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

Vi takker Dr. Kelly Stevens for første utprøving av protokollen, Dr. Michael Simons, Dr. Kip Hauch, og medlemmer av begge sine laboratorier for generell diskusjon. Dette arbeidet er støtte ved NIH tilskudd HL087513 og P01 HL094374.

1. Forberedelser før start <ol><li> Fyll hver side av trykket perfusjons apparat med vasodilator buffer (4mg / L papaverin + 1g / L Adenosin i PBS) eller 4% Paraformaldehyde (PFA) i PBS, henholdsvis. </li><li> Forbered en 1/2cc Insulin sprøyte (med en fast tilkoblet 29G ½ &quot;p) ved å fylle den med 0,1 ml 1:100 Heparin (5000U/ml lager) og bøye nålen til ~ 120 graders vinkel med skråkant opp. Gjør samme med en 1 ml sprøyte (med en 26G ½ &quot;nålen) fylt med 0,3 ml mettet KCl løsning. </li></ol><p class="...

<ol>
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Cardiac vev har en svært høy metabolsk etterspørsel, og derfor krever en konstant tilførsel av næringsstoffer og oksygen fra blodet levert av koronar blodkar. Sykdommer i hjertets fartøy, som reduserer koronar funksjon på grunn av fartøy stenose og blokkering, kan føre til vevshypoksi og iskemi, og sette berørte pasienter med risiko for hjerteinfarkt og uopprettelig skade på hjertemuskelen. En bedre forståelse av sykelig tilstand av disse fartøyene er nødvendig, og kritisk til vår evne til å studere koro...

<html><table border="1"><tbody><tr><td> Navn på reagensen </td><td> Firma </td><td> Katalognummer </td><td> Kommentarer </td></tr><tr><td> 1 ml sprøyter </td><td> <a href="http://www.bd.com/" target="_blank">Becton Dickinson</a> </td><td> BD-309602 </td><td></td></tr><tr><td> 1/2cc insulin sprøyter med fast tilkoblet 29G ½ &#39;nåler </td><td> <a href="http://www.bd.com/" target="_blank">Becton Dickinson</a> </td><td> BD-309306 </td><td></td></tr><tr><td> 2 &quot;x 2&quot; gasbind pads </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-381112 </td><td></td></tr><tr><td> 26G ½ &quot;måle nåler </td><td>mager &quot;&gt; Becton Dickinson </td><td> BD-305111 </td><td></td></tr><tr><td> Adenosin </td><td> <a href="http://www.sigmaaldrich.com/" target="_blank">Sigma</a> </td><td> A9251 </td><td> 1g / L i PBS for Vasodilatasjon buffer (med papaverin) </td></tr><tr><td> Vinklet Græfe tang </td><td> <a href="http://www.finescience.com/" target="_blank">Fine Science Verktøy</a> </td><td> 11052-10 </td><td></td></tr><tr><td> Bomull-tipped applikator: 6 &quot;ikke-sterile </td><td> <a href="http://www.cardinal.com/" target="_blank">Cardinal Health</a> </td><td> C15055-006 </td><td></td></tr><tr><td> Buede Kirurgiske Saks </td><td> <a href="http://www.finescience.com/" target="_blank">Fine Science Verktøy</a> </td><td> 14085-09 </td><td></td></tr><tr><td> Dissekere stereoscope og lyskilde </td><td> <a href="http://www.nikon.com/" target="_blank">Nikon</a> </td><td> NA </td><td> NA </td></tr><tr><td> Dissekere skuff, 11.5 x 7.5 inches </td><td> <a href="http://www.coleparmer.com/" target="_blank">Cole-Parmer</a> </td><td> YO-10915-12 </td><td> Fylt med tjære for låsing ned musen </td></tr><tr><td> Fine Buede tang </td><td> <a href="http://www.aesculapusa.com/" target="_blank">Aesculap</a> </td><td> FD281R </td><td> Trenger to </td></tr><tr><td> Heparin, 5000 U / ml lager </td><td> <a href="http://www.apppharma.com/" target="_blank">APP Pharmaceuticals LLC</a> </td><td> NDC 63323-047-10 </td><td> 1:100 fortynning i vann </td></tr><tr><td> KCl </td><td> <a href="http://www.fishersci.com/" target="_blank">Fisher</a> </td><td> P217 </td><td> Mettet løsning i H2O </td></tr><tr><td> Ketamin (Ketaset), 100 mg / ml lager </td><td> Fort Dodge, Overland Park, KS, USA </td><td> NDC 0856-2013-01 </td><td> Blandet som 130 mg / kg kroppsvekt, med xylazin i 0,9% sAline </td></tr><tr><td> Microfil </td><td> <a href="http://www.flowtech-inc.com/" target="_blank">Flow Tech</a> </td><td> MV-122 (gul). Andre fargealternativer er også tilgjengelig. </td><td> Bland 01:01 av vekt, med 10% av volumet av herder. Forbered like før injeksjon, og virvle å sikre at det er godt blandet </td></tr><tr><td> Ikke-steril Sutur: 6-0, flettet silke </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">American Regent Inc.</a> </td><td> NDC 0517-4010-01 </td><td> 4mg / L i PBS for Vasodilatasjon buffer (med Adenosine) </td></tr><tr><td> Paraformaldehyde </td><td> <a href="http://www.sigmaaldrich.com/" target="_blank">Sigma</a> </td><td> P6148 </td><td> Forberedt som 4% løsning </td></tr><tr><td> Perfusjon Apparatus </td><td></td><td></td><td> Se figur 2 </td></tr><tr><td> Spring Saks </td><td> <a href="http://www.finescience.com/" target="_blank">Fine Science Verktøy</a> </td><td> 15018-10 </td><td></td></tr><tr><td> Xylazin (Anased), 20 mg / gl lager </td><td> <a href="http://www.lloydlab.com/" target="_blank">Lloyd Labs</a> </td><td> NADA # 139-236 </td><td> Blandet som 8,8 mg / kg kroppsvekt, med Ketamin i 0,9% saltløsning </td></tr></tbody></table>

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

Visualisering av blodkar blir stadig viktigere for å forstå mange ulike sykdomstilstander. Mens flere teknikker finnes for bildebehandling blodkar, få er i stand til å visualisere vaskulære nettverket som helhet, mens strekker seg til en oppløsning som inneholder mindre fartøyer 1,2. I tillegg er mange vaskulære kasteteknikker ødelegge det omkringliggende vevet, og hindrer videre analyse av prøven 3-5. En metode som omgår disse problemene er mikro-computertomografi (μCT). μCT bildebehandling kan skanne med en oppløsning &lt;10 mikron, er i stand til å produsere 3D rekonstruksjoner av vaskulær nettverket, og forlater vev inntakt for påfølgende analyse (f.eks histologi og morfometri) 6-11. Krever imidlertid tenkelig fartøy av ex vivo μCT metoder som fartøyene bli fylt med et røntgentett sammensatt. Som sådan er det nøyaktig representasjon av blodkar produsert av μCT tenkelig betingetpålitelig og fullstendig fylling av fartøyene. I denne protokollen, beskriver vi en teknikk for å fylle mus koronar fartøy i forberedelse til μCT bildebehandling. To dominerer teknikker finnes for å fylle koronar blodkar: in vivo via kanylering og retrograd perfusjon av aorta (eller en filial av aortabuen) 12-14, eller ex vivo via en Langendorff perfusjon system 15-17. Her beskriver vi en in vivo aorta kanylering metode som er spesielt utformet for å sikre fylling av alle fartøy. Vi bruker en lav viskositet røntgentett stoff kalt Microfil som kan perfuse gjennom de minste fartøyene til å fylle alle kapillærene, samt både arterielle og venøse sider av vaskulær nettverket. Fartøyene er perfused med buffer ved hjelp av en trykksatt perfusjon system, og deretter fylt med Microfil. For å sikre at Microfil fyller de små høyere motstand fartøy, ligate vi de store grenene emanating fra aorta, som viderekoble Microfil inn i coronaries. Når fylling er ferdig, for å hindre elastisk natur hjertevev fra klemme Microfil ut av noen skip, ligate vi tilgjengelige store vaskulære utganger umiddelbart etter fylling. Derfor er vår teknikk optimalisert for fullstendig utfylling og maksimal oppbevaring av fylling agent, slik visualisering av hele koronar vaskulær nettverk - arterier, kapillærer og vener like.

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

Visualisering av hjertets fartøyene er kritisk til å fremme vår forståelse av hjerte-og karsykdommer. Her beskriver vi en metode for perfusing murine koronar blodkar med et røntgentett silikongummi (Microfil), i forberedelse for mikro-computertomografi (μCT) bildebehandling.

Retrograd perfusjon og Fylling av Mouse Coronary blodkar som forberedelse for Micro Datatomografiscanning Imaging

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

Research

JoVE Journal

Medicine

Segmentation and Measurement of Fat Volumes in Murine Obesity Models Using X-ray Computed Tomography

Segmentering og Måling av fett Volumene i murine Fedme modeller med X-ray Datatomografiscanning

Obesity is associated with increased morbidity and mortality as well as reduced metrics in quality of life.1 Both environmental and genetic factors are ...

Osmotic Drug Delivery to Ischemic Hindlimbs and Perfusion of Vasculature with Microfil for Micro-Computed Tomography Imaging

Osmotisk Drug Delivery til iskemisk bakbena og perfusjon av blodkar med Microfil for Micro-CT Imaging

Preclinical research in animal models of peripheral arterial disease plays a vital role in testing the efficacy of therapeutic agents designed to ...

Assessment of Viability of Human Fat Injection into Nude Mice with Micro-Computed Tomography

Vurdering av Livskraftig of Human Fat Injeksjon i Nude Mus med Micro-Datatomografiscanning

Lipotransfer is a vital tool in the surgeon&#8217;s armamentarium for the treatment of soft tissue deficits of throughout the body. Fat is the ideal soft ...

Using Micro-computed Tomography for the Assessment of Tumor Development and Follow-up of Response to Treatment in a Mouse Model of Lung Cancer

Ved hjelp av Micro-computertomografi for vurdering av tumorutvikling og oppfølging av respons på behandlingen i en musemodell for lungekreft

Lung cancer is the most lethal cancer in the world. Intensive research is ongoing worldwide to identify new therapies for lung cancer. Several mouse ...

Combined Near-infrared Fluorescent Imaging and Micro-computed Tomography for Directly Visualizing Cerebral Thromboemboli

Kombinert Nær-infrarød Fluorescent Imaging og Micro-computertomografi for Direkte Visualisering Cerebral tromboemboli

Direct thrombus imaging visualizes the root cause of thromboembolic infarction. Being able to image thrombus directly allows far better investigation of ...

Identifying Coronary Artery Calcification on Non-gated Computed Tomography Scans

Identifisere koronar forkalkninger på beregnet ikke-gated tomografi skanner

Coronary artery calcification (CAC) provides an objective measure of coronary artery disease and can readily be identified on non-gated computed ...

Continuous Blood Sampling in Small Animal Positron Emission Tomography/Computed Tomography Enables the Measurement of the Arterial Input Function

Kontinuerlig blodprøve i små dyr positron utslipp tomografi/beregnet tomografi muliggjør måling av arteriell input-funksjonen

For quantitative analysis and bio-kinetic modeling of positron emission tomography/computed tomography (PET/CT) data, the determination of the temporal ...

Tissue Preparation Techniques for Contrast-Enhanced Micro Computed Tomography Imaging of Large Mammalian Cardiac Models with Chronic Disease

Vevsforberedelsesteknikker for kontrastforbedret mikroberegnet tomografiavbildning av store pattedyr hjertemodeller med kronisk sykdom

Structural remodeling is a common consequence of chronic pathological stresses imposed on the heart. Understanding the architectural and compositional ...

DUCT: Double Resin Casting followed by Micro-Computed Tomography for 3D Liver Analysis

KANAL: Dobbel harpiksstøping etterfulgt av mikro-beregnet tomografi for 3D leveranalyse

The liver is the biggest internal organ in humans and mice, and high auto-fluorescence presents a significant challenge for assessing the ...

Sample Preparation for Computed Tomography-based Three-dimensional Visualization of Murine Hind-limb Vessels

Prøvepreparering for beregnet tomografibasert tredimensjonal visualisering av Murine Hind-limb-kar

Blood vessels are complex networks with tree-like structures, and vascular networks are essential for maintaining both circulation and maintaining organ ...