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

Ringraziamo la sig. ra Sonja Dufentester, Sig. ra Elke Weber e la signora Klaudia Niepagenkämper per l'eccellente assistenza tecnica.

Tutti gli esperimenti sugli animali sono stati approvati dal Landesamt für Natur, Umwelt und Verbraucherschutz (LANUV) Nordrhein-Westfalen secondo la legge di protezione animale tedesco (Tierschutzgesetz).
1. materiali e messa a punto sperimentale
<ol>
	<li>Cura degli animali.
	<ol>
		<li>Utilizzare topi di pari età e del sesso di qualsiasi ceppo DSS-sensibili (ad es., C57BL/6) a 20-25 g di peso corporeo.</li>
		<li>Pianificare almeno cinque o più topi per ogni g...

<ol>
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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>

Anche se le tecniche di imaging medicale si sono evolute rapidamente negli ultimi anni, siamo ancora limitati nella nostra capacità di rilevare processi infiammatori o tumori, nonché altre malattie, nelle loro prime fasi di sviluppo. Tuttavia, questo è fondamentale per la crescita del tumore di comprensione, invasione, o lo sviluppo di metastasi e processi cellulari, nello sviluppo di patologie infiammatorie e malattie degenerative, cardiovascolari e immunologiche. Mentre le tecniche di imaging tradizionale si basano ...

Modelli animali sono ampiamente utilizzati nella ricerca scientifica, ed esistono molte procedure non invasive per monitorare l'attività della malattia e vitalità, come la quantificazione delle variazioni di peso corporeo o l'analisi del sangue, delle urine e le feci. Tuttavia, questi sono solo i parametri di surrogati indiretti che sono anche soggetti a variabilità inter-individuale. Essi frequentemente devono essere accompagnate da analisi post-mortem del campione di tessuto, che impedisce l'osservazione seriale in momenti ripetitivi e diretta osservazione del fisiologico o patologico elabora in vivo. Sofisticate tecniche di imaging di piccoli animali sono emersi, tra cui croce componibile imaging, imaging ottico e l'endoscopia, che consente la visualizzazione diretta di questi processi e consente anche analisi ripetitivi della stessa animali1 , 2 , 3. Inoltre, la possibilità di monitorare in modo ripetitivo vari Stati di malattia nello stesso animale potrebbe diminuire il numero di animali necessari, che potrebbe essere desiderabile da un punto di vista etico degli animali.
Diverse tecniche di imaging ottici differenti esistono per in vivo imaging di fluorescenza. Originariamente, imaging confocale è stata impiegata per studiare la superficie e di sottosuolo eventi fluorescente4,5. Recentemente, tuttavia, tomografiche sistemi che consentono per le valutazioni quantitative del tessuto tridimensionale sono stati sviluppati6. Questo è stato compiuto attraverso lo sviluppo di sonde fluorescenti che emettono luce nello spettro infrarosso vicino (NIR), che offre a basso assorbimento, rivelatori sensibili e sorgenti di luce monocromatica7. Mentre le tecniche di imaging cross-sectioning tradizionali, come la tomografia computata (CT), la formazione immagine a risonanza magnetica (MRI) o ecografia (US), si basano principalmente su parametri fisici e visualizzare la morfologia, imaging ottico può fornire ulteriori informazioni sui processi molecolari sottostanti utilizzando endogene o esogene fluorescenti sonde8.
Progressi della biologia molecolare hanno contribuito a facilitare la generazione di sonde molecolari fluorescenti e mirate per un crescente numero di bersagli. Ad esempio, ricevitore-ha mediato l'assorbimento e la distribuzione in una zona di destinazione specificata può essere visualizzati utilizzando carbocyanine derivato-etichettato anticorpi9. L'abbondanza di anticorpi disponibili, che possono essere etichettati come elementi traccianti specifici in zone altrimenti inaccessibili del corpo, fornisce approfondimenti senza precedenti di processi cellulari e molecolari in modelli di tumorigenesi e neurodegenerative, malattie cardiovascolari, immunologica e infiammatoria7.
In questo studio, descriviamo l'uso di tomografia fluorescenza-mediata in un modello murino di colite. Destrano solfato di sodio (DSS)-colite indotta è un modello standard del mouse chimicamente indotta di infiammazione intestinale simile infiammatorie intestinali (IBD) la malattia10. È particolarmente utile valutare il contributo del sistema immunitario innato allo sviluppo dell'intestino infiammazione11. Poiché il reclutamento, l'attivazione e infiltrazione di monociti e macrofagi rappresentano passaggi cruciali nella patogenesi delle IBD, visualizzazione della loro assunzione e la cinetica di infiltrazione sono essenziali per monitoraggio, ad esempio, l'effetto di sostanze terapeutiche potenziali in un' impostazione preclinici12. Descriviamo l'induzione di colite DSS e dimostrare la caratterizzazione tomografia-mediata di infiltrazione dei macrofagi nella mucosa dell'intestino usando la fluorescenza molecolare tomografia per la visualizzazione di specifica del marcatore del monocito/macrofago F4/80 13. Inoltre, ci illustrano le procedure ausiliari e supplementari, come l'etichettatura dell'anticorpo; la messa a punto sperimentale; e analisi e interpretazione delle immagini ottenute, in correlazione con le letture convenzionali quali gli indici di attività di malattia, flusso cytometry e l'analisi istologica e immunoistochimica. Discutiamo le limitazioni di questa tecnica e i confronti con altre modalità di formazione immagine.

<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

Modelli murini di malattia sono indispensabili alla ricerca scientifica. Tuttavia, molti strumenti diagnostici quali l'endoscopia o formazione immagine tomografica non sono abitualmente impiegati nei modelli animali. Letture sperimentali convenzionali spesso si basano su analisi post-mortem ed ex vivo , che impediscono gli esami di follow-up intra-individuali e aumentano il numero di animali di studio necessari. Fluorescenza-mediata tomografia consente la valutazione non invasiva, ripetitiva, quantitativa e tridimensionale di sonde fluorescenti. È altamente sensibile e consente l'utilizzo di produttori di molecolare, che permette la rilevazione specifica e caratterizzazione di bersagli molecolari distinte. In particolare, sonde mirate rappresentano uno strumento innovativo per l'analisi di espressione genica della proteina e l'attivazione in infiammazione, malattia autoimmune, infezione, malattia vascolare, migrazione cellulare, tumorigenesi, ecc. In questo articolo, forniamo istruzioni dettagliate su questa sofisticata tecnologia di imaging per la rilevazione in vivo e la caratterizzazione di infiammazione (cioè, l'infiltrazione del macrofago di F4/80-positivo) in un modello murino ampiamente usato di infiammazione intestinale. Questa tecnica potrebbe essere utilizzata anche in altre aree di ricerca, quali il rilevamento delle cellule o cellule staminali immune.

Valutazione di colite:
Colite DSS indotta è un modello murino chimicamente indotto di infiammazione intestinale che assomiglia a IBD umana e conduce alla perdita di peso, sanguinamento rettale, ulcerazione superficiale e danno mucoso in topi suscettibili15. È particolarmente utile studiare il contributo del sistema immunitario innato per lo sviluppo di infiammazione intestinale<sup clas...

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

Specifica della destinazione sonde rappresenta uno strumento innovativo per l'analisi dei meccanismi molecolari, come espressione della proteina in vari tipi di malattia (ad es., infiammazione, infezione e tumorigenesi). In questo studio, descriviamo una valutazione tomografica tridimensionale quantitativa di infiltrazione del macrofago intestinale in un modello murino di colite usando la tomografia di fluorescenza-mediata specifica per F4/80.

Fluorescenza-mediata di tomografia per la rilevazione e la quantificazione di infiammazione intestinale murina del macrofago-correlati

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

Quantificazione di attività placca aterosclerotica e l&#39;infiammazione vascolare utilizzando [18-F] Fluorodeoxyglucose Positron Emission Tomography / Computed Tomography (FDG-PET/TC)

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

Ricerca

Medicina

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

Valutazione funzionale della motilità intestinale e Wall infiammazione intestinale nei roditori: Analisi in un modello standardizzato di manipolazione intestinale

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

Luminescenza Imaging Cerenkov (CLI) per il monitoraggio Cancer Therapy

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

Murino Endoscopia per<em&gt; In Vivo</em&gt; Guarigione Imaging Multimodale di Cancerogenesi e valutazione della ferita intestinale e infiammazione

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

Mouse trapianto renale: Modelli di Allograft Rifiuto

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

Un modello murino di irreversibile e reversibile unilaterale Ureteric Ostruzione

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

Modello murino di ischemia-riperfusione intestinale Injury

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

Calcificazione vascolare cellule muscolari lisce e imaging di aortica calcificazioni e infiammazione

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

A Fluorescence-based Assay for Characterization and Quantification of Lipid Droplet Formation in Human Intestinal Organoids

Un saggio basato sulla fluorescenza per la caratterizzazione e la quantificazione della formazione di gocciolini di lipidi negli organiidi intestinali umani

Dietary lipids are taken up as free fatty acids (FAs) by the intestinal epithelium. These FAs are intracellularly converted into triglyceride (TG) ...

A Murine Model of Fetal Exposure to Maternal Inflammation to Study the Effects of Acute Chorioamnionitis on Newborn Intestinal Development

Un modello murino di esposizione fetale all'infiammazione materna per studiare gli effetti della corioamionite acuta sullo sviluppo intestinale appena nato