8211; Stromal Cell Interactions in Histological Images of Undecalcified Bone

造血干细胞的自动定量 - 在脱钙骨的组织学图像间质细胞的相互作用

Confocal microscopy is the method of choice for the analysis of localization of multiple cell types within complex tissues such as the bone marrow. ...

Establishment of a Clinically Relevant Ex Vivo Mock Cataract Surgery Model for Investigating Epithelial Wound Repair in a Native Microenvironment

Establishment of a Clinically Relevant Ex Vivo Mock Cataract Surgery Model for Investigating Epithelial Wound Repair in a Native Microenvironment

建立临床相关<em&gt;离体</em&gt;调查上皮创伤修复的微环境本地人模拟白内障手术模型

The major impediment to understanding how an epithelial tissue executes wound repair is the limited availability of models in which it is possible to ...

Optimization of the Wound Scratch Assay to Detect Changes in Murine Mesenchymal Stromal Cell Migration After Damage by Soluble Cigarette Smoke Extract

伤口划痕试验的优化来探测在小鼠间充质干细胞迁移的可溶性香烟烟雾提取物受损后

Cell migration is vital to many physiological and pathological processes including tissue development, repair, and regeneration, cancer metastasis, and ...

Adenoviral Gene Therapy for Diabetic Keratopathy: Effects on Wound Healing and Stem Cell Marker Expression in Human Organ-cultured Corneas and Limbal Epithelial Cells

腺病毒基因治疗对糖尿病性角膜病变：对伤口愈合和干细胞标记表达的人体器官培养角膜和角膜缘上皮细胞

The goal of this protocol is to describe molecular alterations in human diabetic corneas and demonstrate how they can be alleviated by adenoviral gene ...

Culture of Murine Embryonic Metatarsals: A Physiological Model of Endochondral Ossification

小鼠胚胎跖骨文化：软骨内骨化的生理模型

The fundamental process of endochondral ossification is under tight regulation in the healthy individual so as to prevent disturbed development and/or ...

Rapid Isolation of BMPR-IB+ Adipose-Derived Stromal Cells for Use in a Calvarial Defect Healing Model

BMPR-IB +快速分离脂肪来源的使用基质细胞在颅骨缺损愈合模型

Invasive cancers, major injuries, and infection can cause bone defects that are too large to be reconstructed with preexisting bone from the patient's own ...

A Device for Performing Cell Migration/Wound Healing in a 96-Well Plate

一种在96孔板表演细胞迁移/伤口愈合设备

The cell migration/wounding assay is a commonly used method to study cell migration and other biological processes, such as angiogenesis and tumor ...

Histological Analyses of Acute Alcoholic Liver Injury in Zebrafish

斑马鱼急性酒精性肝损伤的组织学分析

Alcoholic Liver Disease (ALD) refers to damage to the liver due to acute or chronic alcohol abuse. It is among the leading causes of alcohol-related ...

Analysis of Protein-protein Interactions and Co-localization Between Components of Gap, Tight, and Adherens Junctions in Murine Mammary Glands

蛋白质 - 蛋白质相互作用的分析和鼠乳腺中间隙，紧密和粘附连接部位之间的共定位

Cell-cell interactions play a pivotal role in preserving tissue integrity and the barrier between the different compartments of the mammary gland. These ...