Name: 3-dimensional resin casting and imaging of mouse portal vein or intrahepatic bile duct system
Uploaded: 2012-10-25T13:00:00
Description: Watch this Scientific Journal Video about 3-Dimensional Resin Casting and Imaging of Mouse Portal Vein or Intrahepatic Bile Duct System at JoVE.com

Dit werk werd ondersteund door subsidies van de National Institutes of Health (NIH) voor SSH (R01-DK078640), van het Howard Hughes Medical Institute (HHMI) door de HHMI / Vanderbilt University Certificate Program in Molecular Medicine aan TJW (GRDOT56006779), de Vanderbilt Diabetes Research en Training Center (P30-DK020593) en de Vanderbilt Digestive Disease Research Center (P30-DK058404) het verstrekken van Core Services.

1. Bereid canule <ol><li> Verwarm een ​​1-inch lange sectie van PE10 slang met uw vingertoppen en zo uitrekken dat de slang dun wordt. Let op: de grootte van het vat of kanaal te worden voorzien van een canule zal de mate van uitrekken nodig vast te stellen. De canule moet goed gespannen voor galgang werpt, maar kan alleen worden verlangd tot matig worden uitgerekt om te passen binnen het grotere poortader. </li><li> Snijd de gestrekte buis bij een diagonaal een afgeschuinde punt genereren. </li><l...

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We hebben beschreven specifieke voorbeelden hoe de poortader en intrahepatische galkanaal systemen van de lever kan worden gegoten, maar deze techniek kan worden toegepast op vrijwel elk ander ductaal of vasculaire systeem met kleine aanpassingen. Eerder werk heeft aangetoond dat het mogelijk deze techniek in verschillende soorten, waaronder muis 1,2,7-9, eendje 10,11, 12,13 konijn, hond 14 en varkens 15, en in veel organen, zoals nasale gland 14, hart 16,</sup...

<table border="1"><tbody><tr><td> Naam van het reagens </td><td> Vennootschap </td><td> Catalogusnummer </td><td> Reacties </td></tr><tr><td> PE10 polyethyleen buis </td><td> Fisher Scientific </td><td> 1417012P </td><td></td></tr><tr><td> 5-0 chirurgische zwart gevlochten zijde </td><td> Roboz Chirurgische </td><td> SUT-15-1 </td><td></td></tr><tr><td> Steriject 32 G x 13 mm naald </td><td> Air-tite </td><td> TSK3213 </td><td></td></tr><tr><td> Lente schaar </td><td> Fine Science Tools </td><td> 15.000-08 </td><td></td></tr><tr><td> Nummer 5 tangen </td><td> Fine Science Tools </td><td> 91.150-20 </td><td></td></tr><tr><td> Mercox II kit </td><td> Ladd Onderzoek </td><td> 21247 </td><td></td></tr><tr><td> Benzylalcohol </td><td> Fisher Scientific </td><td> 1816-04 </td><td> Olleen nodig zijn voor BABB-clearing </td></tr><tr><td> Benzylbenzoaat </td><td> MP Biomedicals, LLC </td><td> 154839 </td><td> Alleen vereist voor BABB-clearing </td></tr><tr><td> Fosfaat-gebufferde zoutoplossing (PBS) </td><td></td><td></td><td></td></tr><tr><td> Boetseerklei </td><td></td><td></td><td></td></tr><tr><td> Schaal </td><td></td><td></td><td></td></tr><tr><td> Laboratorium schaar </td><td></td><td></td><td></td></tr><tr><td> 15 ml cap polypropyleenbuizen </td><td></td><td></td><td></td></tr><tr><td> 4% paraformaldahyde </td><td></td><td></td><td></td></tr><tr><td> 15% kaliumhydroxide (KOH) </td><td></td><td></td><td> Alleen vereist voor maceratie </td></tr><tr><td> Scheermesje </td><td></td><td></td><td></td></tr><tr><td> 100% methanol </td><td></td><td></td><td></td></tr><tr><td> 3 ml luer lock spuit </td><td></td><td></td><td></td></tr></tbody></table>

3-dimensional resin casting and imaging of mouse portal vein or intrahepatic bile duct system

In organen de juiste architectuur van vasculaire en ductale structuren onmisbaar voor een goede fysiologische functie en de vorming en instandhouding van deze structuren is een sterk gereguleerd proces. De analyse van deze complexe, 3-dimensionale structuren sterk afhankelijk of 2-dimensionale onderzoek punt of kleurstof injectie studies. Deze technieken zijn echter niet in staat om een ​​volledige en kwantificeerbare weergave van de ductale of vasculaire structuren zij moeten ophelderen verschaffen. Ook de aard van 3-dimensionale kunsthars worpen genereert een permanente snapshot van het systeem en is een nieuwe en zeer bruikbare techniek voor het visualiseren en kwantificeren van 3-dimensionale structuren en netwerken. Een cruciaal voordeel van het hars gieten systeem is de mogelijkheid om de intacte en aangesloten, of communiceren, structuur van een bloedvat of kanaal te bepalen. De structuur van vasculaire en ductal netwerken zijn cruciaal voor orgaanfunctie en deze techniek heeft de potentie om studie van vasculaire en ductale netwerken steun op verschillende manieren. Hars gieten kan worden gebruikt om normale morfologie en functionaliteit van een luminale structuur analyseren, morfogenetische ontwikkeling veranderingen en ontdekken morfologische verschillen in weefselarchitectuur tussen normale en ziektetoestanden. Eerder werk heeft gebruikt kunstharsproducten te bestuderen, bijvoorbeeld, architectonische en functionele gebreken in de muis intrahepatische galwegen systeem dat niet weerspiegeld in 2-dimensionale analyse van de structuur 1,2, veranderingen in de hersenen vasculatuur van de ziekte van Alzheimer een muismodel 3 , poortader afwijkingen in portaal hypertensieve en cirrose muizen 4, ontwikkelingsstappen in de rat lymfatische rijping tussen onvolwassen en volwassen longen 5, onmiddellijke microvasculaire veranderingen in de rat lever, alvleesklier en nieren onder invloed van de chemische letsel6. Hier presenteren we een methode voor het genereren van een 3-dimensionale hars gegoten van een muis vasculaire of ductaal netwerk, specifiek gericht op de poortader en intrahepatische galwegen. Deze afgietsels kunnen worden gevisualiseerd door het opruimen of macereren het weefsel en kan vervolgens worden geanalyseerd. Deze techniek kan worden toegepast op vrijwel alle vasculaire of ductale systeem en zou rechtstreeks op een studie onderzoek naar de ontwikkeling, functie, onderhoud of letsel van een 3-dimensionaal ductale of vasculaire structuur.

Watch this Scientific Journal Video about 3-Dimensional Resin Casting and Imaging of Mouse Portal Vein or Intrahepatic Bile Duct System at JoVE.com

3-Dimensional Resin Casting and Imaging of Mouse Portal Vein or Intrahepatic Bile Duct System

Werkwijze voor visualiseren en kwantificeren van de 3-dimensionale structuur van muis poortader of intrahepatische galgang beschreven. Deze hars gegoten techniek kan ook worden toegepast op andere ductale of vasculaire systemen en staat<em&gt; In situ</em&gt; Visualisatie of kwantificeren van intacte een systeem communiceert architectuur.

3-Dimensionale kunstharsproducten en beeldvorming van de muis Portal Vein of Intrahepatische Bile Duct System

In  organs, the correct architecture of vascular and ductal structures is  indispensable for proper physiological function, and the formation and  ...

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