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 tackar Dr Kelly Stevens för de första försöken av protokollet, Dr Michael Simons, Dr Kip Hauch, och medlemmar av båda sina labb för allmän diskussion. Detta arbete är stöd från NIH bidrag HL087513 och P01 HL094374.

1. Förberedelser innan du börjar <ol><li> Fylla varje sida av det tryck perfusionsapparat med Kärlvidgande medel-buffert (4 mg / L Papaverin + 1 g / L Adenosin i PBS) eller 4% paraformaldehyd (PFA) i PBS, respektive. </li><li> Förbered en 1/2cc insulinspruta (med en permanent fastsatt 29G ½ &quot;nål) genom att fylla den med 0,1 ml 1:100 Heparin (5000E/ml materiel) och böja nålen till ~ 120 graders vinkel med avfasningen uppåt. Gör det samma sak med en 1 ml spruta (med en 26G ½ &quot;nål) fylld med 0,3 ml ...

<ol>
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Hjärtvävnad har en mycket hög metabolisk efterfrågan, och därför kräver en konstant tillförsel av näringsämnen och syre från blodet levereras av kranskärlen. Sjukdomar i hjärtats kranskärl, vilket minskar koronar funktion på grund av fartygets stenos och blockering, kan leda till hypoxi och ischemi, och sätta drabbade patienter med risk för hjärtinfarkt och irreparabel skada på hjärtmuskeln. En bättre förståelse av den sjuka tillståndet för dessa fartyg är nödvändigt och avgörande för vår ...

<html><table border="1"><tbody><tr><td> Namnet på reagens </td><td> Företaget </td><td> Katalognummer </td><td> Kommentarer </td></tr><tr><td> 1 ml sprutor </td><td> <a href="http://www.bd.com/" target="_blank">Becton Dickinson</a> </td><td> BD-309.602 </td><td></td></tr><tr><td> 1/2cc insulinsprutor med permanent anbringade 29G ½ &quot;nålar </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; Gauze dynor </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;nålar </td><td>stripigt &quot;&gt; Becton Dickinson </td><td> BD-305.111 </td><td></td></tr><tr><td> Adenosin </td><td> <a href="http://www.sigmaaldrich.com/" target="_blank">Sigma</a> </td><td> A9251 </td><td> 1 g / L i PBS för Vasodilatation buffert (med papaverin) </td></tr><tr><td> Vinklade Graefe Tång </td><td> <a href="http://www.finescience.com/" target="_blank">Fina Science Tools</a> </td><td> 11.052-10 </td><td></td></tr><tr><td> Bomull spets applikatorer: 6 &quot;icke-steril </td><td> <a href="http://www.cardinal.com/" target="_blank">Cardinal Health</a> </td><td> C15055-006 </td><td></td></tr><tr><td> Böjda Kirurgiska Saxar </td><td> <a href="http://www.finescience.com/" target="_blank">Fina Science Tools</a> </td><td> 14.085-09 </td><td></td></tr><tr><td> Dissekera stereoskop och ljuskälla </td><td> <a href="http://www.nikon.com/" target="_blank">Nikon</a> </td><td> NA </td><td> NA </td></tr><tr><td> Dissekera fack, 11,5 x 7,5 inches </td><td> <a href="http://www.coleparmer.com/" target="_blank">Cole-Parmer</a> </td><td> YO-10.915-12 </td><td> Fylld med bitumen för sätter ned musen </td></tr><tr><td> Fina Böjda Tång </td><td> <a href="http://www.aesculapusa.com/" target="_blank">Aesculap</a> </td><td> FD281R </td><td> Behöver två </td></tr><tr><td> Heparin, 5000 U / ml stamlösning </td><td> <a href="http://www.apppharma.com/" target="_blank">APP Läkemedel LLC</a> </td><td> NDC 63323-047-10 </td><td> 1:100 utspädning i vatten </td></tr><tr><td> KCl </td><td> <a href="http://www.fishersci.com/" target="_blank">Fisher</a> </td><td> P217 </td><td> Mättad lösning 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> Blandades som 130 mg / kg kroppsvikt, med xylazin i 0,9% saline </td></tr><tr><td> Mikrofilter </td><td> <a href="http://www.flowtech-inc.com/" target="_blank">Flow Tech</a> </td><td> MV-122 (gul). Andra färgalternativ finns också. </td><td> Blanda 1:1 i vikt, med 10 volym-% av härdningsmedlet. Bered omedelbart före injektion och virvel för att säkerställa att den är väl blandat </td></tr><tr><td> Icke-sterila Suture: 6-0, flätad silke </td><td> <a href="http://www.harvardapparatus.com/" target="_blank">Harvard Apparatus</a> </td><td> 723.287 </td><td></td></tr><tr><td> Papaverin </td><td> <a href="http://www.americanregent.com/" target="_blank">Amerikanska Regent Inc.</a> </td><td> NDC 0517-4010-01 </td><td> 4 mg / L i PBS för Vasodilatation buffert (med adenosin) </td></tr><tr><td> Paraformaldehyd </td><td> <a href="http://www.sigmaaldrich.com/" target="_blank">Sigma</a> </td><td> P6148 </td><td> Framställd såsom 4% lösning </td></tr><tr><td> Perfusionsapparat </td><td></td><td></td><td> Se figur 2 </td></tr><tr><td> Spring Saxar </td><td> <a href="http://www.finescience.com/" target="_blank">Fina Science Tools</a> </td><td> 15.018-10 </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> Blandad som 8,8 mg / kg kroppsvikt, med ketamin i 0,9% koksaltlösning </td></tr></tbody></table>

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

Visualisering av kärlsystemet blir allt viktigare för att förstå många olika sjukdomstillstånd. Medan flera tekniker finns för avbildning kärlsystemet, några har möjlighet att visualisera det vaskulära nätverket i sin helhet, medan sträcker sig till en resolution som innehåller mindre fartyg 1,2. Dessutom har många vaskulära gjutning förstöra den omgivande vävnaden, vilket förhindrar ytterligare analys av provet 3-5. En metod som kringgår dessa frågor är mikro-datortomografi (μCT). μCT avbildning kan skanna i upplösningar &lt;10 mikrometer, kan producera 3D-rekonstruktioner av det vaskulära nätverket och lämnar vävnaden intakt för senare analys (t.ex. histologi och morfometri) 6-11. Kräver dock avbildning fartyg genom ex vivo μCT metoder som fartygen fyllas med en radiopak förening. Som sådan är den exakta återgivning av kärl som produceras av μCT avbildning knutentillförlitlig och fullständig fyllning av fartygen. I detta protokoll, beskriver vi en teknik för att fylla mus kranskärlen som förberedelse för μCT avbildning. Två dominerande tekniker finns för att fylla kranskärlen: in vivo via kanylering och retrograd perfusion av aortan (eller en gren i aortabågen) 12-14, eller ex vivo genom en Langendorff-perfusion systemet 15-17. Här beskriver vi en in vivo-aorta kanylering metod som är speciellt utformade för att säkerställa fyllning av alla fartyg. Vi använder en låg förening viskositet röntgentät kallas mikrofilter som kan perfundera igenom minsta fartygen för att fylla alla kapillärerna, samt både arteriella och venösa sidan av vaskulära nätverket. Kärlen perfuserades med buffert med användning av en trycksatt perfusionssystem, och fylldes sedan med mikrofilter. För att säkerställa att mikrofilter fyller de små högre motstånd fartyg ligera vi stora grenar emanating från aorta, vilken avleder mikrofilter i kranskärlen. När fyllning är klar, för att förhindra den elastiska natur hjärtvävnad från att klämma mikrofilter av vissa fartyg, underbinda vi tillgängliga större vaskulära uttagspunkter omedelbart efter påfyllning. Därför är vår teknik optimerad för fullständig fyllning och maximal retention av fyllmedel, vilket möjliggör visualisering av hela koronar vaskulära nätverk - artärer, kapillärer och vener lika.

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 kranskärlen är avgörande för att öka kunskaperna om hjärt-kärlsjukdomar. Här beskriver vi en metod för perfusion murin kranskärlen med en röntgentät silikongummi (mikrofilter), som förberedelse för mikro-datortomografi (μCT) avbildning.

Retrograd perfusion och påfyllning av mus kranskärlen som förberedelse för Micro datortomografi 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 och Mätning av FAT-volymer i murina fetma modeller med hjälp av X-ray datortomografi

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 ischemisk bakben och perfusion av kärlsystemet med Microfil för Micro-Datortomografi 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

Bedömning av livskraft Human Fat Injektion i nakna möss med Micro-datortomografi

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

Använda Micro-datortomografi för bedömning av tumörutveckling och uppföljning av behandlingssvar i en musmodell av lungcancer

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

Kombinerad nära infraröda fluorescerande Imaging och Micro-datortomografi för direkt Visualisera 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

Att identifiera kranskärl förkalkning på beräknas icke-gated tomografi skanningar

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 blodprovstagning i små djur positron emissions tomografi/datortomografi möjliggör mätning av arteriell input funktion

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

Vävnadsberedningstekniker för kontrastförbättrad mikrodatortomografiavbildning av stora däggdjurshjärtmodeller med kronisk sjukdom

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

DUCT: Dubbelhartsgjutning följt av mikroberäkningstomografi för 3D-leveranalys

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

Provberedning för datortomografibaserad tredimensionell visualisering av Murine Hind-limb Vessels

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