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

Wij danken Dr Kelly Stevens voor de eerste proeven van het protocol, Dr Michael Simons, Dr Kip Hauch, en leden van beide van hun labs voor algemene discussie. Dit werk is ondersteuning door NIH subsidies HL087513 en P01 HL094374.

1. Voorbereidingen voor het starten <ol><li> Vul elke zijde van de druk perfusie inrichting met vaatverwijdende buffer (4 mg / L Papaverine + 1 g / L Adenosine in PBS) of 4% paraformaldehyde (PFA) in PBS, respectievelijk. </li><li> Maak een 1/2cc insulinespuit (met een gegoten, 29g ½ &quot;naald) door deze te vullen met 0,1 ml van 1:100 Heparine (5000U/ml voorraad) en het buigen van de naald tot ~ 120 graden hoek met de schuine kant op. Doe de Hetzelfde met een 1 ml spuit (met een 26G ½ &quot;naald) gevuld met 0,3 m...

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Hartweefsel heeft een zeer hoog metabolisme, en vereist dan ook een constante toevoer van voedingsstoffen en zuurstof uit het bloed geleverd door de coronaire vaten. Ziekten van de kransslagaders, die coronaire functie als gevolg van de schepen stenose en verstopping te verminderen, kan leiden tot weefsel hypoxie en ischemie, en zet de getroffen patiënten met risico op myocardinfarct en onherstelbare schade aan de hartspier. Een beter begrip van de zieke toestand van deze schepen is noodzakelijk en van cruciaal belang ...

<html><table border="1"><tbody><tr><td> Naam van het serum </td><td> Vennootschap </td><td> Catalogusnummer </td><td> Reacties </td></tr><tr><td> 1 ml spuiten </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 insuline-injectiespuiten met naalden vast aangesloten 29g ½ &#39; </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; gaasjes </td><td> Med101store.com </td><td> SKU 2208 </td><td></td></tr><tr><td> 24G ¾ &quot;Angiocath IV katheter </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;gauge naalden </td><td>sluik &quot;&gt; Becton Dickinson </td><td> BD-305111 </td><td></td></tr><tr><td> Adenosine </td><td> <a href="http://www.sigmaaldrich.com/" target="_blank">Sigma</a> </td><td> A9251 </td><td> 1 g / L in PBS voor Vasodilatatie Buffer (met Papaverine) </td></tr><tr><td> Schuine 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> Katoen-tip applicators: 6 &quot;niet-steriele </td><td> <a href="http://www.cardinal.com/" target="_blank">Cardinal Health</a> </td><td> C15055-006 </td><td></td></tr><tr><td> Gebogen Chirurgische schaar </td><td> <a href="http://www.finescience.com/" target="_blank">Fine Science Tools</a> </td><td> 14085-09 </td><td></td></tr><tr><td> Het ontleden van stereoscoop en lichtbron </td><td> <a href="http://www.nikon.com/" target="_blank">Nikon</a> </td><td> NA </td><td> NA </td></tr><tr><td> Het ontleden van lade, 11,5 x 7,5 cm </td><td> <a href="http://www.coleparmer.com/" target="_blank">Cole Parmer-</a> </td><td> YO-10915-12 </td><td> Gevuld met bitumen voor overbelasten &#39;van de muis </td></tr><tr><td> Fijn Gebogen tang </td><td> <a href="http://www.aesculapusa.com/" target="_blank">Aesculap</a> </td><td> FD281R </td><td> Er twee nodig </td></tr><tr><td> Heparine, 5000 E / ml bouillon </td><td> <a href="http://www.apppharma.com/" target="_blank">APP Pharmaceuticals LLC</a> </td><td> NDC 63323-047-10 </td><td> 1:100 verdunning in water </td></tr><tr><td> KCl </td><td> <a href="http://www.fishersci.com/" target="_blank">Visser</a> </td><td> P217 </td><td> Verzadigde oplossing in H2O </td></tr><tr><td> Ketamin (Ketaset), 100 mg / ml bouillon </td><td> Fort Dodge, Overland Park, KS, Verenigde Staten </td><td> NDC 0856-2013-01 </td><td> Gemengd als 130 mg / kg lichaamsgewicht, met Xylazine in 0,9% sop een lijn plaatsen </td></tr><tr><td> Microfil </td><td> <a href="http://www.flowtech-inc.com/" target="_blank">Flow Tech</a> </td><td> MV-122 (geel). Andere kleuren zijn ook beschikbaar. </td><td> Meng 01:01 gew met 10 volume% verharder. Bereid vlak voor de injectie, en vortex om ervoor te zorgen is het goed gemengd </td></tr><tr><td> Niet-steriel hechtdraad: 6-0, gevlochten zijde </td><td> <a href="http://www.harvardapparatus.com/" target="_blank">Harvard Apparatuur</a> </td><td> 723287 </td><td></td></tr><tr><td> Papaverine </td><td> <a href="http://www.americanregent.com/" target="_blank">Amerikaanse Regent Inc</a> </td><td> NDC 0517-4010-01 </td><td> 4 mg / L in PBS voor Vasodilatatie Buffer (met adenosine) </td></tr><tr><td> Paraformaldehyde </td><td> <a href="http://www.sigmaaldrich.com/" target="_blank">Sigma</a> </td><td> P6148 </td><td> Bereide 4%-oplossing </td></tr><tr><td> Perfusie Apparatuur </td><td></td><td></td><td> Zie figuur 2 </td></tr><tr><td> Spring Schaar </td><td> <a href="http://www.finescience.com/" target="_blank">Fine Science Tools</a> </td><td> 15018-10 </td><td></td></tr><tr><td> Xylazine (Anased), 20 mg / gl voorraad </td><td> <a href="http://www.lloydlab.com/" target="_blank">Lloyd Labs</a> </td><td> NADA # 139-236 </td><td> Gemengd als 8,8 mg / kg lichaamsgewicht, met Ketamin in 0,9% natriumchloride </td></tr></tbody></table>

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

Visualisatie van het vaatstelsel wordt steeds belangrijker voor het begrijpen van een groot aantal verschillende ziektebeelden. Hoewel diverse technieken bestaan ​​voor beeldvorming vaatstelsel weinig kunnen het vasculaire netwerk als geheel zichtbaar, terwijl de uitbreiding van een resolutie die de kleinere vaartuigen 1,2 bevat. Daarnaast hebben veel vasculaire giettechnieken vernietigen het omliggende weefsel, het voorkomen van verdere analyse van het monster 3-5. Een methode waarin deze aspecten worden omzeilt is micro-Computed Tomography (μCT). μCT beeldvorming kan scannen met een resolutie &lt;10 micron, is geschikt voor het produceren 3D-reconstructies van de vasculaire netwerk, en laat het weefsel intact voor verdere analyse (bijvoorbeeld, histologie en morfometrie) 6-11. Echter, imaging schepen door ex vivo μCT methoden vereist dat de vaten worden gevuld met een contrastmiddel verbinding. Als zodanig is de juiste weergave van de bloedvaten die door μCT beeldvorming is afhankelijk vanbetrouwbare en volledige vulling van de schepen. In dit protocol beschrijven we een techniek voor het vullen van de muis coronaire vaten ter voorbereiding van μCT beeldvorming. Twee overheersen technieken bestaan ​​voor het vullen van de coronaire bloedvaten: in vivo via infusen en retrograde perfusie van de aorta (of een tak van de aortaboog) 12-14, of ex vivo via een Langendorff perfusiesysteem 15-17. Hier beschrijven we een in-vivo aorta cannulatie methode die speciaal is ontworpen om ervoor te zorgen het vullen van alle schepen. We gebruiken lage viscositeit radiopake verbinding genoemd Microfil die wordt doorstroomd door de kleinste schepen de capillairen, evenals zowel de arteriële en veneuze zijden van het vasculaire op te vullen. Schepen worden geperfuseerd met buffer onder druk wordt perfusie-systeem, en vervolgens gevuld met Microfil. Om ervoor te zorgen dat Microfil vult de kleine hogere weerstand schepen, we afbinden van de grote takken emanating van de aorta, die het Microfil afleidt in de kransslagaders. Na het vullen is voltooid, de elastische aard van hartweefsel voorkomen knijpen Microfil uit van enkele schepen, we ligeren toegankelijk belangrijke vasculaire exit-punten onmiddellijk na het vullen. Daarom wordt onze techniek geoptimaliseerd voor volledige vulling en een maximaal behoud van het vulmiddel, zodat visualisatie van de volledige coronaire vasculaire netwerk - slagaders, haarvaten en aders gelijk.

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

Visualisatie van de coronaire bloedvaten is van cruciaal belang voor het bevorderen van onze kennis van hart-en vaatziekten. Hier beschrijven we een methode voor perfuseren muizen coronaire bloedvaten met een contrastmiddel siliconen rubber (Microfil), ter voorbereiding van micro-Computed Tomography (μCT) beeldvorming.

Retrograde Perfusie en vullen van Muis coronaire bloedvaten als voorbereiding op 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

Segmentatie en waardering van de FAT-volumes in muriene modellen Obesitas Met behulp van X-stralen Computer Tomografie

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

Osmotische Drug Delivery voor ischemische achterpoten en perfusie van vaatstelsel met Microfil voor Micro-Computed Tomography 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

Beoordeling van de levensvatbaarheid van menselijk vet Injectie in Naakt Muizen met 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

Met behulp van micro-computertomografie voor de evaluatie van de ontwikkeling van tumoren en follow-up van de respons op de behandeling in een muismodel van longkanker

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

Gecombineerde nabij-infrarood Fluorescent Imaging en Micro-computertomografie voor het direct visualiseren Cerebral Thromboemboli

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

Identificeren van verkalking van de kransslagader op berekend niet-gated tomografie Scans

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

Continue bloedafname in kleine dierlijke positron emissie tomografie/computertomografie maakt de meting van de arteriële ingangsfunctie mogelijk

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

Weefselvoorbereidingstechnieken voor contrastverbeterde microcomputertomografie beeldvorming van grote hartmodellen bij zoogdieren met chronische ziekte

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: Double Resin Casting gevolgd door Micro-Computed Tomography voor 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

Monstervoorbereiding voor op computertomografie gebaseerde driedimensionale visualisatie van murine achterpootvaten

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