Name: contrast imaging in mouse embryos using high-frequency ultrasound
Uploaded: 2015-03-04T14:00:00
Description: Watch this Scientific Journal Video about Contrast Imaging in Mouse Embryos Using High-frequency Ultrasound at JoVE.com

This work was supported by the Terry Fox Program of the National Cancer Institute of Canada.

注：この研究で行った実験手順はサニーブルック研究所（トロント、オンタリオ州、カナダ）での動物実験委員会によって承認された。動物の人道的な扱いのための手順は、すべての回で観察されなければならない。これは、治験責任医師は、超音波撮像システムの基本動作の訓練を受けているものとする。このプロトコルは、2人で最適に動作。 1.動物モデル<ol><li>メ...

<ol>
	<li>Voigt, J. U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ultrasound+molecular+imaging.">Ultrasound molecular imaging.</a> Methods. 48 (2), 92-97 (2009).</li><li>Klibanov, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preparation+of+targeted+microbubbles:+Ultrasound+contrast+agents+for+molecular+imaging.">Preparation of targeted microbubbles: Ultrasound contrast agents for molecular imaging.</a> Medical Biological Engineering Computing. 47 (8), 875-882 (2009).</li><li>Cosgrove, D., Lassau, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Imaging+of+perfusion+using+ultrasound.">Imaging of perfusion using ultrasound.</a> European Journal Of Nuclear Medicine And Molecular Imaging. 37 (S1), 65 (2010).</li><li>Williams, 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=Dynamic+microbubble+contrast-enhanced+US+to+measure+tumor+response+to+targeted+therapy:+A+proposed+clinical+protocol+with+results+from+renal+cell+carcinoma+patients+receiving+antiangiogenic+therapy.">Dynamic microbubble contrast-enhanced US to measure tumor response to targeted therapy: A proposed clinical protocol with results from renal cell carcinoma patients receiving antiangiogenic therapy.</a> Radiology. 260 (2), 581 (2011).</li><li>Burns, P. N., Wilson, S. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Focal+liver+masses:+Enhancement+patterns+on+contrast-enhanced+Images+-+Concordance+of+US+scans+with+CT+scans+and+MR+images.">Focal liver masses: Enhancement patterns on contrast-enhanced Images - Concordance of US scans with CT scans and MR images.</a> Radiology. 242 (1), 162 (2006).</li><li>Phoon, C. K. L., Aristizabal, O., Turnbull, D. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=40+MHz+doppler+characterization+of+umbilical+and+dorsal+aortic+Blood+flow+in+the+early+mouse+embryo.+Ultrasound.">40 MHz doppler characterization of umbilical and dorsal aortic Blood flow in the early mouse embryo. Ultrasound.</a> In Medicine And Biology. 26 (8), 1275-1283 (2000).</li><li>Phoon, C. K. L., Aristizabal, O., Turnbull, D. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spatial+velocity+profile+in+mouse+embryonic+aorta+and+doppler-derived+volumetric+flow:+A+preliminary+model.">Spatial velocity profile in mouse embryonic aorta and doppler-derived volumetric flow: A preliminary model.</a> Am J Physiol Heart Circ Physiol. 283, H908-H916 (2002).</li><li>Aristizábal, O., Williamson, R., Turnbull, D. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> 12A-4 in vivo 3D contrast-enhanced imaging of the embryonic mouse vasculature. Paper presented at Ultrasonics Symposium. , (2007).</li><li>Bartelle, B. B., 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+genetic+approach+for+in+vivo+vascular+imaging+in+mice.">Novel genetic approach for in vivo vascular imaging in mice.</a> Circ.Res. 110 (7), 938-947 (2012).</li><li>Endoh, 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=Fetal+gene+transfer+by+intrauterine+injection+with+microbubble-enhanced+ultrasound.">Fetal gene transfer by intrauterine injection with microbubble-enhanced ultrasound.</a> Molecular Therapy. 5 (5), 501-508 (2002).</li><li>Yamada, M., Hatta, T., Otani, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+exo+utero+development+system:+Protocol+and+troubleshooting.">Mouse exo utero development system: Protocol and troubleshooting.</a> Congenital Anomalies. 48 (4), 183-187 (2008).</li><li>Denbeigh, J. M., Nixon, B. A., Hudson, J. M., Purin, M. C., Foster, F. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=VEGFR2-targeted+molecular+imaging+in+the+mouse+embryo:+An+alternative+to+the+tumor+model.">VEGFR2-targeted molecular imaging in the mouse embryo: An alternative to the tumor model.</a> Ultrasound in medicine and biology. 40 (2), 389-399 (2014).</li><li>Paauwe, M., Dijke, t. e. n., P, L. J. A. C., Hawinkels, <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endoglin+for+tumor+imaging+and+targeted+cancer+therapy.">Endoglin for tumor imaging and targeted cancer therapy.</a> Expert Opinion On Therapeutic Targets. 17 (4), 421-435 (2013).</li><li>Bourdeau, A., Faughnan, M. E., Letarte, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endoglin-deficient+mice,+a+unique+model+to+study+hereditary+hemorrhagic+telangiectasia.">Endoglin-deficient mice, a unique model to study hereditary hemorrhagic telangiectasia.</a> Trends Cardiovasc. Med. 10 (7), 279-285 (2000).</li><li>Whiteley, K. J., Adamson, S. L., Pfarrer, 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=Vascular+corrosion+casting+of+the+uteroplacental+and+fetoplacental+vasculature+in+mice.">Vascular corrosion casting of the uteroplacental and fetoplacental vasculature in mice.</a> Placenta And Trophoblast: Methods And Protocols. 121 (121), 371-392 (2006).</li><li>Kulandavelu, 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=Embryonic+and+neonatal+phenotyping+of+genetically+engineered+mice.">Embryonic and neonatal phenotyping of genetically engineered mice.</a> ILAR Journal. 47 (2), 103-10 (2006).</li><li>Kalaskar, V. K., Lauderdale, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+embryonic+development+in+a+serum-free+whole+embryo+culture+system.">Mouse embryonic development in a serum-free whole embryo culture system.</a> Journal of Visualized Experiments. 85, (2014).</li><li>Willmann, J. 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=Targeted+contrast-enhanced+ultrasound+imaging+of+tumor+angiogenesis+with+contrast+microbubbles+conjugated+to+integrin-binding+knottin+peptides.">Targeted contrast-enhanced ultrasound imaging of tumor angiogenesis with contrast microbubbles conjugated to integrin-binding knottin peptides.</a> The Journal of Nuclear Medicine. 51 (3), 433-440 (2010).</li><li>Deshpande, N., Ren, Y., Foygel, K., Rosenberg, J., Willmann, J. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tumor+angiogenic+marker+expression+levels+during+tumor+growth:+Longitudinal+assessment+with+molecularly+targeted+microbubbles+and+US+imaging.">Tumor angiogenic marker expression levels during tumor growth: Longitudinal assessment with molecularly targeted microbubbles and US imaging.</a> Radiology. 258 (3), 804-811 (2011).</li><li>Lyshchik, 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=Molecular+imaging+of+vascular+endothelial+growth+factor+receptor+2+expression+using+targeted+contrast-enhanced+high-frequency+ultrasonography.">Molecular imaging of vascular endothelial growth factor receptor 2 expression using targeted contrast-enhanced high-frequency ultrasonography.</a> Journal Of Ultrasound In Medicine. 26 (11), 1575-1586 (2007).</li><li>Jerkic, 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=Endoglin+regulates+nitric+oxide-dependent+vasodilatation.">Endoglin regulates nitric oxide-dependent vasodilatation.</a> The FASEB Journal. 18 (3), 609-611 (2004).</li><li>Denbeigh, J. M., Nixon, B. A., Lee, J. J. 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=.">.</a> Contrast-Enhanced Molecular Ultrasound Differentiates Endoglin Genotypes in Mouse Embryos. , (2014).</li><li>Adamson, S. L., Lu, Y., Whiteley, K. J., 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=Interactions+between+trophoblast+cells+and+the+maternal+and+fetal+circulation+in+the+mouse+placenta.">Interactions between trophoblast cells and the maternal and fetal circulation in the mouse placenta.</a> Dev Biol. 250, 358-35 (2002).</li><li>Needles, 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=Nonlinear+contrast+imaging+with+an+array-based+micro-ultrasound+system.+Ultrasound.">Nonlinear contrast imaging with an array-based micro-ultrasound system. Ultrasound.</a> Medicine Biology. 36 (12), 2097 (2010).</li><li>Watson, E. D., Cross, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+structures+and+transport+functions+in+the+mouse+placenta.">Development of structures and transport functions in the mouse placenta.</a> Physiology. 20 (3), 180-193 (2005).</li><li>Shalaby, F., Rossant, J., Yamaguchi, T. 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=Failure+of+blood-island+formation+and+vasculogenesis+in+Flk-1-deficient+mice.">Failure of blood-island formation and vasculogenesis in Flk-1-deficient mice.</a> Nature. 376, 62-66 (1995).</li><li>Kwee, L., Baldwin, H. S., Shen, H. 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=Defective+development+of+the+embryonic+and+extraembryonic+circulatory+systems+in+vascular+cell+adhesion+molecule+(VCAM-1)+deficient+mice.">Defective development of the embryonic and extraembryonic circulatory systems in vascular cell adhesion molecule (VCAM-1) deficient mice.</a> Development. 121, (1995).</li><li>Mercurio, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lessons+from+the+&#945;2+integrin+knockout+mouse.">Lessons from the &#945;2 integrin knockout mouse.</a> The American journal of pathology. , 161-163 (2002).</li><li>Hodivala-Dilke, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=&#945;v&#946;3+integrin+and+angiogenesis:+a+moody+integrin+in+a+changing+environment.">&#945;v&#946;3 integrin and angiogenesis: a moody integrin in a changing environment.</a> Curr Opin Cell Biol. 20, 514-519 (2008).</li><li>Pysz, M. A., Gambhir, S. S., Willmann, J. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+imaging:+current+status+and+emerging+strategies.">Molecular imaging: current status and emerging strategies.</a> Clinical radiology. 65, 500-516 (2010).</li><li>Cybulsky, M. I., Iiyama, K., Li, H., 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=A+major+role+for+VCAM-1,+but+not+ICAM-1,+in+early+atherosclerosis.">A major role for VCAM-1, but not ICAM-1, in early atherosclerosis.</a> J Clin Invest. 107, 1255-1262 (2001).</li><li>Xu, H., Gonzalo, J. A., St Pierre, ., 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=Leukocytosis+and+resistance+to+septic+shock+in+intercellular+adhesion+molecule+1-deficient+mice.">Leukocytosis and resistance to septic shock in intercellular adhesion molecule 1-deficient mice.</a> J Exp Med. 180, 95-109 (1994).</li><li>Gerwin, N., Gonzalo, J. A., Lloyd, 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=Prolonged+eosinophil+accumulation+in+allergic+lung+interstitium+of+ICAM-2-deficient+mice+results+in+extended+hyperresponsiveness.">Prolonged eosinophil accumulation in allergic lung interstitium of ICAM-2-deficient mice results in extended hyperresponsiveness.</a> Immunity. 10, 9-19 (1999).</li><li>Johnson, R. C., Mayadas, T. N., Frenette, P. 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=Blood+cell+dynamics+in+P-selectin-deficient+mice.">Blood cell dynamics in P-selectin-deficient mice.</a> Blood. 86, 1106-1114 (1995).</li><li>Corrigan, N., Brazil, D., McAuliffe, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-frequency+ultrasound+assessment+of+the+murine+heart+from+embryo+through+to+juvenile.">High-frequency ultrasound assessment of the murine heart from embryo through to juvenile.</a> Reproductive Sciences. 17 (2), 147-14 (2010).</li><li>Turnbull, D. H., Bloomfield, T. S., Baldwin, H. S., Foster, F. S., Joyner, A. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ultrasound+backscatter+microscope+analysis+of+early+mouse+embryonic+brain+development.">Ultrasound backscatter microscope analysis of early mouse embryonic brain development.</a> Proc Natl Acad Sci U S A. 92, 2239-2243 (1995).</li><li>Greco, A., Mancini, M. L., Gargiulo, 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=Ultrasound+Biomicroscopy+in+Small+Animal+Research:+Applications+in+Molecular+and+Preclinical+Imaging.">Ultrasound Biomicroscopy in Small Animal Research: Applications in Molecular and Preclinical Imaging.</a> Journal of Biomedicine and Biotechnology. 2012, (2012).</li><li>Pysz, M. A., Guracar, I., Foygel, K., Tian, L., Willmann, J. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+assessment+of+tumor+angiogenesis+using+real-time+motion-compensated+contrast-enhanced+ultrasound+imaging.">Quantitative assessment of tumor angiogenesis using real-time motion-compensated contrast-enhanced ultrasound imaging.</a> Angiogenesis. 15, 433-442 (2012).</li><li>Larina, I. V., 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+imaging+of+blood+flow+in+mammalian+embryos+using+doppler+swept-source+optical+coherence+tomography.">Live imaging of blood flow in mammalian embryos using doppler swept-source optical coherence tomography.</a> J.Biomed.Opt. 13 (6), 060506-06 (2008).</li><li>Garcia, M. D., Udan, R. S., Hadjantonakis, A. K., Dickinson, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Live imaging of mouse embryos. 4 (4), 104-10 (2011).</li><li>Teichert, 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=Endothelial+nitric+oxide+synthase+gene+expression+during+murine+embryogenesis.+Commencement+of+expression+in+the+embryo+occurs+with+the+establishment+of+a+unidirectional+circulatory+system.">Endothelial nitric oxide synthase gene expression during murine embryogenesis. Commencement of expression in the embryo occurs with the establishment of a unidirectional circulatory system.</a> Circulation Research. 103 (1), 24-33 (2008).</li><li>Walls, J. R., Coultas, L., Rossant, J., Henkelman, R. M. <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+analysis+of+vascular+development+in+the+mouse+embryo.">Three-dimensional analysis of vascular development in the mouse embryo.</a> PLoS One. 3 (8), (2008).</li></ol>

超音波造影剤は、後期妊娠マウス胚および非線形コントラスト画像に注射し、灌流パラメータを測定するために取得し、結合マイクロバブルを標的とした。胚の血管系内のマイクロバブルの成功イメージングは​​、多くの要因、最初のもの胚の生存率に依存していた。すべての機器および装置は、注入の開始に子宮から胚を単離するために必要な時間を最小にするために、事前に調製した?...

造影超音波イメージングは​​、血管環境を視覚化し、特徴付けるために、マイクロバブル造影剤を利用する。これらの物質は、微小循環、血管分布および心血管機能の非侵襲的評価を可能に。また、気泡表面の改質が可能な血管イベントの分子の超音波イメージングを行う血管形成、アテローム性動脈硬化症および炎症1,2の前臨床用途で実証されるように、内皮細胞のバイオマーカーに結合する標的化マイクロバブルをもたらすことができる。造影超音波は、したがって、健康と病気の血管の状態3-5に影響を与える複雑かつ多様な環境を識別するために使用することができる。 年の過去数には、マイクロバブルイメージングの有用性への関心は汎用性の高いマウス胚モデルに拡張しました。哺乳類の発生のモデルとして、胚の血管系へのマイクロバブルの導入は、生理的な強化開発循環系（ 例えば 、血流量、心拍出量）およびトランスジェニック例での研究と心疾患6,7の変異マウスモデルをターゲットとは、遺伝的要因が心血管機能を変化させる方法への洞察をもたらすことができる。実際には、胚の脳血管系の定量的および定性的な二次元分析は、既に8を達成している。さらに、マウス胚は、生体内での血管マーカーに標的マイクロバブルの結合を調べるための優れたモデルとして提示します。 Bartelle ら 9は 、例えば、Biotag-BirAをトランスジェニック胚に結合し、血管構造を検討対象と評価するために胚心臓の心室にアビジンマイクロバブルを導入しています。診療所にこの技術を翻訳において重要なベンチマーク - ヘテロ接合およびホモ接合性マウスモデルの生成は、分子の超音波の定量的性質を規定することを目的と腫瘍モデル研究のための代用として使用することができる。 8-10を介して露出し、単一の胚に心臓内注射を経由して胎児の血液循環に導入される。 子宮内注射では 、しかし、多くの課題に直面しています。これらは、注射の指導、母親と体外胚の動きに対抗する必要性、母親における血行動態の生存率の維持および11の出血による麻酔の合併症の長期的影響に対処体外胚を、含む。従って、調査の目的は、単離されたリビング後期胚12にマイクロバブルを注入するための技術を開発することであった。このオプションは、噴射制御やポジショニング、障害物のない撮像面の再現性、および単純化された画像解析と定量化の面でより多くの自由を提供しています。 本研究では、foの生きているマウスの胚にマイクロバブルを注入するための新規な手順を概説Rマイクロバブル速度論的挙動を研究するとの目的は、内因性の内皮表面マーカーに結合するマイクロバブルをターゲットに勉強。非線形コントラスト特有の超音波イメージングは​​、ピークエンハンスメント（PE）、ウォッシュアウト速度および時間孤立E17.5胚において（TTP）をピークに含む基本的な灌流パラメータの数を測定するために使用される。我々はまた、活性な血管新生13の部位で血管内皮細胞におけるその高い発現をエンドグリンは、臨床的に関連する標的で機能するトランスジェニックマウスモデルの胚グリン損失分子音波の定量的性質を評価するための胚のモデルの妥当性を実証する。エンドグリンを標的（MB E）の付着、ラットアイソタイプIgGを2コントロール（MB C）および非標的（MB U）マイクロバブルは、ヘテロ接合グリン（ 工学 +/-）で評価され、ホモ接合エンドグリン（ 工 + / +）の胚を表す。目標とビンディの分析ngは、分子の超音波は、エンドグリン遺伝子型を区別し、定量化可能な分子の超音波レベルに受容体密度に関連することが可能であることを明らかにする。

<table border="0" cellpadding="0" cellspacing="0" width="1477">
	<tbody>
		<tr height="20">
			<td height="20" style="height:20px;width:419px;">Reagents</td>
			<td style="width:215px;">Company</td>
			<td style="width:207px;">Catalog Number</td>
			<td style="width:639px;">Comments/Description</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:419px;"></td>
			<td style="width:215px;"></td>
			<td style="width:207px;"></td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Antibodies (biotinylated, eBioscience)</td>
			<td style="width:215px;"></td>
			<td style="width:207px;"></td>
			<td>Antibody choice depends on the experiment</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;">&#160;&#160;&#160;&#160;&#160; rat isotype IgG2 control</td>
			<td style="width:215px;">eBioscience</td>
			<td style="width:207px;">13-4321-85</td>
			<td>This antibody/microbubble combination is often required as experimental control&#160;</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">&#160;&#160;&#160;&#160;&#160; biotin anti-mouse CD309</td>
			<td style="width:215px;">eBioscience</td>
			<td style="width:207px;">13-5821-85</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Biotinylated rat MJ 7/18 antibody to mouse endoglin</td>
			<td style="width:215px;">In house hybridoma</td>
			<td style="width:207px;"></td>
			<td>Outside antibodies may also be appropriate: we&#160; have used eBioscience (13-1051-85 ) in the past</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Distilled water</td>
			<td style="width:215px;"></td>
			<td style="width:207px;"></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Embryo media</td>
			<td style="width:215px;"></td>
			<td style="width:207px;"></td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">&#160;&#160;&#160;&#160; 500 mL Dulbecco&#8217;s Modified Eagle&#8217;s Medium with high glucose</td>
			<td>Sigma</td>
			<td>D5796</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">&#160;&#160;&#160;&#160; 50 mL Fetal Bovine Serum</td>
			<td>ATCC</td>
			<td style="width:207px;">30-2020</td>
			<td>lot # 7592456</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">&#160;&#160;&#160;&#160; Hepes&#160;</td>
			<td style="width:215px;">Gibco</td>
			<td style="width:207px;">15630</td>
			<td>5mL, 1M</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">&#160;&#160;&#160;&#160; Penicillin-Streptomycin&#160;</td>
			<td style="width:215px;">Gibco</td>
			<td style="width:207px;">15140-122</td>
			<td>5 mL, 10,000 units Pen., 10,000 ug Strep</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Ethanol, 70%</td>
			<td style="width:215px;"></td>
			<td style="width:207px;"></td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Ice</td>
			<td style="width:215px;"></td>
			<td style="width:207px;"></td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Paraformaldehyde</td>
			<td style="width:215px;">Sigma</td>
			<td style="width:207px;">76240</td>
			<td>4%</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Phosphate Buffered Saline [1x]&#160;</td>
			<td style="width:215px;">Sigma</td>
			<td style="width:207px;">D8537</td>
			<td>1x, w/o calcium chloride &#38; magnesium chloride</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Pregnant mouse, CD-1</td>
			<td style="width:215px;">Charles River Laboratories Inc.&#160;</td>
			<td style="width:207px;"></td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">0.9% sodium chloride (saline)</td>
			<td style="width:215px;">Hospira</td>
			<td style="width:207px;">0409-7984-11</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Ultrasound contrast agent, target ready and untargeted</td>
			<td style="width:215px;">MicroMarker; VisualSonics Inc.</td>
			<td style="width:207px;"></td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Ultrasound gel (Aquasonic 100, colourless)</td>
			<td style="width:215px;">CSP Medical</td>
			<td style="width:207px;">133-1009</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;"></td>
			<td style="width:215px;"></td>
			<td style="width:207px;"></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Equipment</td>
			<td style="width:215px;"></td>
			<td style="width:207px;"></td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;"></td>
			<td style="width:215px;"></td>
			<td style="width:207px;"></td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Cell culture plates (4) :&#160; 100x20 mm</td>
			<td style="width:215px;">Fisher Scientific</td>
			<td style="width:207px;">08-772-22</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Cell culture plates (12) : 60x15 mm</td>
			<td style="width:215px;">Sigma</td>
			<td style="width:207px;">D8054</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:419px;">Centrifuge</td>
			<td>Sorvall Legend RT centrifuge&#160;</td>
			<td style="width:207px;"></td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Conical tubes, 50 mL BD Falcon</td>
			<td style="width:215px;">VWR</td>
			<td style="width:207px;">21008-938</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Diluent</td>
			<td style="width:215px;">Beckman Coulter</td>
			<td style="width:207px;">Isoton II Diluent, 8448011</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Dissection scissors (Wagner)</td>
			<td style="width:215px;">Fine Science Tools</td>
			<td style="width:207px;">Wagner 14068-12</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Forceps (2), Dumont SS (0.10x0.06 mm)</td>
			<td style="width:215px;">Fine Science Tools</td>
			<td style="width:207px;">11200-33</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Forceps, splinter</td>
			<td style="width:215px;">VWR</td>
			<td style="width:207px;">25601-134</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Glass beaker, 2 L (Griffin Beaker)</td>
			<td style="width:215px;">VWR</td>
			<td style="width:207px;">89000-216</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Glass capillaries, 1x90 mm GD-1 with filament</td>
			<td style="width:215px;">Narishige</td>
			<td style="width:207px;">GD-1</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Glass needle puller</td>
			<td style="width:215px;">Narishige</td>
			<td style="width:207px;">PN-30</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Gloves</td>
			<td style="width:215px;">Ansell</td>
			<td style="width:207px;">4002</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Gross anatomy probe</td>
			<td style="width:215px;">Fine Science Tools</td>
			<td style="width:207px;">10088-15</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Hot plate</td>
			<td style="width:215px;">VWR</td>
			<td style="width:207px;">89090-994</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Ice bucket</td>
			<td style="width:215px;">Cole Parmer</td>
			<td style="width:207px;">RK 06274-01</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Imaging Platform</td>
			<td style="width:215px;">VisualSonics Inc.</td>
			<td style="width:207px;">Integrated Rail System</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Light source, fiber-optic</td>
			<td style="width:215px;">Fisher Scientific</td>
			<td style="width:207px;">12-562-36</td>
			<td>Ideally has adjustable arms</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Luers (12), polypropylene barbed female &#188;-28 UNF thread</td>
			<td style="width:215px;">Cole Parmer</td>
			<td style="width:207px;">45500-30</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Micro-ultrasound system, high-frequency</td>
			<td style="width:215px;">VisualSonics Inc.</td>
			<td style="width:207px;">Vevo2100</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Needles, 21 gauge&#160; (1&#8221;)</td>
			<td style="width:215px;">VWR</td>
			<td style="width:207px;">305165</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Particle size analyzer</td>
			<td style="width:215px;">Beckman Coulter</td>
			<td style="width:207px;">Multisizer 3 Coulter Counter</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Perforated spoon (Moria)</td>
			<td style="width:215px;">Fine Science Tools</td>
			<td style="width:207px;">MC 17 10373-17</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Pins (6), black anodized minutien 0.15 mm</td>
			<td style="width:215px;">Fine Science Tools</td>
			<td style="width:207px;">26002-15</td>
			<td></td>
		</tr>
		<tr height="57">
			<td height="57" style="height:57px;">Pipettors [2-20 uL, 20-200uL, 100-1000uL]</td>
			<td style="width:215px;">Eppendorf</td>
			<td style="width:207px;">Research Plus&#160; adjustable 3120000038;&#160;&#160;&#160;&#160;&#160;&#160; 3120000054;&#160;&#160;&#160;&#160;&#160;&#160; 3120000062</td>
			<td style="width:639px;"></td>
		</tr>
		<tr height="38">
			<td height="38" style="height:38px;">Pipettor tips [2-200uL, 50-1000uL]</td>
			<td style="width:215px;">Eppendorf</td>
			<td style="width:207px;">epT.I.P.S.&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; 22491334;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; 022491351</td>
			<td style="width:639px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Scissors</td>
			<td style="width:215px;"></td>
			<td style="width:207px;"></td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Sylgard 184 Silicone Elastomer Kit</td>
			<td style="width:215px;">Dow Corning</td>
			<td style="width:207px;"></td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Tubing, Tygon laboratory 1/32x3/32&#8221;</td>
			<td style="width:215px;">VWR</td>
			<td style="width:207px;">63010-007</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Wooden applicator stick (swab, cotton head)</td>
			<td style="width:215px;">VWR</td>
			<td style="width:207px;">CA89031-270</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Surgical microscope 5-8x magnification</td>
			<td style="width:215px;">Fisher Scientific</td>
			<td style="width:207px;">Steromaster</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Syringes, 1 mL Normject</td>
			<td style="width:215px;">Fisher</td>
			<td style="width:207px;">14-817-25</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Syringes (10), 30 mL</td>
			<td style="width:215px;">VWR</td>
			<td style="width:207px;">CA64000-041</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Syringe infusion pump&#160;</td>
			<td style="width:215px;">Bio-lynx&#160;</td>
			<td style="width:207px;">NE-1000</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;">Thermometer, -20-110oC</td>
			<td style="width:215px;">VWR</td>
			<td style="width:207px;">89095-598</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Timer</td>
			<td style="width:215px;">VWR</td>
			<td style="width:207px;">33501-418</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Tubes, Eppendorf</td>
			<td style="width:215px;">VWR</td>
			<td style="width:207px;">20170-577</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Tube racks (3)</td>
			<td style="width:215px;">VWR</td>
			<td style="width:207px;">82024-462</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Ultrasound transducer, 20 MHz</td>
			<td style="width:215px;">VisualSonics Inc.</td>
			<td style="width:207px;">MS250</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Vannas-Tubingen, angled up</td>
			<td style="width:215px;">Fine Science Tools</td>
			<td style="width:207px;">15005-08</td>
			<td></td>
		</tr>
	</tbody>
</table>

contrast imaging in mouse embryos using high-frequency ultrasound

Ultrasound contrast-enhanced imaging can convey essential quantitative information regarding tissue vascularity and perfusion and, in targeted applications, facilitate the detection and measure of vascular biomarkers at the molecular level. Within the mouse embryo, this noninvasive technique may be used to uncover basic mechanisms underlying vascular development in the early mouse circulatory system and in genetic models of cardiovascular disease. The mouse embryo also presents as an excellent model for studying the adhesion of microbubbles to angiogenic targets (including vascular endothelial growth factor receptor 2 (VEGFR2) or &#945;v&#946;3) and for assessing the quantitative nature of molecular ultrasound. We therefore developed a method to introduce ultrasound contrast agents into the vasculature of living, isolated embryos. This allows freedom in terms of injection control and positioning, reproducibility of the imaging plane without obstruction and motion, and simplified image analysis and quantification. Late gestational stage (embryonic day (E)16.6 and E17.5) murine embryos were isolated from the uterus, gently exteriorized from the yolk sac and microbubble contrast agents were injected into veins accessible on the chorionic surface of the placental disc. Nonlinear contrast ultrasound imaging was then employed to collect a number of basic perfusion parameters (peak enhancement, wash-in rate and time to peak) and quantify targeted microbubble binding in an endoglin mouse model. We show the successful circulation of microbubbles within living embryos and the utility of this approach in characterizing embryonic vasculature and microbubble behavior.

子宮外でのマウス胚への超音波造影剤の注入は、注入および関連する超音波撮像にわたって生存能力の子宮およびメンテナンスから後期妊娠期胚生活の正常な分離に依存する。胚体外に位置決めされた後、図1に示すように、胚の血管系への造影剤の慎重な注入が可能である。 E17.5マウス胚の典型的なBモード超音波画像は、 図2Aに示されている。マイクロバブ...

Watch this Scientific Journal Video about Contrast Imaging in Mouse Embryos Using High-frequency Ultrasound at JoVE.com

Contrast Imaging in Mouse Embryos Using High-frequency Ultrasound

ここでは、生きている、孤立した後期妊娠期のマウス胚に超音波マイクロバブル造影剤を注入するためのプロトコルを提示する。この方法は、灌流パラメータ及びコントラスト強調高周波超音波イメージングを用いて、胚内の血管の分子マーカーの研究を可能にする。

高周波超音波を用いて、マウス胚におけるコントラストイメージング

Ultrasound contrast-enhanced imaging can convey essential quantitative information regarding tissue vascularity and perfusion and, in targeted ...

Research

JoVE Journal

Developmental Biology

Analyzing In Vivo Cell Migration using Cell Transplantations and Time-lapse Imaging in Zebrafish Embryos

Analyzing In Vivo Cell Migration using Cell Transplantations and Time-lapse Imaging in Zebrafish Embryos

分析<em&gt;インビボ</emゼブラフィッシュ胚における細胞移植とタイムラプスイメージングを使用して&gt;細胞移動

Cell migration is key to many physiological and pathological conditions, including cancer metastasis. The cellular and molecular bases of cell migration ...

Live Imaging of Mouse Secondary Palate Fusion

マウス二次口蓋融合のライブイメージング

The fusion of the secondary palatal shelves to form the intact secondary palate is a key process in mammalian development and its disruption can lead to ...

A Novel Use of Three-dimensional High-frequency Ultrasonography for Early Pregnancy Characterization in the Mouse

マウスの早い妊娠の特性の三次元の高周波超音波の新規な使用

High-frequency ultrasonography (HFUS) is a common method to non-invasively monitor the real-time development of the human fetus in utero. The mouse is ...

Fetal Mouse Cardiovascular Imaging Using a High-frequency Ultrasound (30/45MHZ) System

Fetal Mouse Cardiovascular Imaging Using a High-frequency Ultrasound 30/45MHZ System

心血管イメージング (30/45 MHZ) の高周波超音波を用いた胎児マウス

Congenital heart defects (CHDs) are the most common cause of childhood morbidity and early mortality. Prenatal detection&#160;of the underlying molecular ...

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

ノードと走査型電子顕微鏡と蛍光抗体法マウント全体を使用して Gastrulating のマウス胚における脊索板を可視化

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

High Frequency Ultrasound for the Analysis of Fetal and Placental Development In Vivo

High Frequency Ultrasound for the Analysis of Fetal and Placental Development In Vivo

体内の胎児と胎盤の開発の分析のための高周波超音波

Ultrasound imaging is a widespread method used to detect organ anomalies and tumors in human and animal tissues. The method is non-invasive, harmless, and ...

Analysis of Congenital Heart Defects in Mouse Embryos Using Qualitative and Quantitative Histological Methods

定性・定量的組織学的手法を用いたマウス胚における先天性心不全の解析

Congenital heart defects (CHD) are the most common type of birth defect in humans, affecting up to 1% of all live births. However, the underlying causes ...

High-Frequency Ultrasound Echocardiography to Assess Zebrafish Cardiac Function

ゼブラフィッシュ心臓機能を評価する高周波超音波心エコー検査

The zebrafish (Danio rerio) has become a very popular model organism in cardiovascular research, including human cardiac diseases, largely due to its ...

Imaging Intranuclear Actin Rods in Live Heat Stressed Drosophila Embryos

Imaging Intranuclear Actin Rods in Live Heat Stressed Drosophila Embryos

ライブ熱ストレスショ ウジョウバエ 胚における核内アクチンロッドのイメージング

The purpose of this protocol is to visualize intranuclear actin rods that assemble in live Drosophila melanogaster embryos following heat stress. Actin ...

Is My Mouse Pregnant? High-Frequency Ultrasound Assessment

私のマウスは妊娠していますか?高周波超音波評価

The mouse is the mammalian animal model of choice for many human diseases and biological processes. Developmental biology often requires staged-pregnant ...