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 Mrs Klaudia Niepagenkämper for den fremragende teknisk bistand.

Alle dyreforsøg blev godkendt af Landesamt für Natur, Umwelt und Verbraucherschutz (LANUV) Nordrhein-Westfalen ifølge den tyske lov om dyr (Tierschutzgesetz).
1. materialer og eksperimentel opsætning
<ol>
	<li>Dyrs pleje.
	<ol>
		<li>Bruge sex - og alder-matchede mus af enhver DSS-modtagelige stamme (f.eks. C57BL/6) på 20-25 g kropsvægt.</li>
		<li>Plan mindst fem eller flere mus pr. eksperimentelle gruppe og hus mus ifølge lokale dyrs pleje retningslinjer.</...

<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 medicinske Billeddannende teknikker har udviklet sig hurtigt i de seneste år, er vi stadig begrænset vores evne til at detektere inflammatoriske processer eller tumorer, samt andre sygdomme i deres tidligste udviklingsfaser. Dette er imidlertid afgørende at forståelse tumorvækst, invasion, eller udvikling af metastaser og cellulære processer i udviklingen af inflammatoriske sygdomme og degenerative, hjerte-kar- og immunologiske sygdomme. Mens traditionelle Billeddannende teknikker er afhængige af fysiske e...

Dyremodeller er meget udbredt i videnskabelig forskning, og mange ikke-invasive procedurer findes til skærm sygdomsaktivitet og vitalitet, som kvantificering af kroppens vægtændringer eller analyse af blod, urin og afføring. Disse er imidlertid kun indirekte surrogat parametre, der er også omfattet af indbyrdes individuelle variation. De skal ofte suppleres med slagtningen analyser af væv modellen, som forhindrer seriel observation på gentagne gang point og direkte observation af fysiologiske eller patologiske processer in vivo. Sofistikeret lille dyr Billeddannende teknikker er dukket op, herunder grænseoverskridende Sektional imaging, optiske billeddannelse og endoskopi, som giver mulighed for direkte visualisering af disse processer og også giver mulighed for gentagne analyser af de samme dyr1 , 2 , 3. Derudover mulighed for at gentagne overvåge forskellige stater af sygdom i den samme dyr kan reducere antallet af dyr behov, der måtte være ønskeligt fra en Dyreetik synspunkt.
Flere forskellige optiske billeddannelse teknikker findes i vivo fluorescens billeddannelse. Oprindeligt, Konfokal imaging var ansat til at studere overflade og undergrund fluorescerende begivenheder4,5. For nylig har tomografisk systemer, der giver mulighed for kvantitative tredimensionale væv vurderinger blevet udviklet6. Dette er opnået gennem udvikling af fluorescerende sonder, der udsender lys i nær-infrarødt (NIR) spektrum, tilbyder lav absorption, følsomme detektorer og monokromatiske lys kilder7. Mens traditionelle tværsnittet Billeddannende teknikker, såsom computertomografi (CT), magnetisk resonans imaging (MR) eller ultralyd (USA), for det meste er afhængige af fysiske parametre og visualisere morfologi, kan optiske billeddannelse give yderligere oplysninger på underliggende molekylære processer sonder ved hjælp af endogen eller eksogen fluorescerende8.
Fremskridt inden for Molekylærbiologi har bidraget til at lette generation af smart og målrettede fluorescerende molekylære sonder til et stigende antal mål. For eksempel, kan receptor-medieret udbredelse og distribution i en given målområdet visualiseres ved hjælp af carbocyanine derivat-mærket antistoffer9. Overfloden af tilgængelige antistoffer, som kan mærkes for at fungere som specifikke røbestoffer i ellers utilgængelige områder af kroppen, giver hidtil uset indsigt i molekylære og cellulære processer i modeller af tumordannelse og neurodegenerative, hjerte-kar-, immunologiske og inflammatoriske sygdomme7.
I denne undersøgelse beskriver vi brugen af fluorescens-medieret tomografi i en murine model af colitis. Dextran natriumsulfat (DSS)-induceret colitis er en standard kemisk inducerede musemodel af tarmbetændelse, der ligner inflammatoriske tarm sygdom (IBD)10. Det er især nyttigt at vurdere bidrag af den medfødte immunsystem til udviklingen af gut betændelse11. Da ansættelse, aktivering og infiltration af monocytter og makrofager repræsenterer afgørende trin i patogenesen af IBD, er visualisering af deres ansættelse og kinetik af infiltration afgørende for overvågning, for eksempel effekten af potentielle terapeutiske stoffer i en prækliniske indstilling12. Vi beskriver induktion af DSS colitis og påvise-tomografi-medieret karakterisering af makrofag perkolatnedsivning i tarm mucosa ved hjælp af fluorescens molekylære tomografi for specifikke visualisering af monocyt/makrofag markør F4/80 13. Derudover vi illustrere hjælpeansatte og supplerende procedurer, såsom antistof mærkning; opsætningen af eksperimenterende; og analyse og fortolkning af de fremkomne billeder, i sammenhæng med konventionelle udlæsninger som sygdom aktivitet indekser, flow flowcytometri og histologiske analyse og Immunhistokemi. Vi diskutere begrænsninger af denne teknik og sammenligninger med andre billeddiagnostiske modaliteter.

<table border="0" cellpadding="0" cellspacing="0" width="1601">
	<tbody>
		<tr height="21">
			<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 af sygdommen er uundværlige for videnskabelig forskning. Men mange diagnostiske værktøjer som endoskopi eller tomografisk imaging er ikke rutinemæssigt ansat i dyremodeller. Konventionelle eksperimentelle udlæsninger afhængige ofte slagtningen og ex vivo analyser, der forhindre intra individuelle opfølgende undersøgelser og øge antallet af undersøgelse dyr behov. Fluorescens-medieret tomografi gør det muligt for en ikke-invasiv, repetitive, kvantitative, tre-dimensionelle vurdering af fluorescerende sonder. Det er yderst følsom og tillader brug af molekylære beslutningstagere, som giver mulighed for specifikke påvisning og karakterisering af forskellige molekylære mål. Især repræsenterer målrettet sonder et innovativt værktøj til at analysere genekspression aktivering og protein i inflammation, autoimmun sygdom, infektion, karsygdom, celle migration, tumordannelse, osv. I denne artikel giver vi trinvise instruktioner på denne avancerede imaging-teknologi til i vivo påvisning og karakterisering af betændelse (dvs. F4/80-positive makrofager infiltration) i en udbredte murine model af tarmbetændelse. Denne teknik kan også anvendes i andre forskningsområder som immun celle eller stamceller tracking.

Vurdering af Colitis:
DSS-induceret colitis er en kemisk inducerede murine model af tarmbetændelse, der ligner menneskets IBD og fører til vægttab, rektal blødning, overfladiske ulceration og slimhinde skader i modtagelige mus15. Det er især nyttigt at studere bidrag af den medfødte immunsystem til udviklingen af tarmbetændelse10,<sup class="xref"...

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

Target-specifikke sonder repræsenterer et innovativt værktøj til at analysere molekylære mekanismer, såsom protein udtryk i forskellige typer af sygdomme (fx inflammation, infektion og tumordannelse). I denne undersøgelse beskriver vi en kvantitativ tredimensionale tomografisk vurdering af intestinal makrofag infiltration i en murine model af colitis bruger F4/80-specifikke fluorescens-medieret tomografi.

Fluorescens-medieret tomografi til påvisning og kvantificering af makrofag-relaterede Murine tarmbetændelse

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

Kvantificering af aterosklerotisk plaque aktivitet og vaskulær inflammation med [18-F] fluorodeoxyglukose Positron Emission Tomography / Computed Tomography (FDG-PET/CT)

Conventional non-invasive imaging modalities of atherosclerosis such as coronary artery calcium (CAC)1 and carotid intimal medial thickness (C-IMT)2 ...

Functional Assessment of Intestinal Motility and Gut Wall Inflammation in Rodents: Analyses in a Standardized Model of Intestinal Manipulation

Functional Assessment of tarmmotilitet og tarmvæggen Inflammation i Gnavere: Analyser i en standardiseret model af Intestinal Manipulation

Inflammation of the gastrointestinal tract is a common reason for a variety of human diseases. Animal research models are critical in investigating the ...

Cerenkov Luminescence Imaging (CLI) for Cancer Therapy Monitoring

Cerenkov Luminescence Imaging CLI for Cancer Therapy Monitoring

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

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

Murin Endoskopi til<em&gt; In Vivo</em&gt; Multimodal Imaging af Carcinogenesis og vurdering af Intestinal sårheling og betændelse

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 nyretransplantation: modeller af allotransplantatafstødning

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

A Murine Model of Irreversible and Reversible Unilateral Ureteric Obstruction

En murin model af Irreversible og reversibel Ensidig ureteric Obstruktion

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

Murine Model of Intestinal Ischemia-reperfusion Injury

Murin model af intestinal iskæmi-reperfusionsskade

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

Calcification of Vascular Smooth Muscle Cells and Imaging of Aortic Calcification and Inflammation

Forkalkning af vaskulære glatte muskelceller og billeddannelse af aorta Forkalkning og Inflammation

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