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

우리는 프로토콜 박사 마이클 Simon 씨의, 닥터 킵의 Hauch, 그리고 일반적인 토론을위한 그들의 실험실의 두 구성원들의 초기 실험 용 박사 켈리 스티븐스 감사드립니다. 이 작품은 NIH 보조금 HL087513 및 P01 HL094374에 의해 지원입니다.

1. 시작하기 전에 준비 <ol><li> 각각 PBS에서 Vasodilator 버퍼 (4mg / L Papaverine + 1g / L은 PBS에 아데노신) 또는 4 % Paraformaldehyde (PFA)로 압력 재관류 장치의 각 측면을 채웁니다. </li><li> 1:100 헤파린의 0.1 ML (5000U/ml 주식)로 작성하고 최대 베벨과 함께 ~ 120도 각도로 바늘을 벤딩하여 1/2cc 인슐린 주사기를 (영구적으로 연결된 29G ½ &quot;바늘로) 준비합니다. 수행 0.3 ML 포화 KCl 용액으로 가득 한 ML의 주사기 (26G ...

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심장 조직은 매우 높은 신진 대사 수요를 가지고 있으며, 따라서 관상 vasculature에 의해 전달되는 혈액의 영양분과 산소의 지속적인 공급이 필요합니다. 선박 협착과 막힘으로 인해 관상 동맥 기능을 감소 관상 혈관, 환자는 조직 hypoxia와 국소 빈혈을 초래할, 그리고 심근 경색과 심장 근육에 치유할 수없는 손상에 대한 위험에 영향을받는 환자를 넣을 수 있습니다. 이러한 혈관의 질병 상태에 대한...

<html><table border="1"><tbody><tr><td> 시약의 이름 </td><td> 회사 </td><td> 카탈로그 번호 </td><td> 댓글 </td></tr><tr><td> 한 ML의 주사기 </td><td> <a href="http://www.bd.com/" target="_blank">Becton 디킨슨</a> </td><td> BD-309602 </td><td></td></tr><tr><td> 영구적으로 연결된 29G ½ &#39;바늘과 1/2cc 인슐린 주사기 </td><td> <a href="http://www.bd.com/" target="_blank">Becton 디킨슨</a> </td><td> BD-309306 </td><td></td></tr><tr><td> 2 &quot;X 2&quot;거즈 패드 </td><td> Med101store.com </td><td> SKU 2,208 </td><td></td></tr><tr><td> 24G 잡았다 &quot;Angiocath IV 카테터 </td><td> <a href="http://www.bd.com/" target="_blank">Becton 디킨슨</a> </td><td> BD-381112 </td><td></td></tr><tr><td> 26G ½ &quot;게이지 바늘 </td><td>여윈 &quot;&gt; Becton 디킨슨 </td><td> BD-305111 </td><td></td></tr><tr><td> 아데노신 </td><td> <a href="http://www.sigmaaldrich.com/" target="_blank">시그마</a> </td><td> A9251 </td><td> 1g / Vasodilation 버퍼를위한 PBS에서 L (Papaverine 포함) </td></tr><tr><td> 각도 Graefe의 집게 </td><td> <a href="http://www.finescience.com/" target="_blank">파인 과학 도구</a> </td><td> 11052-10 </td><td></td></tr><tr><td> 면과 스쳐 applicators : 6 &quot;이외의 멸균 </td><td> <a href="http://www.cardinal.com/" target="_blank">추기경 건강</a> </td><td> C15055-006 </td><td></td></tr><tr><td> 곡선 외과 가위 </td><td> <a href="http://www.finescience.com/" target="_blank">파인 과학 도구</a> </td><td> 14085-09 </td><td></td></tr><tr><td> 해부 입체경 및 광원 </td><td> <a href="http://www.nikon.com/" target="_blank">니콘</a> </td><td> NA </td><td> NA </td></tr><tr><td> 해부 트레이, 11.5 X 7.5 인치 </td><td> <a href="http://www.coleparmer.com/" target="_blank">콜 - Parmer</a> </td><td> 요 - 10915-12 </td><td> 마우스를 아래로 달아 위해 타르로 가득 </td></tr><tr><td> 파인 곡선 집게 </td><td> <a href="http://www.aesculapusa.com/" target="_blank">Aesculap</a> </td><td> FD281R </td><td> 게 2 </td></tr><tr><td> 헤파린 5000 U / ML 주식 </td><td> <a href="http://www.apppharma.com/" target="_blank">APP 제약 LLC</a> </td><td> NDC 63323-047-10 </td><td> 물에 1:100 희석 </td></tr><tr><td> KCl </td><td> <a href="http://www.fishersci.com/" target="_blank">어부</a> </td><td> P217 </td><td> H2O의 포화 용액 </td></tr><tr><td> Ketamin (Ketaset), 100 밀리그램 / ML 주식 </td><td> 포트 닷지, 오버랜드 파크, 캔사스, 미국 </td><td> NDC 0856-2013-01 </td><td> 0.9 % s에 Xylazine과 함께, 130 밀리그램 / kg 체중으로 혼합일직선으로하다 </td></tr><tr><td> Microfil </td><td> <a href="http://www.flowtech-inc.com/" target="_blank">흐름 테크</a> </td><td> MV-122 (노랑). 기타 색 옵션도 사용할 수 있습니다. </td><td> 경화 요원의 부피에 의해 10 %, 중량으로 1:1 섞는다. 그것이 잘 혼합한지 단지 분사하고, 소용돌이 전에 준비 </td></tr><tr><td> 비 멸균 봉합해 : 6 대 0으로 꼰 명주실 </td><td> <a href="http://www.harvardapparatus.com/" target="_blank">하버드 장치</a> </td><td> 723,287 </td><td></td></tr><tr><td> Papaverine </td><td> <a href="http://www.americanregent.com/" target="_blank">미국 리젠트 주식 회사</a> </td><td> NDC 0517-4010-01 </td><td> 4mg / Vasodilation 버퍼를위한 PBS에서 L (아데노신 포함) </td></tr><tr><td> Paraformaldehyde </td><td> <a href="http://www.sigmaaldrich.com/" target="_blank">시그마</a> </td><td> P6148 </td><td> 4 % 용액의 준비 </td></tr><tr><td> 관류 장치 </td><td></td><td></td><td> 그림 2 참조 </td></tr><tr><td> 봄 가위 </td><td> <a href="http://www.finescience.com/" target="_blank">파인 과학 도구</a> </td><td> 15018-10 </td><td></td></tr><tr><td> Xylazine (Anased), 20 밀리그램 / GL 주식 </td><td> <a href="http://www.lloydlab.com/" target="_blank">로이드 실험실</a> </td><td> 나다 # 139-236 </td><td> 0.9 %의 식염수에 Ketamin과 8.8 밀리그램 / kg 체중으로 혼합 </td></tr></tbody></table>

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

vasculature의 시각화는 여러 질병 상태를 이해를 위해 점점 더 중요 해지고 있습니다. 여러 기법 이미징 vasculature을 위해 존재하지만, 몇몇 작은 혈관의 1,2를 포함 해상도로 확대하면서 전체적으로 혈관 네트워크를 시각화 할 수 있습니다. 또한 많은 혈관 주조 기법 예제 3-5의 자세한 분석을 방지, 주변 조직을 파괴. 이러한 문제를 circumvents 한 가지 방법은 마이크로 계산된 Tomography (μCT)입니다. μCT 영상은 해상도 &lt;10 미크론에서 스캔 혈관 네트워크의 3 차원 reconstructions를 생산 할 수 있으며 (예 : 조직학 및 morphometry) 후속 분석을 위해 그대로 6-11을 조직을 떠날 수 있습니다. 그러나 전직 생체내 μCT 방법에 의한 이미징 혈관 혈관이 radiopaque 화합물로 가득해야합니다. 따라서 μCT 이미징 제작한 vasculature의 정확한 표현이 조건으로하고있다혈관의 신뢰성과 완벽한 충전. 이 프로토콜에서는 μCT 이미징을위한 준비 마우스 관상 동맥 혈관을 작성을위한 기법을 설명합니다. 두 predominate 기술은 관상 vasculature 가득 채우는 것은 존재 : Langendorff 재관류 시스템을 통해 15-17 cannulation과 역행 대동맥의 관류 (또는 대동맥 아치 해제 지점) 12-14, 또는 전직 생체내 통해 생체내 인치 여기에서 우리는 특별히 모든 선박에 대해 작성하도록 설계되었습니다 생체내의 대동맥 cannulation 방법에 대해 설명합니다. 우리 모두는 모세 혈관뿐만 아니라, 혈관 네트워크의 동맥과 정맥의 양쪽 모두를 채우기 위해 작은 혈관을 통해 perfuse 수 Microfil라는 낮은 점도의 radiopaque 화합물을 사용합니다. 혈관이 압력 관류 시스템을 사용하여 버퍼로 perfused 후 Microfil으로 가득합니다. 그 Microfil가 작은 높은 저항 혈관을 채우고 확보하기 위해 대규모 지점 emanatin ligatecoronaries로 Microfil을 산란 대동맥에서 g. 일단 충전 일부 표본 Microfil을 짜면 심장 조직의 탄성 특성을 방지하기 위해, 완료 즉시 충전 후 접근이 주요 혈관 출구 포인트를 ligate. 따라서, 우리의 기술은 완전한 관상 혈관 네트워크의 시각화을 사용, 충전 대리인의 완전한 충전과 최대 보존에 최적화되어 있습니다 - 동맥, 모세 혈관과 정맥이 비슷하다.

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

관상 혈관의 시각화는 심장 혈관 질병의 우리의 이해를 증진하기 위해 중요합니다. 여기에서 우리는 마이크로 계산된 Tomography (μCT) 이미징을위한 준비, radiopaque 실리콘 고무 (Microfil)로 murine 관상 vasculature을 perfusing위한 방법을 설명합니다.

마이크로 계산된 Tomography 이미징을위한 준비로서 역행 관류 및 마우스 관상 Vasculature의 작성

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

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