Name: whole-mount confocal microscopy for adult ear skin: a model system to study neuro-vascular branching morphogenesis and immune cell distribution
Uploaded: 2018-03-29T15:00:00
Description: Watch this Scientific Journal Video about Whole-mount Confocal Microscopy for Adult Ear Skin: A Model System to Study Neuro-vascular Branching Morphogenesis and Immune Cell Distribution at JoVE.com

我们感谢吉尔的实验室管理和技术支持, j. 霍金斯和国家卫生研究院 (NIH) 的工作人员50建立了援助老鼠护理的动物设施, 以及里德和 f. Baldrey 的行政援助。还感谢 s Motegi 和 m. Udey 分享他们的耳朵皮肤解剖协议, n. 烧伤的编辑帮助, 以及干细胞和神经血管生物学实验室成员的技术帮助和深思熟虑的讨论。KAITOKU 是日本促进科学协会 (jsp) 的支持。这项工作得到了国家心脏、肺和血液研究所 (HL005702-11 到 Y.M.) 的校内研究计划的支持。

这一节的所有实验都是在国家心脏、肺和血液研究所 (NHLBI) 动物护理和使用委员会的批准下进行的。
1. 成人小鼠耳皮收集
<ol><li>弄死成年小鼠的二氧化碳 (CO2) 暴露在一个封闭的房间, 然后确认安乐死的颈椎脱位。 注: 实验遵循国家卫生研究院 (NIH) 的安乐死方法指南。</li>
	<li>从底座解剖耳朵, 将其放置在 35 x 10 毫米的2培养皿中, 其中包含有2毫升的汉克平?...

<ol>
	<li>Mukouyama, Y. S., Shin, D., Britsch, S., Taniguchi, M., Anderson, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sensory+nerves+determine+the+pattern+of+arterial+differentiation+and+blood+vessel+branching+in+the+skin.">Sensory nerves determine the pattern of arterial differentiation and blood vessel branching in the skin.</a> Cell. 109, 693-705 (2002).</li><li>Mukouyama, Y. S., James, J., Nam, J., Uchida, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Whole-mount+confocal+microscopy+for+vascular+branching+morphogenesis.">Whole-mount confocal microscopy for vascular branching morphogenesis.</a> Methods Mol Biol. 843, 69-78 (2012).</li><li>Li, W., Mukouyama, Y. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Whole-mount+immunohistochemical+analysis+for+embryonic+limb+skin+vasculature:+a+model+system+to+study+vascular+branching+morphogenesis+in+embryo.">Whole-mount immunohistochemical analysis for embryonic limb skin vasculature: a model system to study vascular branching morphogenesis in embryo.</a> J Vis Exp. , (2011).</li><li>Yamazaki, 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=Tissue+Myeloid+Progenitors+Differentiate+into+Pericytes+through+TGF-beta+Signaling+in+Developing+Skin+Vasculature.">Tissue Myeloid Progenitors Differentiate into Pericytes through TGF-beta Signaling in Developing Skin Vasculature.</a> Cell Rep. 18, 2991-3004 (2017).</li><li>Mukouyama, Y. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vessel-dependent+recruitment+of+sympathetic+axons:+looking+for+innervation+in+all+the+right+places.">Vessel-dependent recruitment of sympathetic axons: looking for innervation in all the right places.</a> J Clin Invest. 124, 2855-2857 (2014).</li><li>Li, W., 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=Peripheral+nerve-derived+CXCL12+and+VEGF-A+regulate+the+patterning+of+arterial+vessel+branching+in+developing+limb+skin.">Peripheral nerve-derived CXCL12 and VEGF-A regulate the patterning of arterial vessel branching in developing limb skin.</a> Dev Cell. 24, 359-371 (2013).</li><li>Chang, H., Wang, Y., Wu, H., Nathans, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Flat+mount+imaging+of+mouse+skin+and+its+application+to+the+analysis+of+hair+follicle+patterning+and+sensory+axon+morphology.">Flat mount imaging of mouse skin and its application to the analysis of hair follicle patterning and sensory axon morphology.</a> J Vis Exp. , e51749 (2014).</li><li>Salz, L., Driskell, R. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Horizontal+Whole+Mount:+A+Novel+Processing+and+Imaging+Protocol+for+Thick,+Three-dimensional+Tissue+Cross-sections+of+Skin.">Horizontal Whole Mount: A Novel Processing and Imaging Protocol for Thick, Three-dimensional Tissue Cross-sections of Skin.</a> J Vis Exp. , (2017).</li><li>Liakath-Ali, 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=Novel+skin+phenotypes+revealed+by+a+genome-wide+mouse+reverse+genetic+screen.">Novel skin phenotypes revealed by a genome-wide mouse reverse genetic screen.</a> Nat Commun. 5, 3540 (2014).</li><li>Gunawan, 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=The+methyltransferase+Ezh2+controls+cell+adhesion+and+migration+through+direct+methylation+of+the+extranuclear+regulatory+protein+talin.">The methyltransferase Ezh2 controls cell adhesion and migration through direct methylation of the extranuclear regulatory protein talin.</a> Nat Immunol. 16, 505-516 (2015).</li><li>Avci, 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=Animal+models+of+skin+disease+for+drug+discovery.">Animal models of skin disease for drug discovery.</a> Expert Opin Drug Dis. 8, 331-355 (2013).</li><li>Jin, H., He, R., Oyoshi, M., Geha, R. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+of+atopic+dermatitis.">Animal models of atopic dermatitis.</a> J Invest Dermatol. 129, 31-40 (2009).</li><li>Wagner, E. F., Schonthaler, H. B., Guinea-Viniegra, J., Tschachler, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Psoriasis:+what+we+have+learned+from+mouse+models.">Psoriasis: what we have learned from mouse models.</a> Nat Rev Rheumatol. 6, 704-714 (2010).</li><li>Nunan, R., Harding, K. G., 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=Clinical+challenges+of+chronic+wounds:+searching+for+an+optimal+animal+model+to+recapitulate+their+complexity.">Clinical challenges of chronic wounds: searching for an optimal animal model to recapitulate their complexity.</a> Dis Model Mech. 7, 1205-1213 (2014).</li><li>O'Brien, P. D., Sakowski, S. A., Feldman, E. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+models+of+diabetic+neuropathy.">Mouse models of diabetic neuropathy.</a> ILAR J. 54, 259-272 (2014).</li><li>Yamazaki, 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=Whole-Mount+Adult+Ear+Skin+Imaging+Reveals+Defective+Neuro-Vascular++Branching+Morphogenesis+in+Obese+and+Type+2+Diabetic+Mouse+Models.">Whole-Mount Adult Ear Skin Imaging Reveals Defective Neuro-Vascular Branching Morphogenesis in Obese and Type 2 Diabetic Mouse Models.</a> Sci Rep. 8, (2018).</li></ol>

本协议描述成人耳皮肤的全芒 immunonohistochemical 成像, 用于分析神经血管结构和免疫细胞分布。我们认为, 这种方法有许多实验优势, 研究人员的分支形态发生和模式的周围神经和血管, 以及三维分布的皮肤成分, 包括免疫细胞和头发毛囊.图像的结果可以用成像软件进行量化, 进一步定量分析。
正确地制备耳部皮肤对本议定书的成功至关重要。首先, euthanizing 后应仔细解剖耳朵皮...

皮肤由三层组成: 表皮、真皮和皮下组织。它已被作为一个模型系统来研究干细胞的维持, 分化和形态发生的发展, 以及再生, 肿瘤发生和炎症的成人。皮肤有丰富的血管化和支配, 使周围神经系统和血管系统的发育协调良好。
我们以前已经展示了一个完整的胚胎皮肤成像技术与多重标记研究完整的周围神经和血管, 包括其蜂窝组件1,2,3,4: 血管中的感觉轴突、交感神经轴突、神经的雪旺细胞、内皮细胞、毛细血管和血管平滑肌细胞 (VSMCs)。在血管生成过程中, 一个主要的毛细管网络经过密集的血管重塑, 发展成为一种分层的血管分支网络。在发育的真皮/皮下组织, 动脉分支与周围的感觉神经和静脉形成相邻的动脉。在分层的血管网络被 VSMCs 彻底覆盖后, 交感神经延伸沿和支配大口径血管1,5,6。尽管发展中的神经和血管系统之间的密切联系具有重要意义, 但一个主要问题是解决成人不同病理情况下神经血管网络的发生情况。三维高分辨率成像是必要的, 以了解发病机制, 以及解剖可识别的分支形态发生和模式。
组织切片染色法分析成年小鼠皮肤的神经细胞和血管形态。其他研究还使用了整个影像的皮肤来可视化周围神经和血管, 除了毛囊, 皮脂腺, 和 arrector 毛肌肉7,8,9。然而, 成人皮肤的厚度使得它很难分析整个深度的皮肤。
在本研究中, 我们开发了一种新的高分辨率的成人耳皮肤的全安装成像, 以克服这些挑战。耳朵皮肤是容易接近的解剖和后续整个安装成像的皮肤在其整个深度。因此, 它是一个简单的和高度重复的方法, 可用于比较三维结构的周围神经和血管系统的皮肤, 与综合量化测量。结果表明, 成人皮肤中保留了大口径血管对周围感觉和交感神经的定位。该协议的目标是可视化分支形态发生, 以及周围神经和血管的模式, 以及在各种情况下, 如炎症和成年小鼠模型的细胞水平上的免疫细胞分布。再生.

<table>
	<tbody>
		<tr>
			<td>10 x Phosphate Buffered Saline</td>
			<td>KD Medical</td>
			<td>RGE-3210</td>
			<td>PBS, without Ca2+/Mg2+</td>
		</tr>
		<tr>
			<td>Hank&#8217;s Balanced Salt Solution</td>
			<td>Gibco</td>
			<td>14025-092</td>
			<td>HBSS, with Ca2+/Mg2+</td>
		</tr>
		<tr>
			<td>16% Paraformaldehyde</td>
			<td>Electron Microscopy Sciences</td>
			<td>15710</td>
			<td>PFA, fixative, diluted in PBS</td>
		</tr>
		<tr>
			<td>Triton X-100</td>
			<td>Sigma</td>
			<td>X100</td>
			<td>Detergent</td>
		</tr>
		<tr>
			<td>Normal goat serum</td>
			<td>Gibco</td>
			<td>16210064</td>
			<td>Component of blocking/washing buffer</td>
		</tr>
		<tr>
			<td>Normal donkey serum</td>
			<td>Jackson Immuno research</td>
			<td>017-000-121</td>
			<td>Component of blocking/washing buffer</td>
		</tr>
		<tr>
			<td>Curved fine tweezers</td>
			<td>Dumont</td>
			<td>RS-5047</td>
			<td></td>
		</tr>
		<tr>
			<td>Curved tweezers</td>
			<td>Integra Miltex Vantage</td>
			<td>V918-782, V918-784</td>
			<td></td>
		</tr>
		<tr>
			<td>Filter Unit 0.45 mm</td>
			<td>Thermo Scientific</td>
			<td>157-0045</td>
			<td>For filtration</td>
		</tr>
		<tr>
			<td>1 mL syringe</td>
			<td>Coviden</td>
			<td>8881501400</td>
			<td>For filtration</td>
		</tr>
		<tr>
			<td>Syringe filter Unit 0.22 mm</td>
			<td>Millex-GV</td>
			<td>SLGVR04NL</td>
			<td>For filtration</td>
		</tr>
		<tr>
			<td>ProLong Gold</td>
			<td>Thermo Scientific</td>
			<td>P36934</td>
			<td>Anti-fade mounting medium</td>
		</tr>
		<tr>
			<td>Nail Polish</td>
			<td>Electron Microscopy Sciences</td>
			<td>72180</td>
			<td>For sealing</td>
		</tr>
		<tr>
			<td>Dissecting microscope</td>
			<td>Leica</td>
			<td>MZ95</td>
			<td></td>
		</tr>
		<tr>
			<td>Confocal microscope</td>
			<td>Leica</td>
			<td>TCS SP5</td>
			<td></td>
		</tr>
		<tr>
			<td>Photoshop CC 2017</td>
			<td>Adobe</td>
			<td></td>
			<td>Graphics editor software</td>
		</tr>
		<tr>
			<td>Illustrator CC 2017</td>
			<td>Adobe</td>
			<td></td>
			<td>Graphics editor software</td>
		</tr>
		<tr>
			<td>Image J</td>
			<td>NIH</td>
			<td></td>
			<td>Image processing software</td>
		</tr>
		<tr>
			<td>Anti-PECAM-1 (CD31) antibody</td>
			<td>Millipore</td>
			<td>MAB1398Z</td>
			<td>Hamster IgG, vascular endothelial cell marker, 1:300</td>
		</tr>
		<tr>
			<td>Anti-PECAM-1 (CD31) antibody</td>
			<td>BD Pharmingen</td>
			<td>553369</td>
			<td>Rat IgG2a kappa, vascular endothelial cell marker, 1:300</td>
		</tr>
		<tr>
			<td>Anti-aSMA antibody conjugated with cy-3</td>
			<td>Sigma</td>
			<td>C6198</td>
			<td>Mouse IgG2a, vascular smooth muscle cell marker, 1:500</td>
		</tr>
		<tr>
			<td>Anti-EphB1 antibody</td>
			<td>Santa Cruz</td>
			<td>sc-9319</td>
			<td>Goat polyclonal, venous endothelial cell marker, 1:100</td>
		</tr>
		<tr>
			<td>Anti-neuron-specific Class III b-tubulin (Tuj1)</td>
			<td>Abcam</td>
			<td>AB18207</td>
			<td>Tuj1, Rabbit polyclonal IgG, pan-axonal marker, 1:500</td>
		</tr>
		<tr>
			<td>Anti-Tuj1 antibody</td>
			<td>Covance</td>
			<td>MMS-435P</td>
			<td>Mouse IgG2a, pan-axonal marker, 1:500</td>
		</tr>
		<tr>
			<td>Anti-MBP antibody</td>
			<td>Abcam</td>
			<td>AB40390</td>
			<td>Rabbit polyclonal IgG, myelination marker, 1:200</td>
		</tr>
		<tr>
			<td>Anti-Tyrosine Hydroxylase antibody</td>
			<td>Chemicon</td>
			<td>AB152</td>
			<td>Rabbit polyclonal, sympathetic neuron marker, 1:500</td>
		</tr>
		<tr>
			<td>Anti-Peripherin antibody</td>
			<td>Chemicon</td>
			<td>AB1530</td>
			<td>Rabbit polyclonal, peripheral neuron marker, 1:1000</td>
		</tr>
		<tr>
			<td>Anti-CD11b antibody</td>
			<td>Bio-Rad</td>
			<td>MCA74G</td>
			<td>Rat IgG2b, inflammatory cell marker (macrophages), 1:50</td>
		</tr>
		<tr>
			<td>Anti-CD45 antibody</td>
			<td>Thermo Fisher Scientific</td>
			<td>14-0451-85</td>
			<td>Rat IgG2b kappa, pan-hematopoietic cell marker, 1:500</td>
		</tr>
		<tr>
			<td>Anti-CD3 antibody</td>
			<td>Bio-Rad</td>
			<td>MCA1477T</td>
			<td>Rat IgG1, immune cell marker, 1:100</td>
		</tr>
		<tr>
			<td>Anti-CD45R (B220) antibody</td>
			<td>Thermo Fisher Scientific</td>
			<td>14-0452</td>
			<td>Rat IgG2a kappa, inflammatory cell marker, 1:200</td>
		</tr>
		<tr>
			<td>Anti-GFP antibody</td>
			<td>Thermo Fisher Scientific</td>
			<td>A11122</td>
			<td>Rabbit polyclonal, 1:300</td>
		</tr>
		<tr>
			<td>Anti-GFP antibody</td>
			<td>Abcam</td>
			<td>Ab13970</td>
			<td>Chicken polyclonal, 1:500</td>
		</tr>
		<tr>
			<td>Anti-b-gal antibody</td>
			<td>Cappel</td>
			<td>55976</td>
			<td>Rabbit polyclonal, 1:5000</td>
		</tr>
		<tr>
			<td>Anti-RFP antibody</td>
			<td>Abcam</td>
			<td>Ab62341</td>
			<td>Rabbit polyclonal, 1:300</td>
		</tr>
		<tr>
			<td>Goat anti-rabbit IgG (H+L) Alexa 488</td>
			<td>Thermo Fisher Scientific</td>
			<td>A11034</td>
			<td>Rabbit polyclonal secondary antibody, 1:250</td>
		</tr>
		<tr>
			<td>Goat anti-hamster IgG (H+L) Alexa 647</td>
			<td>Jackson Immuno research</td>
			<td>127-605-160</td>
			<td>Hamster polyclonal secondary antibody, 1:250</td>
		</tr>
		<tr>
			<td>Goat anti-rat IgG (H+L) Alexa 594</td>
			<td>Jackson Immuno research</td>
			<td>112-585-167</td>
			<td>Rat polyclonal secondary antibody, 1:250</td>
		</tr>
		<tr>
			<td>Goat anti-mouse IgG2a Alexa 633</td>
			<td>Thermo Fisher Scientific</td>
			<td>A21136</td>
			<td>Mouse IgG2a secondary antibody, 1:250</td>
		</tr>
	</tbody>
</table>

whole-mount confocal microscopy for adult ear skin: a model system to study neuro-vascular branching morphogenesis and immune cell distribution

在这里, 我们提出了一个完整的成人耳皮肤成像技术的协议, 以研究全面的三维神经血管分支形态发生和模式, 以及免疫细胞分布的细胞水平。对成人组织周围神经和血管解剖结构的分析, 为了解创伤愈合等病理条件下的功能性神经血管接线和神经血管变性提供了一些见解。作为一个高度信息的模型系统, 我们把我们的研究集中在成人耳皮肤, 这是容易被解剖。我们的简单和可重现的协议提供了一个准确的描述整个皮肤的细胞成分, 如周围神经 (感觉轴突, 交感神经轴突, 和雪旺细胞), 血管 (内皮细胞和血管平滑肌细胞) 和炎症细胞。我们相信这项议定书将为调查不同病理条件下成人耳皮肤的形态学异常和周围神经和血管的炎症铺平道路。

成年小鼠后耳皮肤 (图 1A) 和前耳皮肤 (图 1B) 与αSMA (红色)、Tuj1 (绿色) 和 PECAM-1 (蓝色) 的抗体 immunostained。后皮肤 immunostained 研究神经免疫分布, 使用抗体 CD11b (红色) 和 MBP (绿色), 连同 Tuj1 (蓝色) (图 2A)。在单个细胞分辨率 (图 2B) 中检测到 CD11b+炎症细胞 (包括巨噬细胞) 的分?...

Watch this Scientific Journal Video about Whole-mount Confocal Microscopy for Adult Ear Skin: A Model System to Study Neuro-vascular Branching Morphogenesis and Immune Cell Distribution at JoVE.com

Whole-mount Confocal Microscopy for Adult Ear Skin: A Model System to Study Neuro-vascular Branching Morphogenesis and Immune Cell Distribution

在这里, 我们描述了一个高分辨率全安装成像方法在整个成年小鼠耳皮肤, 这使我们能够想象分支形态发生和模式的周围神经和血管, 以及免疫细胞分布。

成人耳皮肤全安装共焦显微术: 一种研究神经血管分支形态发生和免疫细胞分布的模型系统

Here, we present a protocol of a whole-mount adult ear skin imaging technique to study comprehensive three-dimensional neuro-vascular branching ...

Research

JoVE Journal

Developmental Biology

Live Imaging of Innate Immune and Preneoplastic Cell Interactions Using an Inducible Gal4/UAS Expression System in Larval Zebrafish Skin

先天免疫和癌前细胞间的相互作用使用可诱导的Gal4 / UAS表达系统在斑马鱼幼体皮肤实时成像

Here we describe a method to conditionally induce epithelial cell transformation by the use of the 4-Hydroxytamoxifen (4-OHT) inducible KalTA4-ERT2/UAS ...

科研

发育生物学

Epigenetic Conversion as a Safe and Simple Method to Obtain Insulin-secreting Cells from Adult Skin Fibroblasts

后生转换为安全和简单的方法来获得成人皮肤成纤维细胞胰岛素分泌细胞

Regenerative medicine requires new, fully functional cells that are delivered to patients in order to repair degenerated or damaged tissues. When such ...

Horizontal Whole Mount: A Novel Processing and Imaging Protocol for Thick, Three-dimensional Tissue Cross-sections of Skin

水平整体安装：厚三维组织横截面皮肤的新型加工和成像方案

Processing a tissue of interest to generate a microscopic image that supports a scientific argument can be challenging. The acquisition of high-quality ...

Quantitative Whole-mount Immunofluorescence Analysis of Cardiac Progenitor Populations in Mouse Embryos

小鼠胚胎心肌内祖群的定量全贴免疫荧光分析

The use of ever-advancing imaging techniques has contributed broadly to our increased understanding of embryonic development. Pre-implantation development ...

A Simplified and Efficient Method to Isolate Primary Human Keratinocytes from Adult Skin Tissue

一种简便有效的成人皮肤组织中人角质形成细胞的分离方法

Primary human keratinocytes isolated from fresh skin tissues and their expansion in vitro have been widely used for laboratory research and for clinical ...

Visualizing the Node and Notochordal Plate In Gastrulating Mouse Embryos Using Scanning Electron Microscopy and Whole Mount Immunofluorescence

用扫描电镜和全贴免疫荧光技术可视化小鼠胚胎中的节点和外型板

The post-implantation mouse embryo undergoes major shape changes after the initiation of gastrulation and morphogenesis. A hallmark of morphogenesis is ...

An In Vitro Model of a Parallel-Plate Perfusion System to Study Bacterial Adherence to Graft Tissues

An In Vitro Model of a Parallel-Plate Perfusion System to Study Bacterial Adherence to Graft Tissues

平行板灌注系统的体外模型研究了细菌对移植组织的粘附性

Various valved conduits and stent-mounted valves are used for right ventricular outflow tract (RVOT) valve replacement in patients with congenital heart ...

Analysis of Cardiac Chamber Development During Mouse Embryogenesis Using Whole Mount Epifluorescence

全山荧光在小鼠胚胎发生过程中心脏室发育的分析

The goal of this protocol is to describe a method for the dissection of mouse embryos and visualization of embryonic mouse ventricular chambers during ...

A Layered Mounting Method for Extended Time-Lapse Confocal Microscopy of Whole Zebrafish Embryos

全斑马鱼胚胎的延长时移共聚焦显微镜的分层安装方法

Dynamics of development can be followed by confocal time-lapse microscopy of live transgenic zebrafish embryos expressing fluorescence in specific tissues ...

Whole-Mount In Situ Hybridization in Zebrafish Embryos and Tube Formation Assay in iPSC-ECs to Study the Role of Endoglin in Vascular Development

斑马鱼胚胎全山原位杂交和管状测定在iPSC-ECs中研究恩多林在血管发育中的作用

Vascular development is determined by the sequential expression of specific genes, which can be studied by performing in situ hybridization assays in ...