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

हम प्रोटोकॉल, डॉ. माइकल सिमंस, डा. रात बिताने का स्थान Hauch के, और सामान्य चर्चा के लिए उनके प्रयोगशालाओं में दोनों के सदस्यों के प्रारंभिक परीक्षणों के लिए धन्यवाद डॉ. केली स्टीवंस. इस काम एनआईएच HL087513 और HL094374 P01 अनुदान द्वारा समर्थन है.

1. शुरू करने से पहले तैयारी <ol><li> Vasodilator (4mg / एल Papaverine + 1g / एल पीबीएस में एक यौगिक जो न्यूक्लिइक अम्ल से प्राप्त होता है जो शर्करा तथा एडीनाइन से बना होता है, डी - राइबोज) के पीबीएस में या 4% Paraformaldehyde बफर (पीएफए) के साथ दबा?...

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हृदय ऊतक के एक बहुत ही उच्च चयापचय की मांग है, और इसलिए कोरोनरी vasculature के द्वारा दिया खून से पोषक तत्वों और ऑक्सीजन की लगातार आपूर्ति की आवश्यकता है. कोरोनरी वाहिकाओं, जो कोरोनरी पोत एक प्रकार का रोग और रुक...

<html><table border="1"><tbody><tr><td> अभिकर्मक का नाम </td><td> कंपनी </td><td> सूचीपत्र संख्या </td><td> टिप्पणियाँ </td></tr><tr><td> 1 मिलीलीटर सीरिंज </td><td> <a href="http://www.bd.com/" target="_blank">Becton डिकिंसन</a> </td><td> BD-309,602 </td><td></td></tr><tr><td> स्थायी रूप से संलग्न 29G साढ़े सुई के साथ 1/2cc इंसुलिन सिरिंजों </td><td> <a href="http://www.bd.com/" target="_blank">Becton डिकिंसन</a> </td><td> BD-309,306 </td><td></td></tr><tr><td> 2 धुंध &quot;x 2&quot; पैड </td><td> Med101store.com </td><td> 2208 .... </td><td></td></tr><tr><td> 24g ¾ &quot;Angiocath चतुर्थ कैथेटर </td><td> <a href="http://www.bd.com/" target="_blank">Becton डिकिंसन</a> </td><td> BD-381,112 </td><td></td></tr><tr><td> 26G साढ़े &quot;गेज सुई </td><td>दुबला&gt; Becton डिकिंसन </td><td> BD-305,111 </td><td></td></tr><tr><td> एक यौगिक जो न्यूक्लिइक अम्ल से प्राप्त होता है जो शर्करा तथा एडीनाइन से बना होता है, डी - राइबोज </td><td> <a href="http://www.sigmaaldrich.com/" target="_blank">सिग्मा</a> </td><td> A9251 </td><td> / 1g, पीबीएस में Vasodilation बफर के लिए एल (Papaverine साथ) </td></tr><tr><td> Angled Graefe संदंश </td><td> <a href="http://www.finescience.com/" target="_blank">ठीक विज्ञान उपकरण</a> </td><td> 11052-10 </td><td></td></tr><tr><td> कपास इत्तला दे दी applicators: 6 &quot;गैर बाँझ </td><td> <a href="http://www.cardinal.com/" target="_blank">प्रमुख स्वास्थ्य</a> </td><td> C15055-006 </td><td></td></tr><tr><td> घुमावदार सर्जिकल कैंची </td><td> <a href="http://www.finescience.com/" target="_blank">ठीक विज्ञान उपकरण</a> </td><td> 14085-09 </td><td></td></tr><tr><td> विदारक त्रिविमदर्शी और प्रकाश स्रोत </td><td> <a href="http://www.nikon.com/" target="_blank">Nikon है</a> </td><td> एनए </td><td> एनए </td></tr><tr><td> विदारक ट्रे, 11.5 x 7.5 इंच </td><td> <a href="http://www.coleparmer.com/" target="_blank">कोल - परमार</a> </td><td> यो - १०,९१५ के-12 </td><td> नीचे माउस लगाए के लिए राल से भरा हुआ </td></tr><tr><td> ठीक घुमावदार संदंश </td><td> <a href="http://www.aesculapusa.com/" target="_blank">Aesculap</a> </td><td> FD281R </td><td> दो की आवश्यकता है </td></tr><tr><td> हेपरिन, 5000 शेयर यू / एमएल </td><td> <a href="http://www.apppharma.com/" target="_blank">एपीपी फार्मास्यूटिकल्स LLC</a> </td><td> 63323-047-10 एनडीसी </td><td> पानी में 1:100 मन्दन </td></tr><tr><td> KCl </td><td> <a href="http://www.fishersci.com/" target="_blank">मछुआ</a> </td><td> P217 </td><td> H2O में संतृप्त समाधान </td></tr><tr><td> (Ketaset) ketamin, 100 स्टॉक मिलीग्राम / एमएल </td><td> किले Dodge, ओवरलैंड पार्क, के एस, संयुक्त राज्य अमेरिका </td><td> 0856-2013-01 एनडीसी </td><td> 130 शरीर मिलीग्राम / किग्रा वजन के रूप में 0.9% में Xylazine साथ, मिश्रितपंक्ति में करना </td></tr><tr><td> MICROFIL </td><td> <a href="http://www.flowtech-inc.com/" target="_blank">प्रवाह टेक</a> </td><td> MV - 122 (पीला). अन्य रंग विकल्प भी उपलब्ध हैं. </td><td> वजन द्वारा 1:01 इलाज एजेंट की मात्रा द्वारा 10% के साथ, मिश्रण. , इंजेक्शन और भंवर से पहले तैयार करने के लिए सुनिश्चित करें के लिए यह अच्छी तरह से मिश्रित है </td></tr><tr><td> गैर बाँझ सीवन: 6-0, लट में रेशम </td><td> <a href="http://www.harvardapparatus.com/" target="_blank">हार्वर्ड उपकरण</a> </td><td> 723,287 </td><td></td></tr><tr><td> Papaverine </td><td> <a href="http://www.americanregent.com/" target="_blank">अमेरिकी रीजेंट इंक</a> </td><td> 0517-4010-01 एनडीसी </td><td> / 4mg, पीबीएस में Vasodilation बफर के लिए एल (Adenosine साथ) </td></tr><tr><td> Paraformaldehyde </td><td> <a href="http://www.sigmaaldrich.com/" target="_blank">सिग्मा</a> </td><td> P6148 </td><td> 4% के समाधान के रूप में तैयार </td></tr><tr><td> छिड़काव उपकरण </td><td></td><td></td><td> चित्र 2 देखें </td></tr><tr><td> वसंत कैंची </td><td> <a href="http://www.finescience.com/" target="_blank">ठीक विज्ञान उपकरण</a> </td><td> 15018-10 </td><td></td></tr><tr><td> (Anased) xylazine, 20 मिलीग्राम / जीएल स्टॉक </td><td> <a href="http://www.lloydlab.com/" target="_blank">लॉयड लैब्स</a> </td><td> 139-236 # नाडा </td><td> 8.8 मिलीग्राम / किग्रा Ketamin साथ शरीर, 0.9% खारा में वजन के रूप में मिश्रित </td></tr></tbody></table>

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

Visualization of the vasculature is becoming increasingly important for understanding many different disease states. While several techniques exist for imaging vasculature, few are able to visualize the vascular network as a whole while extending to a resolution that includes the smaller vessels1,2. Additionally, many vascular casting techniques destroy the surrounding tissue, preventing further analysis of the sample3-5. One method which circumvents these issues is micro-Computed Tomography (&mu;CT). &mu;CT imaging can scan at resolutions &lt;10 microns, is capable of producing 3D reconstructions of the vascular network, and leaves the tissue intact for subsequent analysis (e.g., histology and morphometry)6-11. However, imaging vessels by ex vivo &mu;CT methods requires that the vessels be filled with a radiopaque compound. As such, the accurate representation of vasculature produced by &mu;CT imaging is contingent upon reliable and complete filling of the vessels. In this protocol, we describe a technique for filling mouse coronary vessels in preparation for &mu;CT imaging.
Two predominate techniques exist for filling the coronary vasculature: in vivo via cannulation and retrograde perfusion of the aorta (or a branch off the aortic arch) 12-14, or ex vivo via a Langendorff perfusion system 15-17. Here we describe an in vivo aortic cannulation method which has been specifically designed to ensure filling of all vessels. We use a low viscosity radiopaque compound called Microfil which can perfuse through the smallest vessels to fill all the capillaries, as well as both the arterial and venous sides of the vascular network. Vessels are perfused with buffer using a pressurized perfusion system, and then filled with Microfil. To ensure that Microfil fills the small higher resistance vessels, we ligate the large branches emanating from the aorta, which diverts the Microfil into the coronaries. Once filling is complete, to prevent the elastic nature of cardiac tissue from squeezing Microfil out of some vessels, we ligate accessible major vascular exit points immediately after filling. Therefore, our technique is optimized for complete filling and maximum retention of the filling agent, enabling visualization of the complete coronary vascular network &#x2013; arteries, capillaries, and veins alike.

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

कोरोनरी वाहिकाओं के विज़ुअलाइज़ेशन हृदय रोगों के बारे में हमारी समझ को आगे बढ़ाने के लिए महत्वपूर्ण है. यहाँ हम एक radiopaque सिलिकॉन रबर (MICROFIL) के साथ सूक्ष्म गणना टोमोग्राफी इमेजिंग (μCT) के लिए तैयारी में murine कोरोनरी वाहिका संरचना, perfusing के लिए एक विधि का वर्णन.

प्रतिगामी और सूक्ष्म गणना टोमोग्राफी इमेजिंग के लिए तैयारी के रूप में परफ्यूज़न माउस कोरोनरी vasculature का भरना

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

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