Name: fluorescence-mediated tomography for the detection and quantification of macrophage-related murine intestinal inflammation
Uploaded: 2017-12-15T12:30:00
Description: Watch this Scientific Journal Video about Fluorescence-mediated Tomography for the Detection and Quantification of Macrophage-related Murine Intestinal Inflammation at JoVE.com

Vi takker Ms. Sonja Dufentester, Ms. Elke Weber og fru Klaudia Niepagenkämper for utmerket teknisk assistanse.

Alle dyreforsøk ble godkjent av Landesamt für Natur, Umwelt und Verbraucherschutz (LANUV) Nordrhein-Westfalen i henhold til tysk dyr beskyttelse loven (Tierschutzgesetz).
1. materialer og eksperimentelle oppsett
<ol>
	<li>Dyr omsorg.
	<ol>
		<li>Bruk kjønn og alder-matchende mus i DSS-følsomme belastning (f.eks C57BL/6) på 20-25 g kroppsvekt.</li>
		<li>Planlegge minst fem eller flere mus per forsøksgruppen og huset musene i henhold til lokale dyr omsorg retni...

<ol>
	<li>Bruckner, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Murine+endoscopy+for+in+vivo+multimodal+imaging+of+carcinogenesis+and+assessment+of+intestinal+wound+healing+and+inflammation.">Murine endoscopy for in vivo multimodal imaging of carcinogenesis and assessment of intestinal wound healing and inflammation.</a> J Vis Exp. (90), (2014).</li><li>Lewis, J. S., Achilefu, S., Garbow, J. R., Laforest, R., Welch, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Small+animal+imaging.+current+technology+and+perspectives+for+oncological+imaging.">Small animal imaging. current technology and perspectives for oncological imaging.</a> Eur J Cancer. 38 (16), 2173-2188 (2002).</li><li>Bettenworth, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Translational+18F-FDG+PET/CT+imaging+to+monitor+lesion+activity+in+intestinal+inflammation.">Translational 18F-FDG PET/CT imaging to monitor lesion activity in intestinal inflammation.</a> J Nucl Med. 54 (5), 748-755 (2013).</li><li>Vowinkel, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Apolipoprotein+A-IV+inhibits+experimental+colitis.">Apolipoprotein A-IV inhibits experimental colitis.</a> J Clin Invest. 114 (2), 260-269 (2004).</li><li>Korlach, J., Schwille, P., Webb, W. W., Feigenson, G. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+lipid+bilayer+phases+by+confocal+microscopy+and+fluorescence+correlation+spectroscopy.">Characterization of lipid bilayer phases by confocal microscopy and fluorescence correlation spectroscopy.</a> Proc Natl Acad Sci USA. 96 (15), 8461-8466 (1999).</li><li>Ntziachristos, V., Tung, C. H., Bremer, C., Weissleder, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fluorescence+molecular+tomography+resolves+protease+activity+in+vivo.">Fluorescence molecular tomography resolves protease activity in vivo.</a> Nat Med. 8 (7), 757-760 (2002).</li><li>Ntziachristos, V., Bremer, C., Weissleder, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fluorescence+imaging+with+near-infrared+light:+new+technological+advances+that+enable+in+vivo+molecular+imaging.">Fluorescence imaging with near-infrared light: new technological advances that enable in vivo molecular imaging.</a> Eur Radiol. 13 (1), 195-208 (2003).</li><li>Ntziachristos, V., Bremer, C., Graves, E. E., Ripoll, J., Weissleder, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+tomographic+imaging+of+near-infrared+fluorescent+probes.">In vivo tomographic imaging of near-infrared fluorescent probes.</a> Mol Imaging. 1 (2), 82-88 (2002).</li><li>Ballou, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tumor+labeling+in+vivo+using+cyanine-conjugated+monoclonal+antibodies.">Tumor labeling in vivo using cyanine-conjugated monoclonal antibodies.</a> Cancer Immunol Immunother. 41 (4), 257-263 (1995).</li><li>Wirtz, S., Neufert, C., Weigmann, B., Neurath, M. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chemically+induced+mouse+models+of+intestinal+inflammation.">Chemically induced mouse models of intestinal inflammation.</a> Nat Protoc. 2 (3), 541-546 (2007).</li><li>Kawada, M., Arihiro, A., Mizoguchi, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Insights+from+advances+in+research+of+chemically+induced+experimental+models+of+human+inflammatory+bowel+disease.">Insights from advances in research of chemically induced experimental models of human inflammatory bowel disease.</a> World J Gastroenterol. 13 (42), 5581-5593 (2007).</li><li>Nowacki, T. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+5A+apolipoprotein+A-I+(apoA-I)+mimetic+peptide+ameliorates+experimental+colitis+by+regulating+monocyte+infiltration.">The 5A apolipoprotein A-I (apoA-I) mimetic peptide ameliorates experimental colitis by regulating monocyte infiltration.</a> Br J Pharmacol. 173 (18), 2780-2792 (2016).</li><li>Hansch, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+imaging+of+experimental+arthritis+with+near-infrared+fluorescence.">In vivo imaging of experimental arthritis with near-infrared fluorescence.</a> Arthritis Rheum. 50 (3), 961-967 (2004).</li><li>Bialkowska, A. B., Ghaleb, A. M., Nandan, M. O., Yang, V. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Improved+Swiss-rolling+Technique+for+Intestinal+Tissue+Preparation+for+Immunohistochemical+and+Immunofluorescent+Analyses.">Improved Swiss-rolling Technique for Intestinal Tissue Preparation for Immunohistochemical and Immunofluorescent Analyses.</a> J Vis Exp. (113), (2016).</li><li>Diaz-Granados, N., Howe, K., Lu, J., McKay, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dextran+sulfate+sodium-induced+colonic+histopathology,+but+not+altered+epithelial+ion+transport,+is+reduced+by+inhibition+of+phosphodiesterase+activity.">Dextran sulfate sodium-induced colonic histopathology, but not altered epithelial ion transport, is reduced by inhibition of phosphodiesterase activity.</a> Am J Pathol. 156 (6), 2169-2177 (2000).</li><li>Kim, J. J., Shajib, M. S., Manocha, M. M., Khan, W. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Investigating+intestinal+inflammation+in+DSS-induced+model+of+IBD.">Investigating intestinal inflammation in DSS-induced model of IBD.</a> J Vis Exp. (60), e3678 (2012).</li><li>Dieleman, L. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chronic+experimental+colitis+induced+by+dextran+sulphate+sodium+(DSS)+is+characterized+by+Th1+and+Th2+cytokines.">Chronic experimental colitis induced by dextran sulphate sodium (DSS) is characterized by Th1 and Th2 cytokines.</a> Clin Exp Immunol. 114 (3), 385-391 (1998).</li><li>Kojouharoff, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neutralization+of+tumour+necrosis+factor+(TNF)+but+not+of+IL-1+reduces+inflammation+in+chronic+dextran+sulphate+sodium-induced+colitis+in+mice.">Neutralization of tumour necrosis factor (TNF) but not of IL-1 reduces inflammation in chronic dextran sulphate sodium-induced colitis in mice.</a> Clin Exp Immunol. 107 (2), 353-358 (1997).</li><li>Sunderkotter, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Subpopulations+of+mouse+blood+monocytes+differ+in+maturation+stage+and+inflammatory+response.">Subpopulations of mouse blood monocytes differ in maturation stage and inflammatory response.</a> J Immunol. 172 (7), 4410-4417 (2004).</li><li>Willmann, J. K., van Bruggen, N., Dinkelborg, L. M., Gambhir, 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=Molecular+imaging+in+drug+development.">Molecular imaging in drug development.</a> Nat Rev Drug Discov. 7 (7), 591-607 (2008).</li><li>Ntziachristos, V., Ripoll, J., Wang, L. V., Weissleder, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Looking+and+listening+to+light:+the+evolution+of+whole-body+photonic+imaging.">Looking and listening to light: the evolution of whole-body photonic imaging.</a> Nat Biotechnol. 23 (3), 313-320 (2005).</li><li>Ntziachristos, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Going+deeper+than+microscopy:+the+optical+imaging+frontier+in+biology.">Going deeper than microscopy: the optical imaging frontier in biology.</a> Nat Methods. 7 (8), 603-614 (2010).</li><li>Stuker, F., Ripoll, J., Rudin, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fluorescence+molecular+tomography:+principles+and+potential+for+pharmaceutical+research.">Fluorescence molecular tomography: principles and potential for pharmaceutical research.</a> Pharmaceutics. 3 (2), 229-274 (2011).</li><li>Beziere, N., Ntziachristos, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optoacoustic+imaging:+an+emerging+modality+for+the+gastrointestinal+tract.">Optoacoustic imaging: an emerging modality for the gastrointestinal tract.</a> Gastroenterology. 141 (6), 1979-1985 (2011).</li><li>Habtezion, A., Nguyen, L. P., Hadeiba, H., Butcher, E. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Leukocyte+Trafficking+to+the+Small+Intestine+and+Colon.">Leukocyte Trafficking to the Small Intestine and Colon.</a> Gastroenterology. 150 (2), 340-354 (2016).</li><li>Ungar, B., Kopylov, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Advances+in+the+development+of+new+biologics+in+inflammatory+bowel+disease.">Advances in the development of new biologics in inflammatory bowel disease.</a> Ann Gastroenterol. 29 (3), 243-248 (2016).</li><li>Sandborn, W. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vedolizumab+as+induction+and+maintenance+therapy+for+Crohn's+disease.">Vedolizumab as induction and maintenance therapy for Crohn's disease.</a> N Engl J Med. 369 (8), 711-721 (2013).</li><li>Vermeire, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Etrolizumab+as+induction+therapy+for+ulcerative+colitis:+a+randomised,+controlled,+phase+2+trial.">Etrolizumab as induction therapy for ulcerative colitis: a randomised, controlled, phase 2 trial.</a> Lancet. 384 (9940), 309-318 (2014).</li><li>Coskun, M., Vermeire, S., Nielsen, O. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Novel+Targeted+Therapies+for+Inflammatory+Bowel+Disease.">Novel Targeted Therapies for Inflammatory Bowel Disease.</a> Trends Pharmacol Sci. , (2016).</li><li>Vermeire, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+mucosal+addressin+cell+adhesion+molecule+antibody+PF-00547,659+in+ulcerative+colitis:+a+randomised+study.">The mucosal addressin cell adhesion molecule antibody PF-00547,659 in ulcerative colitis: a randomised study.</a> Gut. 60 (8), 1068-1075 (2011).</li><li>Terai, T., Nagano, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Small-molecule+fluorophores+and+fluorescent+probes+for+bioimaging.">Small-molecule fluorophores and fluorescent probes for bioimaging.</a> Pflugers Arch. 465 (3), 347-359 (2013).</li><li>Ren, W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dynamic+Measurement+of+Tumor+Vascular+Permeability+and+Perfusion+using+a+Hybrid+System+for+Simultaneous+Magnetic+Resonance+and+Fluorescence+Imaging.">Dynamic Measurement of Tumor Vascular Permeability and Perfusion using a Hybrid System for Simultaneous Magnetic Resonance and Fluorescence Imaging.</a> Mol Imaging Biol. 18 (2), 191-200 (2016).</li><li>Ale, A., Ermolayev, V., Deliolanis, N. C., Ntziachristos, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fluorescence+background+subtraction+technique+for+hybrid+fluorescence+molecular+tomography/x-ray+computed+tomography+imaging+of+a+mouse+model+of+early+stage+lung+cancer.">Fluorescence background subtraction technique for hybrid fluorescence molecular tomography/x-ray computed tomography imaging of a mouse model of early stage lung cancer.</a> J Biomed Opt. 18 (5), 56006 (2013).</li><li>Chames, P., Van Regenmortel, M., Weiss, E., Baty, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Therapeutic+antibodies:+successes,+limitations+and+hopes+for+the+future.">Therapeutic antibodies: successes, limitations and hopes for the future.</a> Br J Pharmacol. 157 (2), 220-233 (2009).</li><li>Faust, A., Hermann, S., Schafers, M., Holtke, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optical+imaging+probes+and+their+potential+contribution+to+radiotracer+development.">Optical imaging probes and their potential contribution to radiotracer development.</a> Nuklearmedizin. 55 (2), 51-62 (2016).</li><li>Mahler, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+susceptibility+of+inbred+mouse+strains+to+dextran+sulfate+sodium-induced+colitis.">Differential susceptibility of inbred mouse strains to dextran sulfate sodium-induced colitis.</a> Am J Physiol. 274 (3 Pt 1), G544-G551 (1998).</li></ol>

Selv om medisinske Bildeteknikker har utviklet seg raskt de siste årene, er vi fremdeles begrenset i vår evne til å oppdage inflammatoriske prosesser eller svulster, samt andre sykdommer, i deres tidligste stadiene av utviklingen. Dette er imidlertid avgjørende for å forstå tumor vekst, invasjon, eller metastaser utvikling og cellulære prosesser i utviklingen av inflammatoriske lidelser og degenerative, hjerte og immunologiske sykdommer. Mens tradisjonelle Bildeteknikker stole på fysisk eller fysiologiske paramet...

Dyr modeller er mye brukt i forskning, og mange ikke-invasiv prosedyrer eksisterer for å overvåke sykdomsaktivitet og vitalitet, som kvantifisering av kropp vekt endringer eller analyse av blod, urin og avføring. Dette er imidlertid bare indirekte surrogat parametere som også inter-individuelle variasjon. De må ofte suppleres av post mortem analyser av vev, som hindrer føljetong observasjon på repeterende tidspunkt og direkte observasjon av fysiologiske eller patologisk prosesser i vivo. Sofistikert liten-dyr Bildeteknikker har dukket opp, inkludert kryss seksjon bildebehandling og optisk tenkelig endoscopy, som kan direkte visualisering av prosessene og gir også mulighet for gjentatte analyser av samme dyr1 , 2 , 3. i tillegg muligheten til å full av gjentagelser overvåke ulike stater av sykdom i samme dyret kan redusere antall dyr nødvendig, som kan være ønskelig fra et dyreetikk synspunkt.
Det finnes flere forskjellige optiske imaging teknikker for i vivo fluorescens bildebehandling. Opprinnelig var AC confocal imaging ansatt å studere overflate og undergrunnen fluorescerende hendelser4,5. Nylig imidlertid har tomographic systemer som tillater kvantitative tredimensjonale vev vurderinger vært utviklet6. Dette er oppnådd gjennom utvikling av fluorescerende sonder som avgir lys i nær-infrarøde (NIR) spektrum, tilbyr lave absorpsjon, følsom detektorer og monokromatisk lyskilder7. Mens tradisjonelle tverrsnittet Bildeteknikker, som beregnet tomografi (CT), magnetisk resonans imaging (MRI) eller ultralyd (US), stole mest på fysisk parameterene og visualisere morfologi, kan optisk tenkelig gi mer informasjon på underliggende molekylære prosesser sonder bruker endogene eller eksogene fluorescerende8.
Fremskritt innen molekylærbiologi har bidratt til å forenkle generasjonen av smart og målrettet fluorescerende molekylær sonder for et økende antall mål. For eksempel kan reseptor-mediert opptak og distribusjon i et gitt mål visualiseres ved hjelp av carbocyanine derivat-merket antistoffer9. Overflod av tilgjengelige antistoffer, som kan være merket skal fungere som bestemte tracers i ellers utilgjengelige områder av kroppen, gir enestående innsikt i molekylære og cellulære prosesser i modeller av tumorigenesis og nevrodegenerative, hjerte, immunologiske og inflammatoriske sykdommer7.
I denne studien beskriver vi bruken av fluorescens-mediert tomografi i murine modell kolitt. Dekstran natrium sulfat (DSS)-indusert kolitt er en standard kjemisk indusert musemodell intestinale betennelsen som ligner inflammatorisk tarm sykdom (IBD)10. Det er spesielt nyttig å vurdere bidrag av det medfødte immunsystemet til utviklingen av gut betennelse11. Siden rekruttering, aktivisering og infiltrasjon av monocytter og makrofager representerer avgjørende skritt i patogenesen av IBD, er visualisering av rekruttering og the kinetics av infiltrasjon avgjørende for overvåking, for eksempel effekten av potensielle terapeutiske stoffer i en prekliniske innstillingen12. Vi beskriver induksjon av DSS kolitt og demonstrere tomografi-mediert karakterisering av macrophage infiltrasjon i tarmen mucosa bruker fluorescens molekylær tomografi for bestemte effekten av monocyte/macrophage merket F4/80 13. i tillegg vi illustrere ekstra og supplerende prosedyrer, for eksempel antistoff merking; Det eksperimentelle oppsettet; og analyse og fortolkning av fått bildene, i sammenheng med konvensjonelle readouts som sykdom aktivitet indekser, flow cytometri og histologiske analyse og immunohistochemistry. Vi diskutere begrensninger av denne teknikken og sammenlignet med andre tenkelig modaliteter.

<table border="0" cellpadding="0" cellspacing="0" width="1601">
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			<td height="21" style="height:21px;width:469px;">Reagents</td>
			<td style="width:423px;"></td>
			<td style="width:320px;"></td>
			<td style="width:389px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Alfalfa-free diet</td>
			<td>Harlan Laboritories, Madison, USA</td>
			<td style="width:320px;">2014</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Bepanthen eye ointment</td>
			<td style="width:423px;">Bayer, Leverkusen, Germany</td>
			<td style="width:320px;">80469764</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Dextran sulphate sodium (DSS)</td>
			<td>TdB Consulatancy, Uppsala, Sweden</td>
			<td>DB001</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:469px;">Eosin</td>
			<td style="width:423px;">Sigma - Aldrich, Deisenhofen, Germany</td>
			<td style="width:320px;">E 4382</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:469px;">Ethylenediaminetetraacetic acid (EDTA)&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
			<td style="width:423px;">Sigma - Aldrich, Deisenhofen, Germany</td>
			<td style="width:320px;">E 9884</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Florene 100V/V</td>
			<td>Abbott, Wiesbaden, Germany</td>
			<td style="width:320px;">B506</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:469px;">Haematoxylin&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
			<td style="width:423px;">Sigma - Aldrich, Deisenhofen, Germany</td>
			<td style="width:320px;">HHS32-1L</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:469px;">O.C.T. Tissue Tek compound&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
			<td style="width:423px;">Sakura, Zoeterwonde, Netherlands</td>
			<td style="width:320px;">4583</td>
			<td>fixative for histological analyses</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:469px;">Phosphate buffered saline, PBS</td>
			<td style="width:423px;">Lonza, Verviers, Belgium</td>
			<td style="width:320px;">4629</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Sodium Chloride 0,9%</td>
			<td>Braun, Melsungen, Germany</td>
			<td style="width:320px;">5/12211095/0411</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Sodium bicarbonate powder</td>
			<td>Sigma Aldrich Deisenhofen, Germany</td>
			<td style="width:320px;">S5761</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Standard diet</td>
			<td>Altromin, Lage, Germany</td>
			<td style="width:320px;">1320</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Tissue-Tek Cryomold</td>
			<td>Sakura, Leiden, Netherlands</td>
			<td style="width:320px;">4566</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Hemoccult (guaiac paper test)</td>
			<td>Beckmann Coulter, Germany</td>
			<td style="width:320px;">3060</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Biotin rat-anti-mouse anti-F4/80 antibody</td>
			<td>Serotec, Oxford, UK</td>
			<td style="width:320px;">MCA497B</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Biotin rat-anti-mouse anti-GR-1&#160;</td>
			<td>BD Pharmingen, Heidelberg Germany</td>
			<td>553125</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Streptavidin-Alexa546</td>
			<td>Molecular Probes, Darmstadt, Germany</td>
			<td style="width:320px;">S-11225</td>
			<td>excitation/emission maximum:&#160; 556/573nm</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Anti-CD11b rat-anti-mouse antibody TC</td>
			<td>Calteg, Burlingame, USA</td>
			<td style="width:320px;">R2b06</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:469px;">Purified anti-mouse F4/80 antibody</td>
			<td>BioLegend, London, UK</td>
			<td style="width:320px;">123102</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">DAPI</td>
			<td>Sigma-Aldrich, Deisenhoffen, Germany</td>
			<td style="width:320px;">D9542</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">FITC-conjugated anti-Ly6C rat-anti-mouse antibody</td>
			<td>BD Pharmingen, Heidelberg, Germany</td>
			<td>553104</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">FACS buffer</td>
			<td>BD Pharmingen, Heidelberg, Germany</td>
			<td style="width:320px;">342003</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Cy7 NHS Ester</td>
			<td>GE Healthcare&#160;Europe, Freiburg, Germany</td>
			<td style="width:320px;">PA17104</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">MPO ELISA</td>
			<td>Immundiagnostik AG, Bensheim, Germany</td>
			<td>K 6631B</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Cy5.5 labeled anti-mouse F4/80 antibody</td>
			<td>BioLegend, London, UK</td>
			<td>123127</td>
			<td>ready to use labelled Antibodies (alternative)</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Anti-Mouse F4/80 Antigen PerCP-Cyanine5.5</td>
			<td>eBioscience, Waltham, USA</td>
			<td>45-4801-80</td>
			<td>ready to use labelled Antibodies (alternative)</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">DMSO (Dimethyl sulfoxide)</td>
			<td>Sigma-Aldrich, Deisenhoffen, Germany</td>
			<td>67-68-5</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:469px;">Isoflurane</td>
			<td>Sigma-Aldrich, Deisenhoffen, Germany</td>
			<td>792632</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Ethanol</td>
			<td>Sigma-Aldrich, Deisenhoffen, Germany</td>
			<td>64-17-5</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Bovine Serum Albumins (BSA)</td>
			<td>Sigma-Aldrich, Deisenhoffen, Germany</td>
			<td>A4612</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Tris Buffered Saline Solution (TBS)</td>
			<td>Sigma-Aldrich, Deisenhoffen, Germany</td>
			<td>SRE0032</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:469px;">Name</td>
			<td style="width:423px;">Company</td>
			<td style="width:320px;">Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Equipment</td>
			<td></td>
			<td style="width:320px;"></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">FACS Calibur Flow Cytometry System</td>
			<td>BD Biosciences GmbH, Heidelberg, Germany</td>
			<td style="width:320px;"></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">FMT 2000 In Vivo Imaging System</td>
			<td>PerkinElmer Inc., Waltham, MA, USA</td>
			<td style="width:320px;">FMT2000</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">True Quant 3.1 Imaging Analysis Software</td>
			<td>PerkinElmer Inc., Waltham, MA, USA</td>
			<td style="width:320px;">included in FMT2000</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Leica DMLB Fluorescent Microscope</td>
			<td>Leica,&#160; 35578 Wetzlar, Germany&#160;</td>
			<td>DMLB</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Bandelin Sonopuls HD 2070</td>
			<td>Bandelin, 12207 Berlin, Germany</td>
			<td style="width:320px;">HD 2070</td>
			<td>ultrasonic homogenizer</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:469px;">Disposable scalpel No 10</td>
			<td>Sigma-Aldrich, Deisenhoffen, Germany</td>
			<td>Z692395-10EA</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:469px;">Metzenbaum scissors 14cm</td>
			<td>Ehrhardt Medizinprodukte GmbH, Geislingen, Germany</td>
			<td>22398330</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">luer lock syringe 5ml</td>
			<td>Sigma-Aldrich, Deisenhoffen, Germany</td>
			<td style="width:320px;">Z248010</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:469px;">syringe needles</td>
			<td>Sigma-Aldrich, Deisenhoffen, Germany</td>
			<td>Z192368&#160;</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Falcon Tube 50ml</td>
			<td>BD Biosciences, Erembodegem, Belgium</td>
			<td style="width:320px;">352070</td>
			<td></td>
		</tr>
	</tbody>
</table>

fluorescence-mediated tomography for the detection and quantification of macrophage-related murine intestinal inflammation

Murine modeller av sykdommen er uunnværlig til vitenskapelig forskning. Men ansatt mange diagnostiske verktøy som endoskopi eller tomographic imaging ikke rutinemessig i dyremodeller. Konvensjonelle eksperimentelle readouts er ofte avhengige av post mortem og ex vivo analyser, som hindrer intra personlige oppfølging undersøkelser og øke antall studie dyr nødvendig. Fluorescens-mediert tomografi kan ikke-invasiv, repeterende, kvantitativ, tredimensjonale vurdering av fluorescerende sonder. Det er svært følsomme og tillater bruk av molekylære beslutningstakere, som gjør at bestemte gjenkjenning og karakterisering av forskjellige molekylære mål. Spesielt representerer målrettet sonder et nyskapende verktøy for å analysere genet aktivisering og protein uttrykk i betennelse, autoimmune sykdommer, infeksjon, vaskulær sykdom, celle migrasjon, tumorigenesis, etc. I denne artikkelen gi vi instruksjoner på denne avansert bildeteknologi for i vivo gjenkjenning og karakterisering av betennelse (dvs. F4/80-positiv macrophage infiltrasjon) i en brukte murine modell av intestinal betennelser. Denne teknikken kan også brukes i forskningsområder som immun celle eller stilk cellen sporing.

Vurdering av kolitt:
DSS-indusert kolitt er en kjemisk indusert murine intestinale betennelsen som ligner menneskelige IBD og fører til vekttap, rektal blødning, overfladiske sår og mucosal skade på følsomme mus15. Det er spesielt nyttig å studere bidrag av det medfødte immunsystemet til utviklingen av intestinale betennelsen10,1...

Watch this Scientific Journal Video about Fluorescence-mediated Tomography for the Detection and Quantification of Macrophage-related Murine Intestinal Inflammation at JoVE.com

Fluorescence-mediated Tomography for the Detection and Quantification of Macrophage-related Murine Intestinal Inflammation

Mål-spesifikke sonder representerer et nyskapende verktøy for å analysere molekylære mekanismer som protein uttrykk i ulike typer sykdommer (f.eks betennelse, infeksjon og tumorigenesis). I denne studien beskriver vi en kvantitativ tredimensjonale tomographic vurdering av intestinal macrophage infiltrasjon i murine modell av kolitt med F4/80-spesifikke fluorescens-mediert tomografi.

Fluorescens-mediert tomografi av kvantifisering av Macrophage-relaterte Murine intestinale betennelsen

Murine models of disease are indispensable to scientific research. However, many diagnostic tools such as endoscopy or tomographic imaging are not ...

Research

JoVE Journal

Medicine

Quantification of Atherosclerotic Plaque Activity and Vascular Inflammation using [18-F] Fluorodeoxyglucose Positron Emission Tomography/Computed Tomography (FDG-PET/CT)

Quantification of Atherosclerotic Plaque Activity and Vascular Inflammation using [18-F] Fluorodeoxyglucose Positron Emission Tomography/Computed Tomography FDG-PET/CT

Kvantifisering av aterosklerotisk plakk aktivitet og Vaskulær Betennelse med [18-F] Fluorodeoxyglucose emisjontomografi / computertomografi (FDG-PET/CT)

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In  molecular imaging, positron emission tomography (PET) and optical imaging (OI)  are two of the most important and thus most widely used modalities1-3. ...

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Mouse models are widely used to study pathogenesis of human diseases and to evaluate diagnostic procedures as well as therapeutic interventions ...

Mouse Kidney Transplantation: Models of Allograft Rejection

Mus nyretransplanterte: Modeller av allograft-avstøting

Rejection of the transplanted kidney in humans is still a major cause of morbidity and mortality. The mouse model of renal transplantation closely ...

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En Muse Modell av Irreversible og Reversibel Ensidig ureter Obstruksjon

Obstruction of the kidney may affect native or transplanted kidneys and results in kidney injury and scarring. Presented here is a model of obstructive ...

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Intestinal ischemia is a life-threatening condition associated with a broad range of clinical conditions including atherosclerosis, thrombosis, ...

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Cardiovascular disease is the leading cause of morbidity and mortality in the world.  Atherosclerotic plaques, consisting of lipid-laden macrophages and ...