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 til indledende forsøg i protokollen, Dr. Michael Simons, Dr. Kip Hauch, og medlemmer af begge deres laboratorier til generel diskussion. Dette arbejde er støtte fra NIH tilskud HL087513 og P01 HL094374.

1. Forberedelser før start <ol><li> Fylde hver side af trykket perfusion apparatet med vasodilator puffer (4mg / l Papaverin + 1 g / L Adenosin i PBS) eller 4% paraformaldehyd (PFA) i PBS, hhv. </li><li> Forbered en 1/2cc insulinsprøjte (med fastmonteret 29g ½ &quot;nål) ved at fylde den med 0,1 ml 1:100 Heparin (5000U/ml lager) og bøje nålen til ~ 120 graders vinkel med facet op. Gør samme med en 1 ml sprøjte (med en 26G ½ &quot;nål) fyldt med 0,3 ml mættet KCI-opløsning. </li></ol> ...

<ol>
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Hjertevæv har en meget høj metabolisk behov, og derfor kræver en konstant tilførsel af næringsstoffer og oxygen fra blodet leveret af koronar vaskulatur. Sygdomme i koronarkar, som aftager koronar funktion på grund af skibet stenose og blokering, kan føre til hypoxi og iskæmi, og sætte de berørte patienter med risiko for myokardieinfarkt og uoprettelig skade på hjertemusklen. En bedre forståelse af den syge tilstand af disse fartøjer er nødvendig, og afgørende for vores evne til at studere koronarkar er v...

<html><table border="1"><tbody><tr><td> Navn på reagenset </td><td> Firma </td><td> Katalog nummer </td><td> Kommentarer </td></tr><tr><td> 1 ml injektionssprøjter </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 insulininjektionssprøjter med fast anbragte 29g ½ &#39;nåle </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; gazetamponer </td><td> Med101store.com </td><td> SKU 2208 </td><td></td></tr><tr><td> 24G ¾ &quot;angiokateter 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;gauge nåle </td><td>lank &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> 1g / l i PBS i Vasodilatation buffer (med papaverin) </td></tr><tr><td> Vinklede Graefe Tang </td><td> <a href="http://www.finescience.com/" target="_blank">Fine Science Tools</a> </td><td> 11052-10 </td><td></td></tr><tr><td> Bomuld tippes applikatorer: 6 &quot;ikke-steril </td><td> <a href="http://www.cardinal.com/" target="_blank">Cardinal Health</a> </td><td> C15055-006 </td><td></td></tr><tr><td> Buede operationssaks </td><td> <a href="http://www.finescience.com/" target="_blank">Fine Science Tools</a> </td><td> 14085-09 </td><td></td></tr><tr><td> Dissekering stereoskop 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 Bakke, 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> Fyldt med tjære til pinning 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> Har du brug for to </td></tr><tr><td> Heparin, 5000 U / ml stamopløsning </td><td> <a href="http://www.apppharma.com/" target="_blank">APP Pharmaceuticals LLC</a> </td><td> NDC 63323-047-10 </td><td> 1:100 fortynding i vand </td></tr><tr><td> KCl </td><td> <a href="http://www.fishersci.com/" target="_blank">Fisher</a> </td><td> P217 </td><td> Mættet opløsning i H2O </td></tr><tr><td> Ketamin (Ketaset), 100 mg / ml stamopløsning </td><td> Fort Dodge, Overland Park, KS, USA </td><td> NDC 0856-2013-01 </td><td> Blandet med 130 mg / kg legemsvægt, 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 farver er også tilgængelige. </td><td> Blandes 1:1 efter vægt, med 10 vol-% af hærdningsmidlet. Forbered lige før injektion, og vortex for at sikre, at det er godt blandet </td></tr><tr><td> Ikke-sterile 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> 4 mg / L i PBS i Vasodilatation buffer (med adenosin) </td></tr><tr><td> Paraformaldehyd </td><td> <a href="http://www.sigmaaldrich.com/" target="_blank">Sigma</a> </td><td> P6148 </td><td> Fremstillet som 4% opløsning </td></tr><tr><td> Perfusion Apparat </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 Tools</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 med 8,8 mg / kg legemsvægt, med ketamin i 0,9% saltvand </td></tr></tbody></table>

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

Visualisering af vaskulaturen bliver stadig vigtigere for forståelsen af ​​mange forskellige sygdomstilstande. Mens flere teknikker eksisterer for billedbehandling kar, få er i stand til at visualisere det vaskulære netværk som en helhed og samtidig udvide til en resolution, der omfatter mindre skibe 1,2. Desuden er mange vaskulære støbeteknikker ødelægge det omgivende væv, hvilket forhindrer yderligere analyse af prøven 3-5. En metode, der omgås disse spørgsmål, er mikro-Computed Tomography (μCT). μCT billeddannelse kan scanne i opløsninger &lt;10 um, er i stand til at producere 3D rekonstruktion af det vaskulære netværk, og efterlader vævet intakt til efterfølgende analyse (f.eks histologi og morfometri) 6-11. Men, billedbehandling fartøjer ved ex vivo μCT metoder kræver, at fartøjer fyldt med en røntgenfast stof. Som sådan er den nøjagtige gengivelse af vaskulaturen fremstilles ved μCT billeddannelse betingetpålidelige og fuldstændige fyldning af skibene. I denne protokol, beskriver vi en teknik til at fylde mus koronarkar i forberedelse til μCT billeddannelse. To dominerer teknikker eksisterer til fyldning af koronar vaskulaturen: in vivo via en kanyle og retrograd perfusion af aorta (eller en gren af aortabuen) 12-14 eller ex vivo via en Langendorff perfusionssystem 15-17. Her beskriver vi en in vivo aorta kanylering metode, der er specielt designet til at sikre fyldning af alle fartøjer. Vi anvender en lav viskositet røntgenabsorberende stof kaldet Microfil som kan perfundere gennem mindste fartøjer for at fylde hele kapillærer, såvel som både arterielle og venøse side af det vaskulære netværk. Skibe perfunderet med buffer med en tryksat perfusionssystem, og derefter fyldt med Microfil. At sikre, at Microfil fylder mindre højere modstand fartøjer vi ligere de store grene emanating fra aorta, som afleder Microfil ind i blodpropper. Når påfyldning er fuldført, for at forhindre den elastiske beskaffenhed af hjertevæv fra presse Microfil af nogle kar, vi ligere tilgængelige store vaskulære udgangssteder umiddelbart efter fyldning. Derfor er vores teknik optimeret til fuldstændig fyldning og maksimal retention af fyldstof, muliggør visualisering af hele koronar vaskulære netværk - arterier, kapillærer og vener ens.

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 af koronarkar er afgørende for at fremme vores forståelse af hjerte-kar-sygdomme. Her beskriver vi en metode til perfusion murine koronar kar med en røntgenfast silikonegummi (Microfil), som forberedelse til mikro-Computed Tomography (μCT) billeddannelse.

Retrograd perfusion og påfyldning af Mouse koronar kar som forberedelse til Micro Computed Tomography 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 af tykke bind i murine Fedme-modeller vha. X-ray Computed Tomography

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 Iskæmisk bagbensknogler og perfusion af kar med Microfil for Micro-computertomografi 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 af virksomhedens rentabilitet for Human Fat Injektion i nøgne mus med Micro-Computed Tomography

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

Brug Micro-computertomografi for Vurdering af Tumor Udvikling og opfølgning af respons på behandling i en musemodel for lungekræft

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

Kombineret 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

At identificere koronararterie forkalkning på beregnet ikke-gated tomografi scanninger

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 blodprøvetagning i små dyr Positron emission Tomography/computertomografi gør det muligt at måle den arterielle indgangsfunktion

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

Vævsforberedelsesteknikker til kontrastforstærket mikrocomputertomografibilleddannelse af store pattedyrs hjertemodeller med kronisk sygdom

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: Dobbelt harpiksstøbning efterfulgt af mikro-computertomografi til 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

Prøveforberedelse til computertomografibaseret tredimensionel visualisering af 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 ...