Name: retrograde perfusion and filling of mouse coronary vasculature as preparation for micro computed tomography imaging
Uploaded: 2012-02-10T13:00:00
Description: Watch this Scientific Journal Video about Retrograde Perfusion and Filling of Mouse Coronary Vasculature as Preparation for Micro Computed Tomography Imaging at JoVE.com

Biz protokol, Dr Michael Simons, Dr Kip Hauch ve genel tartışma için laboratuvarlarında her iki üyelerinin ilk denemeleri için Dr Kelly Stevens teşekkür ederim. Bu çalışma NIH hibe HL087513 ve P01 HL094374 tarafından destek.

1. Başlamadan önce hazırlıklar <ol><li> , Sırasıyla, PBS içerisinde Vazodilatör tampon (4 mg / L Papaverin + 1 gr / L PBS içinde Adenosin) veya 4% paraformaldehit (PFA) ile basınçlı perfüzyon aparatı her bir tarafında doldurun. </li><li> 1:100 Heparin 0.1 ml (5000U/ml hisse senedi) ile doldurma ve yukarı eğim ile ~ 120 derecelik bir açıyla iğne bükerek bir 1/2cc İnsülin Şırınga (bir srekli 29G ½ &quot;iğne ile) hazırlayın. Mı 0.3 ml doymuş KCl çözeltisi ile doldurulmuş 1 ml şırıng...

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Kardiyak doku çok yüksek metabolik talep vardır ve bu nedenle, koroner damar tarafından sağlanan kandan besin ve oksijen sabit bir besleme gerektirir. Damar darlığı ve tıkanma nedeniyle koroner fonksiyonu azaltmak koroner damarlarında, hastalıkları, doku hipoksisi ve iskemiye yol, myokard infarktüsü ve kalp kası onarılamaz hasar riski etkilenen hastalar koyabilirsiniz. Bu gemilerin hastalıklı devlet daha iyi anlaşılması gereklidir ve koroner damarların çalışma yeteneğimizi kritik damarların g?...

<html><table border="1"><tbody><tr><td> Reaktif Adı </td><td> Şirket </td><td> Katalog numarası </td><td> Yorumlar </td></tr><tr><td> 1 ml şırınga </td><td> <a href="http://www.bd.com/" target="_blank">Becton Dickinson</a> </td><td> BD-309.602 </td><td></td></tr><tr><td> Srekli 29G ½ &#39;iğne ile 1/2cc insülin şırıngaları </td><td> <a href="http://www.bd.com/" target="_blank">Becton Dickinson</a> </td><td> BD-309.306 </td><td></td></tr><tr><td> 2 &quot;x 2&quot; tül pedleri </td><td> Med101store.com </td><td> SKU 2208 </td><td></td></tr><tr><td> 24G ¾ &quot;Angiocath IV kateter </td><td> <a href="http://www.bd.com/" target="_blank">Becton Dickinson</a> </td><td> BD-381.112 </td><td></td></tr><tr><td> 26G ½ &quot;insülin iğnesi </td><td>sıska &quot;&gt; Becton Dickinson </td><td> BD-305.111 </td><td></td></tr><tr><td> Adenozin </td><td> <a href="http://www.sigmaaldrich.com/" target="_blank">Sigma</a> </td><td> A9251 </td><td> 1g / Vazodilatasyon Tampon PBS L (Papaverin ile) </td></tr><tr><td> Açılı Graefe Forseps </td><td> <a href="http://www.finescience.com/" target="_blank">Güzel Bilim Araçları</a> </td><td> 11052-10 </td><td></td></tr><tr><td> Pamuk uçlu aplikatörler: 6 &quot;non-steril </td><td> <a href="http://www.cardinal.com/" target="_blank">Cardinal Health</a> </td><td> C15055-006 </td><td></td></tr><tr><td> Kavisli Cerrahi Makas </td><td> <a href="http://www.finescience.com/" target="_blank">Güzel Bilim Araçları</a> </td><td> 14085-09 </td><td></td></tr><tr><td> Kesme stereoskop ve ışık kaynağı </td><td> <a href="http://www.nikon.com/" target="_blank">Nikon</a> </td><td> NA </td><td> NA </td></tr><tr><td> Kesme Tepsi, 11,5 x 7,5 inç </td><td> <a href="http://www.coleparmer.com/" target="_blank">Cole-Parmer</a> </td><td> YO-10.915-12 </td><td> Fareyi aşağı iğnelemeyi için tar ile Dolu </td></tr><tr><td> Güzel Kavisli Forseps </td><td> <a href="http://www.aesculapusa.com/" target="_blank">Aesculap</a> </td><td> FD281R </td><td> Gereken iki </td></tr><tr><td> Heparin, 5000 U / ml stok </td><td> <a href="http://www.apppharma.com/" target="_blank">APP İlaç LLC</a> </td><td> NDC 63323-047-10 </td><td> Su 1:100 </td></tr><tr><td> KCl </td><td> <a href="http://www.fishersci.com/" target="_blank">Balıkçı</a> </td><td> P217 </td><td> H2O Doymuş çözelti </td></tr><tr><td> Ketamin (Ketaset), 100 mg / ml stok </td><td> Fort Dodge, Overland Park, KS, ABD </td><td> NDC 0856-2013-01 </td><td> % 0.9 &#39;s Xylazine ile 130 mg / kg vücut ağırlığı olarak Karışıkdizilmek </td></tr><tr><td> Microfil </td><td> <a href="http://www.flowtech-inc.com/" target="_blank">Akış Tech</a> </td><td> MV-122 (sarı). Diğer renk seçenekleri de mevcuttur. </td><td> Sertleştirici hacim itibariyle% 10 ile, 1:1 karıştırın. Iyi karışık olduğundan emin olmak için sadece enjeksiyon ve vorteks önce hazırlayın </td></tr><tr><td> Steril olmayan Dikiş: 6-0, örgü ipek </td><td> <a href="http://www.harvardapparatus.com/" target="_blank">Harvard Apparatus</a> </td><td> 723287 </td><td></td></tr><tr><td> Papaverin </td><td> <a href="http://www.americanregent.com/" target="_blank">Amerikan Regent A.Ş.</a> </td><td> NDC 0517-4010-01 </td><td> 4 mg / Vazodilatasyon Tampon PBS L (Adenozin ile) </td></tr><tr><td> Paraformaldehyde </td><td> <a href="http://www.sigmaaldrich.com/" target="_blank">Sigma</a> </td><td> P6148 </td><td> 4% solüsyon olarak hazırlandı </td></tr><tr><td> Perfüzyon Aparatı </td><td></td><td></td><td> Bakınız, şekil 2 </td></tr><tr><td> Bahar Makas </td><td> <a href="http://www.finescience.com/" target="_blank">Güzel Bilim Araçları</a> </td><td> 15018-10 </td><td></td></tr><tr><td> Xylazine (Anased), 20 mg / gl stok </td><td> <a href="http://www.lloydlab.com/" target="_blank">Lloyd Labs</a> </td><td> ADA # 139-236 </td><td> % 0.9 tuzlu su içinde Ketamin ile 8,8 mg / kg vücut ağırlığı gibi karıştırılabilir </td></tr></tbody></table>

retrograde perfusion and filling of mouse coronary vasculature as preparation for micro computed tomography imaging

Damarların görüntülenmesi birçok hastalıktan anlamak için giderek önem kazanmaktadır. Çeşitli teknikler görüntüleme damar için mevcut olmakla birlikte, birkaç küçük gemilere 1,2 içeren bir kararın uzanan bir bütün olarak vasküler ağ görselleştirmek edebiliyoruz. Ayrıca, birçok vasküler döküm teknikleri örnek 3-5 daha fazla analiz önlenmesi, çevre doku yok. Bu konularda circumvents Bir yöntem mikro-Bilgisayarlı Tomografi (μCT) &#39;dir. μCT görüntüleme, kararlar &lt;10 mikron tarama damar ağının 3D rekonstrüksiyon üretme yeteneğine sahip olduğunu ve (örneğin, histoloji ve morfometri) sonraki analiz için sağlam 6-11 doku bırakır olabilir. Bununla birlikte, ex vivo μCT yöntemlerle görüntüleme damarlarının damarlarının radyoopak bileşiği ile doldurulmuş olması gerekir. Bunun gibi, μCT görüntüleme tarafından üretilen damarların doğru temsili üzerine olurludurgemilerin güvenilir ve tam dolum. Bu protokol, biz μCT görüntüleme için hazırlık fare koroner damarları doldurmak için bir teknik açıklar. İki hakim teknikler koroner damar doldurmak için var: Bir Langendorff perfüzyon sistemi 15-17 ile kanülasyon ve retrograd aort perfüzyon (veya arkus kapalı bir şube) 12-14 veya ex vivo ile in vivo. Burada özellikle tüm damarlarda dolum sağlamak için tasarlanmıştır vivo aort kanülasyon yöntemi bir tarif. Bütün kılcal gibi vasküler ağın, arter ve ven her iki tarafında, doldurmak için küçük damarlarda serpmek için Microfil denilen düşük viskoziteli radyopak bileşiğin kullanımı. Kaplar basınçlı bir perfüzyon sistemi kullanılarak tamponu ile perfüze ve sonra Microfil ile doldurulur. Bu Microfil küçük yüksek direnç damarları doldurur sağlamak için, büyük dalları emanatin ligatekoroner arterler içine Microfil yönlendirir aorta, gelen g. Bir kez dolum bazı gemilerin Microfil silip atacak gelen kalp dokusunun elastik yapısı önlemek için, tam, biz hemen doldurduktan sonra erişilebilir büyük damar çıkış noktaları ligate. Bu nedenle, bizim teknik komple koroner damar ağı görselleştirme sağlayarak, dolgu ajanı tam dolum ve maksimum kalıcılık için optimize edilmiştir - arterlerin, kılcal damarları ve damarların hem.

Watch this Scientific Journal Video about Retrograde Perfusion and Filling of Mouse Coronary Vasculature as Preparation for Micro Computed Tomography Imaging at JoVE.com

Retrograde Perfusion and Filling of Mouse Coronary Vasculature as Preparation for Micro Computed Tomography Imaging

Koroner damarların görüntülenmesi kardiyovasküler hastalıkların bizim anlayışın geliştirilmesi için çok önemlidir. Burada mikro-Bilgisayarlı Tomografi (μCT) görüntüleme için hazırlık, bir radyoopak silikon kauçuk (Microfil) ile sıçan koroner damar perfüze için bir yöntem açıklanmaktadır.

Mikro Bilgisayarlı Tomografi Görüntüleme için hazırlık olarak retrograd perfüzyon ve Fare Koroner damarları doldurulması

Visualization of the vasculature is becoming increasingly important for understanding many different disease states. While several techniques exist for ...

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