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

Questo lavoro è stato sostenuto dalle concessioni dal National Institutes of Health (NIH) a SSH (R01-DK078640), del Howard Hughes Medical Institute (HHMI) attraverso il HHMI / Vanderbilt Certificate Program University di Medicina Molecolare per TJW (GRDOT56006779), la Vanderbilt Diabetes Research and Training Center (P30-DK020593) e la Digestive Disease Research Center Vanderbilt (P30-DK058404) fornendo servizi di base.

1. Preparare Cannula <ol><li> Scaldare un 1-pollice lungo tratto di PE10 tubi con la punta delle dita e allungare in modo che il tubo diventa sottile. Nota: la dimensione del recipiente o condotto da incannulata determinerà il grado di stiramento richiesto. La cannula deve essere ben teso per via biliare getta, ma può essere richiesto solo per essere moderatamente allungato per adattarsi all&#39;interno della vena porta più grande. </li><li> Tagliare il tubo allungato in diagonale per generare una p...

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C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evidence+for+a+unique+elastic+sheath+surrounding+the+vesicular+arteries+of+the+rabbit+urinary+bladder--studies+of+the+microvasculature+with+microscopy+and+vascular+corrosion+casting.">Evidence for a unique elastic sheath surrounding the vesicular arteries of the rabbit urinary bladder--studies of the microvasculature with microscopy and vascular corrosion casting.</a> Anat. Rec. 252, 472-476 (1998).</li><li>Hossler, F. E., Kao, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microvasculature+of+the+urinary+bladder+of+the+dog:+a+study+using+vascular+corrosion+casting.">Microvasculature of the urinary bladder of the dog: a study using vascular corrosion casting.</a> Microsc. Microanal. 13, 220-227 (2007).</li><li>Wischgoll, T., Choy, J. S., Kassab, G. 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E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vascular+Corrosion+Casting:+Review+of+Advantages+and+Limitations+in+the+Application+of+Some+Simple+Quantitative+Methods.">Vascular Corrosion Casting: Review of Advantages and Limitations in the Application of Some Simple Quantitative Methods.</a> Microsc. Microanal. 7, 253-264 (2001).</li><li>Mondy, W. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Micro-CT+of+corrosion+casts+for+use+in+the+computer-aided+design+of+microvasculature.">Micro-CT of corrosion casts for use in the computer-aided design of microvasculature.</a> Tissue Eng. Part C Methods. 15, 729-738 (2009).</li><li>Masyuk, T. V., Ritman, E. L., LaRusso, N. 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Abbiamo descritto esempi specifici di come la vena porta e intraepatiche sistemi dotti biliari del fegato può essere fusa, ma questa tecnica può essere applicata a qualsiasi altro sistema duttale o vascolare con lievi modifiche. Il lavoro precedente ha dimostrato la fattibilità di questa tecnica in più specie, tra cui mouse, 1,2,7-9 anatroccolo 10,11, 12,13 coniglio, cane 14, e maiale 15, e in molti organi, tra cui ghiandola nasale 14, cuore 16, 12,13...

<table border="1"><tbody><tr><td> Nome del reagente </td><td> Azienda </td><td> Numero di catalogo </td><td> Commenti </td></tr><tr><td> PE10 tubi in polietilene </td><td> Fisher Scientific </td><td> 1417012P </td><td></td></tr><tr><td> 5-0 chirurgico di seta nera intrecciata </td><td> Roboz chirurgico </td><td> SUT-15-1 </td><td></td></tr><tr><td> Steriject 32 G x 13 mm ago </td><td> Air-tite </td><td> TSK3213 </td><td></td></tr><tr><td> Primavera forbici </td><td> Strumenti Scienza Belle </td><td> 15000-08 </td><td></td></tr><tr><td> Numero 5 pinza </td><td> Strumenti Scienza Belle </td><td> 91150-20 </td><td></td></tr><tr><td> Mercox II Kit </td><td> Ladd ricerca </td><td> 21247 </td><td></td></tr><tr><td> Alcool benzilico </td><td> Fisher Scientific </td><td> 1816-1804 </td><td> Oolo necessario per BABB-compensazione </td></tr><tr><td> Benzoato di benzile </td><td> MP Biomedicals, LLC </td><td> 154839 </td><td> Richiesto solo per BABB-compensazione </td></tr><tr><td> Tampone fosfato salino (PBS) </td><td></td><td></td><td></td></tr><tr><td> Modellazione di argilla </td><td></td><td></td><td></td></tr><tr><td> Scala </td><td></td><td></td><td></td></tr><tr><td> Laboratorio di forbici </td><td></td><td></td><td></td></tr><tr><td> 15 ml tappo in polipropilene tubi </td><td></td><td></td><td></td></tr><tr><td> 4% paraformaldahyde </td><td></td><td></td><td></td></tr><tr><td> Idrossido di potassio al 15% (KOH) </td><td></td><td></td><td> Richiesto solo per la macerazione </td></tr><tr><td> Lametta </td><td></td><td></td><td></td></tr><tr><td> 100% metanolo </td><td></td><td></td><td></td></tr><tr><td> 3 ml luer lock siringa </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

Di organi, l&#39;architettura corretta delle strutture vascolari e duttale è indispensabile per una corretta funzione fisiologica, e la formazione e il mantenimento di queste strutture è un processo altamente regolato. L&#39;analisi di questi complessi, tre strutture tridimensionali è notevolmente dipendeva sia 2-dimensionale esame in sezione o su studi iniezione tintura. Queste tecniche, tuttavia, non sono in grado di fornire una rappresentazione completa e quantificabile delle strutture duttali o vascolari sono destinati a chiarire. In alternativa, la natura del 3-dimensionale calchi in resina plastica genera un&#39;istantanea permanente del sistema ed è un romanzo e tecnica ampiamente utile per la visualizzazione e la quantificazione tre strutture tridimensionali e reti. Un vantaggio fondamentale del sistema di colata di resina è la capacità di determinare la intatte e collegate, o comunicare, struttura di un vaso sanguigno o condotto. La struttura vascolare e di ducreti mentali sono essenziali per la funzione degli organi, e questa tecnica ha il potenziale per aiutare studio delle reti vascolari e duttale in diversi modi. Colata di resina può essere utilizzata per analizzare la morfologia normale e l&#39;architettura funzionale di una struttura luminale, identificare cambiamenti morfogenetici sviluppo, e scoprire differenze morfologiche dell&#39;architettura tessuti normali tra stati e malattie. Lavoro precedente ha utilizzato resina colata a studiare, per esempio, difetti architettoniche e funzionali nel sistema biliare intraepatica topo condotto che non si sono riflesse in 2-dimensionale analisi della struttura 1,2, alterazioni vascolare cerebrale di un modello di malattia di Alzheimer mouse 3 , anomalie della vena porta nel portale topi ipertesi e cirrotici 4, passi di sviluppo in maturazione ratto linfatico tra polmoni immaturi e adulti 5, immediati cambiamenti microvascolari nel fegato di ratto, del pancreas e del rene in risposta a al danno chimico6. Presentiamo qui un metodo di generazione di un 3-dimensionale di resina di una rete vascolare del mouse o duttale specificamente dedicato alla vena porta e dotto biliare intraepatica. Questi calchi possono essere visualizzati tramite compensazione o macerazione del tessuto e possono quindi essere analizzati. Questa tecnica può essere applicata a qualsiasi sistema vascolare o duttale e sarebbe direttamente applicabile a qualsiasi studio indagare lo sviluppo, la funzione, la manutenzione, o lesioni di un 3-dimensionale struttura duttale o vascolare.

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

Un metodo di visualizzare e quantificare il 3-dimensionale struttura vena porta mouse o dotto biliare intraepatica è descritto. Questa tecnica resina può anche essere applicata ad altri sistemi duttali o vascolare e permette<em&gt; In situ</em&gt; Visualizzazione o la quantificazione di architettura intatta comunicare un sistema.

3-Dimensional Casting resina e imaging di Vena mouse portale o sistema biliare intraepatica Duct

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

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