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

Dette arbejde blev støttet af tilskud fra National Institutes of Health (NIH) til SSH (R01-DK078640), fra Howard Hughes Medical Institute (HHMI) gennem HHMI / Vanderbilt University Certificate Program i molekylær medicin til TJW (GRDOT56006779), den Vanderbilt Diabetes Research and Training Center (P30-DK020593) og Vanderbilt Digestive Disease Research Center (P30-DK058404) at yde Core Services.

1. Forbered Kanyle <ol><li> Varm en 1-tommer lang sektion af PE10 slange med fingerspidserne og strække det, så slangen bliver tynd. Bemærk: størrelsen af karret eller kanalen, der skal kanyle, vil bestemme graden af strækning kræves. Kanylen skal være godt strakt til galdegang kaster, men kan kun være forpligtet til at være moderat strakt til at passe i større portal vene. </li><li> Skær det strakte rør i en diagonal til at generere en affaset spids. </li><li> Skær den anden kant af slang...

<ol>
	<li>Sparks, E. E., Perrien, D. S., Huppert, K. A., Peterson, T. E., Huppert, S. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Defects+in+hepatic+Notch+signaling+result+in+disruption+of+the+communicating+intrahepatic+bile+duct+network+in+mice.">Defects in hepatic Notch signaling result in disruption of the communicating intrahepatic bile duct network in mice.</a> Dis. Model Mech. 4, 359-367 (2011).</li><li>Vanderpool, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+interactions+between+hepatocyte+nuclear+factor-6+and+notch+signaling+regulate+mouse+intrahepatic+bile+duct+development+in+vivo.">Genetic interactions between hepatocyte nuclear factor-6 and notch signaling regulate mouse intrahepatic bile duct development in vivo.</a> Hepatology. 55, 233-243 (2012).</li><li>Meyer, E. P., Ulmann-Schuler, A., Staufenbiel, M., Krucker, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Altered+morphology+and+3D+architecture+of+brain+vasculature+in+a+mouse+model+for+Alzheimer's+disease.">Altered morphology and 3D architecture of brain vasculature in a mouse model for Alzheimer's disease.</a> Proceedings of the National Academy of Sciences. 105, 3587-3592 (2008).</li><li>Van Steenkiste, C. <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:+analyzing+wall+shear+stress+in+the+portal+vein+and+vascular+abnormalities+in+portal+hypertensive+and+cirrhotic+rodents.">Vascular corrosion casting: analyzing wall shear stress in the portal vein and vascular abnormalities in portal hypertensive and cirrhotic rodents.</a> Lab Invest. 90, 1558-1572 (2010).</li><li>Dickie, R., Cormack, M., Semmler-Behnke, M., Kreyling, W. G., Tsuda, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deep+pulmonary+lymphatics+in+immature+lungs.">Deep pulmonary lymphatics in immature lungs.</a> Journal of Applied Physiology. 107, 859-863 (2009).</li><li>Kelly, D. M., McEntee, G. P., McGeenery, K. F., Fitzpatrick, J. M. <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+pancreas,+liver,+and+kidney+in+cerulein-induced+pancreatitis.">Microvasculature of the pancreas, liver, and kidney in cerulein-induced pancreatitis.</a> Archives of Surgery. 128, 293-295 (1993).</li><li>Sparks, E. E., Huppert, K. A., Brown, M. A., Washington, M. K., Huppert, S. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Notch+signaling+regulates+formation+of+the+three-dimensional+architecture+of+intrahepatic+bile+ducts+in+mice.">Notch signaling regulates formation of the three-dimensional architecture of intrahepatic bile ducts in mice.</a> Hepatology. 51, 1391-1400 (2010).</li><li>Carlson, T. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endothelial+expression+of+constitutively+active+Notch4+elicits+reversible+arteriovenous+malformations+in+adult+mice.">Endothelial expression of constitutively active Notch4 elicits reversible arteriovenous malformations in adult mice.</a> Proc. Natl. Acad. Sci. U.S.A. 102, 9884-9889 (2005).</li><li>Hemmeryckx, B., Emmerechts, J., Bovill, E. G., Hoylaerts, M. F., Lijnen, H. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+ageing+on+the+murine+venous+circulation.">Effect of ageing on the murine venous circulation.</a> Histochem. Cell Biol. , (2012).</li><li>Hossler, F. E., Olson, K. R. <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+nasal+salt+gland+of+the+duckling,+Anas+platyrhynchos:+quantitative+responses+to+osmotic+adaptation+and+deadaptation+studied+with+vascular+corrosion+casting.">Microvasculature of the nasal salt gland of the duckling, Anas platyrhynchos: quantitative responses to osmotic adaptation and deadaptation studied with vascular corrosion casting.</a> J. Exp. Zool. 254, 237-247 (1990).</li><li>Hossler, F. E., West, R. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Venous+valve+anatomy+and+morphometry:+studies+on+the+duckling+using+vascular+corrosion+casting.">Venous valve anatomy and morphometry: studies on the duckling using vascular corrosion casting.</a> Am. J. Anat. 181, 425-432 (1988).</li><li>Hossler, F. E., Monson, F. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Structure+and+blood+supply+of+intrinsic+lymph+nodes+in+the+wall+of+the+rabbit+urinary+bladder--studies+with+light+microscopy,+electron+microscopy,+and+vascular+corrosion+casting.">Structure and blood supply of intrinsic lymph nodes in the wall of the rabbit urinary bladder--studies with light microscopy, electron microscopy, and vascular corrosion casting.</a> Anat. Rec. 252, 477-484 (1998).</li><li>Hossler, F. E., Monson, F. 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. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extraction+of+morphometry+and+branching+angles+of+porcine+coronary+arterial+tree+from+CT+images.">Extraction of morphometry and branching angles of porcine coronary arterial tree from CT images.</a> Am J Physiol. Heart. Circ. Physiol. 297, H1949-H1955 (2009).</li><li>Hossler, F. E., Douglas, J. E., Douglas, L. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anatomy+and+morphometry+of+myocardial+capillaries+studied+with+vascular+corrosion+casting+and+scanning+electron+microscopy:+a+method+for+rat+heart.">Anatomy and morphometry of myocardial capillaries studied with vascular corrosion casting and scanning electron microscopy: a method for rat heart.</a> Scan Electron Microsc. , 1469-1475 (1986).</li><li>Hossler, F. E., Douglas, J. 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. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+Assessment+of+the+Rat+Intrahepatic+Biliary+System+by+Three-Dimensional+Reconstruction.">Quantitative Assessment of the Rat Intrahepatic Biliary System by Three-Dimensional Reconstruction.</a> The American Journal of Pathology. 158, 2079-2088 (2001).</li><li>Masyuk, T. V., Ritman, E. L., LaRusso, N. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hepatic+Artery+and+Portal+Vein+Remodeling+in+Rat+Liver:+Vascular+Response+to+Selective+Cholangiocyte+Proliferation.">Hepatic Artery and Portal Vein Remodeling in Rat Liver: Vascular Response to Selective Cholangiocyte Proliferation.</a> The American Journal of Pathology. 162, 1175-1182 (2003).</li></ol>

Vi har beskrevet specifikke eksempler på, hvordan portvenen og intrahepatiske galdegangsceller systemer i leveren kan støbes, men denne teknik kan anvendes på praktisk talt alle andre ductal eller vaskulære system med små justeringer. Tidligere arbejde har påvist gennemførligheden af denne teknik i flere arter, herunder mus 1,2,7-9, ælling 10,11, kanin 12,13, hund 14 og svin 15, og i mange organer, herunder nasal kirtel 14, hjertet 16, ...

<table border="1"><tbody><tr><td> Navn på reagens </td><td> Firma </td><td> Katalog nummer </td><td> Kommentarer </td></tr><tr><td> PE10 polyethylenslange </td><td> Fisher Scientific </td><td> 1417012P </td><td></td></tr><tr><td> 5-0 kirurgisk sort flettet silke </td><td> Roboz Kirurgisk </td><td> SUT-15-1 </td><td></td></tr><tr><td> Steriject 32 G x 13 mm nål </td><td> Air-Tite </td><td> TSK3213 </td><td></td></tr><tr><td> Spring saks </td><td> Fin Science Tools </td><td> 15.000-08 </td><td></td></tr><tr><td> Nummer 5 pincet </td><td> Fin Science Tools </td><td> 91.150-20 </td><td></td></tr><tr><td> MercOx II kit </td><td> Ladd Research </td><td> 21.247 </td><td></td></tr><tr><td> Benzylalkohol </td><td> Fisher Scientific </td><td> 1816-04 </td><td> Only kræves til Babb-clearing </td></tr><tr><td> Benzylbenzoat </td><td> MP Biomedicals, LLC </td><td> 154.839 </td><td> Kun nødvendig for Babb-clearing </td></tr><tr><td> Phosphatpufret saltvand (PBS) </td><td></td><td></td><td></td></tr><tr><td> Modeling ler </td><td></td><td></td><td></td></tr><tr><td> Scale </td><td></td><td></td><td></td></tr><tr><td> Laboratory saks </td><td></td><td></td><td></td></tr><tr><td> 15 ml cap polypropylenrør </td><td></td><td></td><td></td></tr><tr><td> 4% paraformaldahyde </td><td></td><td></td><td></td></tr><tr><td> 15% kaliumhydroxid (KOH) </td><td></td><td></td><td> Kun nødvendig for maceration </td></tr><tr><td> Barberblad </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 sprøjte </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

I organer, er den korrekte arkitektur af vaskulære og ductal strukturer uundværlig for korrekt fysiologisk funktion, og dannelsen og vedligeholdelsen af ​​disse strukturer er en stærkt reguleret proces. Analysen af ​​disse komplekse, tre-dimensionelle strukturer er i høj grad afhængig af enten 2-dimensional behandling i snit eller på farvning injektion undersøgelser. Disse teknikker er imidlertid ikke i stand til at give et komplet og kvantificerbar repræsentation af de duktale eller vaskulære strukturer de skal belyse. Alternativt arten af ​​3-dimensionale plastharpiks afstøbninger frembringer en permanent øjebliksbillede af systemet og er et hidtil ukendt og meget nyttig teknik til visualisering og kvantificering af 3-dimensionelle strukturer og netværk. En afgørende fordel ved harpiksens støbning systemet er evnen til at bestemme den intakte og forbundet, eller kommunikere, strukturen af ​​et blodkar eller kanal. Strukturen af ​​vaskulære og duclæggende net er afgørende for organfunktion, og denne teknik har potentiale til at støtte undersøgelse af vaskulære og ductal net på flere måder. Harpiks støbning kan anvendes til at analysere normal morfologi og funktionelle arkitektur af en luminal struktur, identificere udviklingsmæssige morfogenetiske ændringer og afdække morfologiske forskelle i vævsarkitektur mellem normale og sygdomstilstande. Tidligere arbejde har udnyttet harpiks støbning for at studere, for eksempel, arkitektoniske og funktionelle defekter i mus intrahepatisk galde kanalsystem, som ikke var afspejlet i 2-dimensional analyse af strukturen 1,2, ændringer i hjernen vaskulaturen af en Alzheimers sygdom musemodel 3 , vena abnormiteter i portal hypertensive og cirrotiske mus 4, udviklingsmæssige skridt i rotte lymfatisk modning mellem umodne og voksne lunger 5, umiddelbare mikrovaskulære ændringer i rotte lever, bugspytkirtel og nyre som respons på til kemisk skade6. Her præsenterer vi en metode til at generere en 3-dimensionel harpiks støbt af en mus vaskulær eller ductal netværk, der fokuserer specifikt på portalen vene og intrahepatisk galdegang. Disse afstøbninger kan visualiseres ved at fjerne eller udblødning af væv og kan derefter analyseres. Denne teknik kan anvendes til stort set alle vaskulære eller gangsystemet og ville være direkte anvendelse på enhver undersøgelse spørgende i udvikling, funktion, vedligeholdelse eller skade af en 3-dimensionel ductal eller vaskulær struktur.

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

En fremgangsmåde til visualisering og kvantificering af 3-dimensionale struktur af muse portårevenen eller intrahepatisk galdegang er beskrevet. Denne harpiks støbt teknik kan også anvendes på andre duktale eller vaskulære systemer og muliggør<em&gt; In situ</em&gt; Visualisering eller kvantificering af et systems intakt kommunikere arkitektur.

3-dimensionelt Resin Støbning og Imaging of Mouse Portal Vein eller Intrahepatisk galdegang System

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

Research

JoVE Journal

Medicine

Cannulation of the Mouse Submandibular Salivary Gland via the Wharton's Duct

Severe salivary gland hypofunction is frequently found in patients with Sj&ouml;gren's syndrome and those who receiving therapeutic 
irradiation in their ...

Modeling and Imaging 3-Dimensional Collective Cell Invasion

Modellering og Imaging 3-Dimensional Collective Cell Invasion

A defining characteristic of cancer malignancy is invasion and metastasis 1. In some cancers (e.g. glioma 2), local invasion into surrounding healthy ...

Generation and 3-Dimensional Quantitation of Arterial Lesions in Mice Using Optical Projection Tomography

Generation og 3-Dimensional Kvantificering af Arteriel Læsioner i mus Brug Optisk Projection Tomography

The generation and analysis of vascular lesions in appropriate animal models is a cornerstone of research into cardiovascular disease, generating ...

Heterotopic Auxiliary Rat Liver Transplantation With Flow-regulated Portal Vein Arterialization in Acute Hepatic Failure

Heterotop Auxiliary Rat levertransplanterede patienter med Flow-regulerede portåren Arterialization i akut leversvigt

In acute hepatic failure auxiliary liver transplantation is an interesting alternative approach. The aim is to provide a temporary support until the ...

Bile Duct Ligation in Mice: Induction of Inflammatory Liver Injury and Fibrosis by Obstructive Cholestasis

Bile Duct Ligering i mus: Induktion af inflammatorisk leverskade, og fibrose ved Obstruktiv Kolestase

In most vertebrates, the liver produces bile that is necessary to emulsify absorbed fats and enable the digestion of lipids in the small intestine as well ...

Using Multi-fluorinated Bile Acids and In Vivo Magnetic Resonance Imaging to Measure Bile Acid Transport

Using Multi-fluorinated Bile Acids and In Vivo Magnetic Resonance Imaging to Measure Bile Acid Transport

Brug Multi-fluorerede galdesyrer og<em&gt; In vivo</em&gt; Magnetic Resonance Imaging til måling Bile Acid Transport

Along with their traditional role as detergents that facilitate fat absorption, emerging literature indicates that bile acids are potent signaling ...

Ileectomy-induced Bile Overaccumulation in Mouse Intestine

Ileectomy-induceret galde Overaccumulation i mus tarmen

Intestinal resection is a common therapeutic approach for human diseases such as obesity, inflammatory bowel disease, Crohn's disease, and colon cancer ...

Rat Model of the Associating Liver Partition and Portal Vein Ligation for Staged Hepatectomy (ALPPS) Procedure

Rat Model of the Associating Liver Partition and Portal Vein Ligation for Staged Hepatectomy ALPPS Procedure

Rotte Model af inddrage fiskelever Partition og Portal vene ligatur procedure, iscenesatte Hepatectomy (ALPPS)

Recent clinical data support an aggressive surgical approach to both primary and metastatic liver tumors. For some indications, like colorectal liver ...

Ablation of Ischemic Ventricular Tachycardia Using a Multipolar Catheter and 3-dimensional Mapping System for High-density Electro-anatomical Reconstruction

Ablation af iskæmisk ventrikulær takykardi ved hjælp af en multipolær kateter og 3-dimensionelle Mapping System for High-density Electro-anatomiske genopbygning

Ventricular tachycardia (VT) in patients with ischemic cardiomyopathy mainly results from endocardial scars after myocardial infarction; those scars ...

Laser Doppler Perfusion Imaging in the Mouse Hindlimb

Laser Doppler Perfusion Imaging i musen hindlimb

Blood flow recovery is a critical outcome measure after experimental hindlimb ischemia or ischemia-reperfusion. Laser Doppler perfusion imaging (LDPI) is ...