Name: visualizing scar development using scad assay - an ex-situ skin scarring assay
Uploaded: 2022-04-28T13:40:00
Description: Watch this Scientific Journal Video about Visualizing Scar Development Using SCAD Assay - An Ex-situ Skin Scarring Assay at JoVE.com

我们感谢Jiang et al. 2020的所有合著者为SCAD方法13的发展做出了贡献。我们感谢Steffen Dietzel博士和路德维希- 马克西米兰斯大学生物成像核心设施对多光子系统的访问。Y.R.得到了Else-Kröner-Fresenius-Stiftung（2016_A21），欧洲研究理事会整合基金（ERC-CoG 819933）和LEO基金会（LF-OC-21-000835）的支持。

下面介绍的模型提供了SCAD测定生成的详细分步描述，如Jiang等人，202013中简要描述的那样。SCAD样品制备是在根据国际和上巴伐利亚政府的指导方针牺牲动物后进行的。动物被安置在慕尼黑亥姆霍兹中心的动物设施中。房间保持在最佳湿度和恒定温度下，光照周期为12小时。动物们 被随意地提供食物和水。
1. 全身心塌病组织制备
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<ol>
	<li>Longaker, M. 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=Adult+skin+wounds+in+the+fetal+environment+heal+with+scar+formation.">Adult skin wounds in the fetal environment heal with scar formation.</a> Annals of Surgery. 219 (1), 65-72 (1994).</li><li>desJardins-Park, H. E., Foster, D. S., 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=Fibroblasts+and+wound+healing:+an+update.">Fibroblasts and wound healing: an update.</a> Regenerative Medicine. 13 (5), 491-495 (2018).</li><li>Jiang, X., Iseki, S., Maxson, R. E., Sucov, H. M., Morriss-Kay, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tissue+origins+and+interactions+in+the+mammalian+skull+vault.">Tissue origins and interactions in the mammalian skull vault.</a> Developmental Biology. 241 (1), 106-116 (2002).</li><li>Tripathi, 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=Hypertrophic+scars+and+keloids:+a+review+and+current+treatment+modalities.">Hypertrophic scars and keloids: a review and current treatment modalities.</a> Biomedical Dermatology. 4, 11 (2020).</li><li>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+healing--Aiming+for+perfect+skin+regeneration.">Wound healing--Aiming for perfect skin regeneration.</a> Science. 276 (5309), 75-81 (1997).</li><li>Correa-Gallegos, 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=Patch+repair+of+deep+wounds+by+mobilized+fascia.">Patch repair of deep wounds by mobilized fascia.</a> Nature. 576 (7786), 287-292 (2019).</li><li>Sen, C. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+wounds+and+its+burden:+An+updated+compendium+of+estimates.">Human wounds and its burden: An updated compendium of estimates.</a> Advances in Wound Care. 8 (2), 39-48 (2019).</li><li>Rinkevich, 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=Identification+and+isolation+of+a+dermal+lineage+with+intrinsic+fibrogenic+potential.">Identification and isolation of a dermal lineage with intrinsic fibrogenic potential.</a> Science. 348 (6232), 2151 (2015).</li><li>Leavitt, 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=Prrx1+fibroblasts+represent+a+pro-fibrotic+lineage+in+the+mouse+ventral+dermis.">Prrx1 fibroblasts represent a pro-fibrotic lineage in the mouse ventral dermis.</a> Cell Reports. 33 (6), 108356 (2020).</li><li>Driskell, R. 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=Distinct+fibroblast+lineages+determine+dermal+architecture+in+skin+development+and+repair.">Distinct fibroblast lineages determine dermal architecture in skin development and repair.</a> Nature. 504 (7479), 277-281 (2013).</li><li>Walmsley, G. 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=Live+fibroblast+harvest+reveals+surface+marker+shift+in+vitro.">Live fibroblast harvest reveals surface marker shift in vitro.</a> Tissue Engineering. Part C, Methods. 21 (3), 314-321 (2015).</li><li>Hakkinen, K. M., Harunaga, J. S., Doyle, A. D., Yamada, K. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+comparisons+of+the+morphology,+migration,+cell+adhesions,+and+actin+cytoskeleton+of+fibroblasts+in+four+different+three-dimensional+extracellular+matrices.">Direct comparisons of the morphology, migration, cell adhesions, and actin cytoskeleton of fibroblasts in four different three-dimensional extracellular matrices.</a> Tissue Engineering. Part A. 17 (5-6), 713-724 (2011).</li><li>Jiang, 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=Injury+triggers+fascia+fibroblast+collective+cell+migration+to+drive+scar+formation+through+N-cadherin.">Injury triggers fascia fibroblast collective cell migration to drive scar formation through N-cadherin.</a> Nature Communications. 11 (1), 5653 (2020).</li><li>Wan, 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=Connexin43+gap+junction+drives+fascia+mobilization+and+repair+of+deep+skin+wounds.">Connexin43 gap junction drives fascia mobilization and repair of deep skin wounds.</a> Matrix Biology: Journal of the International Society for Matrix Biology. 97, 58-71 (2021).</li><li>Molbay, M., Kolabas, Z. I., Todorov, M. I., Ohn, T. -. L., Ert&#252;rk, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+guidebook+for+DISCO+tissue+clearing.">A guidebook for DISCO tissue clearing.</a> Molecular Systems Biology. 17 (3), 9807 (2021).</li><li>Ueda, H. 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=Tissue+clearing+and+its+applications+in+neuroscience.">Tissue clearing and its applications in neuroscience.</a> Nature Reviews Neuroscience. 21 (2), 61-79 (2020).</li><li>Ert&#252;rk, 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=Three-dimensional+imaging+of+solvent-cleared+organs+using+3DISCO.">Three-dimensional imaging of solvent-cleared organs using 3DISCO.</a> Nature Protocols. 7 (11), 1983-1995 (2012).</li><li>Wilhelm, K. -. P., Wilhelm, D., Bielfeldt, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Models+of+wound+healing:+an+emphasis+on+clinical+studies.">Models of wound healing: an emphasis on clinical studies.</a> Skin Research and Technology: Official Journal of International Society for Bioengineering and the Skin (ISBS) [and] International Society for Digital Imaging of Skin (ISDIS) [and] International Society for Skin Imaging (ISSI). 23 (1), 3-12 (2017).</li><li>Grada, A., Mervis, J., Falanga, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Research+techniques+made+simple:+Animal+models+of+wound+healing).">Research techniques made simple: Animal models of wound healing).</a> The Journal of Investigative Dermatology. 138 (10), 2095-2105 (2018).</li></ol>

已经开发了几种模型来了解受伤后疤痕的形成。虽然在这方面取得了许多进展，但实际机制仍不清楚。与以前的技术相比，SCAD模型包含所有细胞类型和皮肤层，从而保持了天然皮肤18，19的复杂性。这种方法能够生成基本数据集，这些数据集对于理解驱动疤痕形成的分子机制非常重要。该测定以简单，简约的方式设计，但能够产生基本和功能读数，这对...

伤口愈合是恢复破裂伤口的过程。无脊椎动物的组织损伤导致部分或完全再生。相比之下，哺乳动物通过疤痕形成对深部损伤做出反应，这是一种量身定制的过程，用于用密集的基质纤维塞快速密封伤口，从而最大限度地减少破坏区域，同时使受伤部位永久变形1，2，3。哺乳动物的大面积皮肤灼伤或深部开放性伤口导致病理性表型，例如肥厚性或瘢痕疙瘩4，5。这些茂密的疤痕给临床和全球医疗保健系统带来了巨大的负担。仅在美国，疤痕管理每年的成本约为100亿美元，6，7。因此，需要开发相关方法，以更好地了解疤痕形成所涉及的基础过程和机制。
近年来，对小鼠的广泛研究揭示了异质成纤维细胞群体，这些成纤维细胞群体基于它们在某些皮肤位置的起源具有不同的功能效力8，9，10。在后皮中，Rinkevich等人，2015年，发现具有Engrailed-1（En1）早期胚胎表达的特定成纤维细胞群体，称为EPF（Engrailed阳性成纤维细胞）有助于受伤时的皮肤瘢痕形成。相反，另一种没有受累表达史的成纤维细胞谱系，Engrailed阴性成纤维细胞（ENF），不会导致瘢痕形成8。这些En1谱系的命运图谱使用Cre驱动的转基因小鼠品系交叉到荧光报告小鼠品系，如R26mTmG （En1Cre x R26mTmG），可以可视化EPF和ENF种群。
在几天 内 研究体内成纤维细胞迁移受到伦理和技术限制的限制。此外，化合物、病毒和中和抗体库筛选以调节瘢痕形成所涉及的途径在技术上具有挑战性。以前使用的 体外 或 离体 模型缺乏在真实皮肤微环境中可视化成纤维细胞迁移和疤痕形成的能力，疤痕发育的均匀性，以及模拟 体内 皮肤环境的组织复杂性11，12。为了克服上述局限性，我们开发了一种称为SCAD（A培养皿中的SCar样组织）的 离体 瘢痕测定法13，14。这种简单的测定可以通过切除含有表皮，真皮和皮下筋膜区域的2mm全层皮肤并在血清补充的DMSO培养基中培养长达5天来进行。SCAD产生的瘢痕可靠地复制 体内 瘢痕的转录组和蛋白质组学特征。此外，从相关转基因小鼠品系（例如En1小鼠）与荧光报告小鼠品系杂交产生的SCAD能够以前所未有的分辨率可视化成纤维细胞迁移动力学和疤痕发展。此外，该模型可以很容易地适应任何高通量应用（例如，化合物库、抗体库或病毒筛选）13，14。在本文中，我们描述了一种优化的方案，用于生成SCAD和随后的下游处理应用，以研究疤痕发育中的细胞和基质动力学。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>10% Tween 20, Nonionic Detergent</td>
			<td>Biorad Laboratories</td>
			<td>1610781</td>
			<td></td>
		</tr>
		<tr>
			<td>Bovine serum albumin, Cold ethanol fract</td>
			<td>Sigma</td>
			<td>A4503-50G</td>
			<td></td>
		</tr>
		<tr>
			<td>DMEM/F-12, HEPES, no phenol red-500 mL</td>
			<td>LIFE Technologies</td>
			<td>11039021</td>
			<td></td>
		</tr>
		<tr>
			<td>DPBS, no calcium, no magnesium</td>
			<td>Gibco</td>
			<td>14190169</td>
			<td></td>
		</tr>
		<tr>
			<td>Epredia Cryostar NX70 Cryostat</td>
			<td>Thermo Scientific</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Epredia SuperFrost Plus Adhesion slides</td>
			<td>Fisher scientific</td>
			<td>J1800AMNZ</td>
			<td>Adhesion slides</td>
		</tr>
		<tr>
			<td>Fetal Bovine Serum, qualified, heat inactivated, E.U.-approved, South America Origin-500 mL</td>
			<td>LIFE Technologies</td>
			<td>&#160;10500064</td>
			<td></td>
		</tr>
		<tr>
			<td>Fluoromount-G with DAPI</td>
			<td>Life Technologies</td>
			<td>00 4959 52</td>
			<td>Mounting medium with DAPI</td>
		</tr>
		<tr>
			<td>Forceps curved with fine points with guidepinstainless steel(tweezers)125 mm length</td>
			<td>Fisher Scientific</td>
			<td>12381369</td>
			<td></td>
		</tr>
		<tr>
			<td>Gelatin from porcine skin</td>
			<td>Sigma</td>
			<td>G2500-100G</td>
			<td></td>
		</tr>
		<tr>
			<td>GlutaMAX Supplement-100 mL</td>
			<td>LIFE Technologies</td>
			<td>35050038</td>
			<td></td>
		</tr>
		<tr>
			<td>HBSS, calcium, magnesium, no phenol red-500 mL</td>
			<td>LIFE Technologies</td>
			<td>14025092</td>
			<td></td>
		</tr>
		<tr>
			<td>Ibidi Gas incubation system for CO2 and O2</td>
			<td>Ibidi</td>
			<td>11922</td>
			<td></td>
		</tr>
		<tr>
			<td>Ibidi Heating system</td>
			<td>Ibidi</td>
			<td>10915</td>
			<td></td>
		</tr>
		<tr>
			<td>Leica SP8 upright microscope - Multiphoton excitation 680&#8211;1300 nm</td>
			<td>Leica</td>
			<td></td>
			<td>Equipped with a 25x water-dipping objective (HC IRAPO L 25x/1.00&#8201;W) in combination with a tunable laser (Spectra-Physics, InSight DS&#8201;+&#8201;Single)</td>
		</tr>
		<tr>
			<td>Non Essential Amino Acids</td>
			<td>LIFE Technologies</td>
			<td>11140035</td>
			<td></td>
		</tr>
		<tr>
			<td>NuSieve GTG Agarose ,25 g</td>
			<td>Biozym /Lonza</td>
			<td>859081</td>
			<td></td>
		</tr>
		<tr>
			<td>OCT Embedding Matrix</td>
			<td>Carlroth</td>
			<td>6478.1</td>
			<td></td>
		</tr>
		<tr>
			<td>Paraformaldehyde, 16% W/V AQ. 10 x10 mL</td>
			<td>VWR International</td>
			<td>43368.9M</td>
			<td></td>
		</tr>
		<tr>
			<td>Pen-Strep</td>
			<td>Gibco</td>
			<td>15140122</td>
			<td></td>
		</tr>
		<tr>
			<td>Stiefel Biopsy-Punch 2 mm</td>
			<td>Stiefel</td>
			<td>270130</td>
			<td></td>
		</tr>
		<tr>
			<td>Straight Sharp/Sharp Dissecting Scissors 11.4 cm</td>
			<td>Fisher Scientific</td>
			<td>15654444</td>
			<td></td>
		</tr>
		<tr>
			<td>Thimerosal Bioxtra, 97%&#8211;101%</td>
			<td>Sigma-Aldrich</td>
			<td>T8784-1G</td>
			<td></td>
		</tr>
		<tr>
			<td>Zeiss Axioimager M2 upright microscope</td>
			<td>Zeiss</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

visualizing scar development using scad assay - an ex-situ skin scarring assay

哺乳动物对密封深部组织伤口的整体反应是通过瘢痕形成和组织收缩，由专门的筋膜成纤维细胞介导。尽管疤痕形成和伤口愈合受损具有临床意义，但由于缺乏相关的测定，我们可以直接可视化成纤维细胞在复杂环境（如皮肤伤口）中的成纤维细胞编导和动力学，因此我们对伤口愈合中筋膜成纤维细胞动力学的理解是粗略的。本文提出了一种使用SCAD或“培养皿中的SCar样组织”来模拟皮肤伤口的复杂环境来生成 异地 皮肤疤痕的方案。在该测定中，切除2mm全厚皮肤并在培养基中倒置培养5天，在此期间疤痕和皮肤挛缩均匀发展。这种方法与成纤维细胞谱系特异性转基因小鼠模型相结合，可以在整个伤口修复过程中可视化单个成纤维细胞谱系。总体而言，该协议有助于研究人员了解伤口修复的基本过程和机制，直接探索调节剂对伤口愈合结果的影响。

SCAD的产生可以分为三个基本步骤：从P0-P1小鼠中收集背部皮肤，产生全层活检冲孔，随后在96孔板中培养单个粪便长达5天。作为读数，该测定可以进一步应用于分析瘢痕形成的空间和时间方面。空间分析利用组织的2D和3D免疫标记来研究发育中的疤痕组织中细胞和基质成分的空间定位。时空研究允许使用组织内在或外在荧光蛋白来可视化这些 非原位 疤痕，这些荧光蛋白可以使用相关的3D延?...

Watch this Scientific Journal Video about Visualizing Scar Development Using SCAD Assay - An Ex-situ Skin Scarring Assay at JoVE.com

Visualizing Scar Development Using SCAD Assay - An Ex-situ Skin Scarring Assay

该协议描述了称为“培养皿中的SCar样组织”或SCAD的皮肤筋膜外植体的产生。该模型允许在疤痕形成过程中对单个成纤维细胞进行前所未有的可视化。

使用SCAD测定可视化疤痕形成 - 一种 非原位 皮肤疤痕测定

The mammalian global response to sealing deep tissue wounds is through scar formation and tissue contraction, mediated by specialized fascia fibroblasts. ...

以前使用的体外或体外模型缺乏真皮细胞成分和皮肤组织的复杂性。这种非原位SCAD测定克服了一定的局限性，并能够研究受伤动物外部疤痕的发展。SCAD测定可以可视化皮肤微环境中的成纤维细胞迁移和疤痕形成。它使激活剂或抑制剂的筛选库能够了解疤痕形成的机制基础。使用SCAD模型对平面文库进行高通量筛选分析，以了解伤口修复中涉及的基本过程和分子过程。SCAD测定很容易使用皮肤外植体进行。对于一致的疤痕，建议选择产后第零天或第一天的皮肤。牺牲产后零日或第一天新生幼犬后，使用无菌手术刀小心切除1.5×1.5厘米全厚的背背皮肤，直到骨骼肌层。使用无菌弯曲镊子剥离皮肤，确保浅筋膜与下面的全皮肌完整。用50至100毫升冷DMEM F-12培养基清洗切除的组织，以除去污染的血液。然后，用Hanks平衡盐溶液洗涤组织，以保持组织和细胞活力。将皮肤倒置，顶部有浅表筋膜，放入含有DMEM F-12培养基的10厘米培养皿中。然后，使用一次性两毫米活检冲头，切除全层圆形皮肤碎片以产生结痂组织，确保浅筋膜完好无损，下层掌骨肌直到表皮。将200微升不含酚红的新鲜DMEM F-12完整培养基加入96孔板的每个孔中。使用无菌镊子，将单个结痂组织倒置并完全浸没到96孔板的孔中。将板转移到维持在标准条件下的细胞培养箱中。在培养的第二天和第四天，除去在孔中留下10微升的培养基，并加入新鲜预热的DMEM F-12完整培养基以及处理化合物，以维持持续的细胞和组织活力条件。对于SCAD的实时成像，通过在微波炉中加热，在玻璃瓶中制备30毫升2至3%低熔点琼脂糖溶液PBS。煮沸后，立即转移瓶子并将液体琼脂糖溶液在40摄氏度的水浴中冷却。接下来，将筋膜或疤痕朝上转移到35毫米培养皿的中心的SCAD组织上。然后使用1000微升移液器吸头将40摄氏度的琼脂糖液体缓慢倒入组织中，在室温下包埋SCAD。琼脂糖在两分钟内聚合。聚合后，加入两毫升预热的DMEM F-12完整培养基。然后使用共聚焦或配备手稿中解释的合适孵育系统的多光子显微镜，获取第零天至第一天的延时图像。对于组织收获，通过在相关时间点用无菌PBS替换培养基来洗涤组织。使用无菌镊子将每个SCAD转移到含有500微升2%多聚甲醛的1.5毫升微量离心管中，以在4摄氏度下固定组织过夜。第二天，用PBS洗涤组织三次，然后按照文本手稿中的描述进行二维或三维免疫荧光染色。此处显示了 SCAD 在第 0 天和第 5 天的代表性全安装明场图像。Masson对垂直组织切片的三色染色揭示了组织瘢痕形成，组织收缩以及第0天和第5天瘢痕核心细胞外基质积聚的特征。显示了SCADs在第零天和第五天的免疫荧光染色。从早期和晚期瘢痕，根深蒂固的阳性成纤维细胞显示为绿色，根深蒂固的阴性成纤维细胞显示为红色。对于三维成像，将组织嵌入琼脂糖溶液中并在上面加PBSGT。具有代表性的图像展示了N-钙粘蛋白在第五天SCAD的疤痕部位的独家定位。配备孵育室的三维延时成像装置显示了成纤维细胞群在前12小时或疤痕发育的早期阶段进展的早期事件。这里给出了单个成纤维细胞在疤痕发育过程中单细胞跟踪细胞轨迹的图形表示。将SCAD组织倒置在孔板内以使筋膜朝上至关重要。如果不这样做，将导致迁移模式不可避免地发生变化。该程序允许使用化学调节剂，中和抗体或病毒方法检查真皮层以进行治疗或培养。这有助于评估不同医疗环境中的病理性纤维化反应。我们展示了连接蛋白-43和N-钙粘蛋白作为损伤时参与筋膜动员的关键分子的表达。SCAD方法允许确定新的治疗策略来减少瘢痕形成和纤维化。

Research

JoVE Journal

Biology

Visualizing Scar Development Using SCAD Assay - An Ex-situ Skin Scarring Assay

Titration of Human Coronaviruses Using an Immunoperoxidase Assay

人类冠状病毒的滴定使用免疫过氧化物酶含量

Determination of infectious viral titers is a basic and essential experimental approach for virologists. Classical plaque assays cannot be used for ...

科研

生物学

A Neuronal and Astrocyte Co-Culture Assay for High Content Analysis of Neurotoxicity

一个神经元和星形胶质细胞共培养检测神经毒性的高含量分析

High Content Analysis (HCA) assays combine cells and detection reagents with automated imaging and powerful image analysis algorithms, allowing ...

Visualizing the Beating Heart in Drosophila

Visualizing the Beating Heart in Drosophila

可视化心脏跳动果蝇</em

The Drosophila heart has recently emerged as a good model system for examining the genetic, cellular, and molecular mechanisms underlying function in ...

An In vitro FluoroBlok Tumor Invasion Assay

An In vitro FluoroBlok Tumor Invasion Assay

一个<em在体外 FluoroBlok肿瘤侵袭实验

The hallmark of metastatic cells is their ability to invade through the basement membrane and migrate to other parts of the body. Cells must be able to ...

Conversion of a Capture ELISA to a Luminex xMAP Assay using a Multiplex Antibody Screening Method

转换到一个点阵仪XMAP含量，使用多重抗体筛查方法的捕获ELISA

The enzyme-linked immunosorbent assay (ELISA) has long been the primary tool for detection of analytes of interest in biological samples for both life ...

Analysis of SNARE-mediated Membrane Fusion Using an Enzymatic Cell Fusion Assay

SNARE蛋白介导的膜融合酶的细胞融合含量的分析

The interactions of SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor)  proteins on vesicles (v-SNAREs) and on target membranes ...

Measuring the Effects of Bacteria on C. Elegans Behavior Using an Egg Retention Assay

Measuring the Effects of Bacteria on C. Elegans Behavior Using an Egg Retention Assay

测量细菌的影响<em&gt; C。线虫</em&gt;行为使用一个鸡蛋保留含量

C. elegans egg-laying behavior is  affected by environmental cues such as osmolarity1 and  vibration2. In the total absence of food C. elegans also cease ...

Development of a Quantitative Recombinase Polymerase Amplification Assay with an Internal Positive Control

一个定量聚合酶重组酶扩增法的发展与内部阳性对照

It was recently demonstrated that recombinase polymerase amplification (RPA), an isothermal amplification platform for pathogen detection, may be used to ...

G Protein-selective GPCR Conformations Measured Using FRET Sensors in a Live Cell Suspension Fluorometer Assay

在活细胞悬浮液荧光测定G蛋白选择性GPCR构象测量使用FRET传感器

F&#1255;rster resonance energy transfer (FRET)-based studies have become increasingly common in the investigation of GPCR signaling. Our research group ...

Evaluating Vascular Hyperpermeability-inducing Agents in the Skin with the Miles Assay

用迈尔斯测定法评价皮肤血管 Hyperpermeability 诱导剂的含量

The primary function of the vascular endothelium in vertebrate organisms is to serve as a barrier between the blood and each tissue of the body, whereby ...

使用SCAD测定可视化疤痕形成 - 一种 非原位 皮肤疤痕测定

使用SCAD测定可视化疤痕形成 - 一种非原位皮肤疤痕测定