Name: ferric chloride-induced thrombosis mouse model on carotid artery and mesentery vessel
Uploaded: 2015-06-29T14:00:00
Description: Watch this Scientific Journal Video about Ferric Chloride-induced Thrombosis Mouse Model on Carotid Artery and Mesentery Vessel at JoVE.com

The authors would like to acknowledge technical support from Joy Yao and Dr. Karen Alt,&#160;as well as funding from the NHMRC and NHF.

All experiments involving animals were approved by the Alfred Medical Research and Education Precinct Animal Ethics Committee (E/1534/2015/B). All surgical manipulations were performed under full anesthesia and the animals did not experience pain at any stage. All experiments described are non-recovery.
1. Preparation
<ol>
	<li>Cut thin bands of filter paper (1 mm x 2 mm).</li>
	<li>Freshly prepare 2 solutions of ferric chloride of 4% (w/v) and 6% (w/v) diluted into deionized water. Prepare ...

<ol>
	<li>Leadley, R. J., Chi, L., Rebello, S. S., Gagnon, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contribution+of+in+vivo+models+of+thrombosis+to+the+discovery+and+development+of+novel+antithrombotic+agents.">Contribution of in vivo models of thrombosis to the discovery and development of novel antithrombotic agents.</a> J Pharmacol Toxicol Methods. 43 (2), 101-116 (2000).</li><li>Johnson, G. J., Griggs, T. R., Badimon, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+utility+of+animal+models+in+the+preclinical+study+of+interventions+to+prevent+human+coronary+artery+restenosis:+analysis+and+recommendations.+On+behalf+of+the+Subcommittee+on+Animal,+Cellular+and+Molecular+Models+of+Thrombosis+and+Haemostasis+of+the+Scientific+and+Standardization+Committee+of+the+International+Society+on+Thrombosis+and+Haemostasis.">The utility of animal models in the preclinical study of interventions to prevent human coronary artery restenosis: analysis and recommendations. On behalf of the Subcommittee on Animal, Cellular and Molecular Models of Thrombosis and Haemostasis of the Scientific and Standardization Committee of the International Society on Thrombosis and Haemostasis.</a> Thromb Haemost. 81 (5), 835-843 (1999).</li><li>Day, S. M., Reeve, J. L., Myers, D. D., Fay, W. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Murine+thrombosis+models.">Murine thrombosis models.</a> Thromb Haemost. 92 (3), 486-494 (2004).</li><li>Sachs, U. J. H., Nieswandt, B. <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+thrombus+formation+in+murine+models.">In vivo thrombus formation in murine models.</a> Circ Res. 100 (7), 979-991 (2007).</li><li>Furie, B., Furie, B. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanisms+of+thrombus+formation.">Mechanisms of thrombus formation.</a> N Engl J Med. 359 (9), 938-949 (2008).</li><li>Pierangeli, S. S., Liu, X. W., Barker, J. H., Anderson, G., Harris, E. 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E., 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=Novel+single-chain+antibody-targeted+microbubbles+for+molecular+ultrasound+imaging+of+thrombosis:+validation+of+a+unique+noninvasive+method+for+rapid+and+sensitive+detection+of+thrombi+and+monitoring+of+success+or+failure+of+thrombolysis+in+mice.">Novel single-chain antibody-targeted microbubbles for molecular ultrasound imaging of thrombosis: validation of a unique noninvasive method for rapid and sensitive detection of thrombi and monitoring of success or failure of thrombolysis in mice.</a> Circulation. 125 (25), 3117-3126 (2012).</li><li>Wang, X., Smith, P. L., Hsu, M. -. Y., Ogletree, M. L., Schumacher, W. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Murine+model+of+ferric+chloride-induced+vena+cava+thrombosis:+evidence+for+effect+of+potato+carboxypeptidase+inhibitor.">Murine model of ferric chloride-induced vena cava thrombosis: evidence for effect of potato carboxypeptidase inhibitor.</a> J Thromb Haemost. 4 (2), 403-410 (2006).</li><li>Karatas, H., Erdener, S. E., 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=Thrombotic+distal+middle+cerebral+artery+occlusion+produced+by+topical+FeCl(3)+application:+a+novel+model+suitable+for+intravital+microscopy+and+thrombolysis+studies.">Thrombotic distal middle cerebral artery occlusion produced by topical FeCl(3) application: a novel model suitable for intravital microscopy and thrombolysis studies.</a> J Cereb Blood Flow Metab. 31 (6), 1452-1460 (2011).</li><li>Jirouskov&#225;, M., Chereshnev, I., V&#228;&#228;n&#228;nen, H., Degen, J. L., Coller, B. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antibody+blockade+or+mutation+of+the+fibrinogen+gamma-chain+C-terminus+is+more+effective+in+inhibiting+murine+arterial+thrombus+formation+than+complete+absence+of+fibrinogen.">Antibody blockade or mutation of the fibrinogen gamma-chain C-terminus is more effective in inhibiting murine arterial thrombus formation than complete absence of fibrinogen.</a> Blood. 103 (6), 1995-2002 (2004).</li><li>Dubois, C., Panicot-Dubois, L., Merrill-Skoloff, G., Furie, B., Furie, B. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glycoprotein+VI-dependent+and+-independent+pathways+of+thrombus+formation+in+vivo.">Glycoprotein VI-dependent and -independent pathways of thrombus formation in vivo.</a> Blood. 107 (10), 3902-3906 (2006).</li><li>Navarrete, A. -. M., Casari, 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=A+murine+model+to+characterize+the+antithrombotic+effect+of+molecules+targeting+human+von+Willebrand+factor.">A murine model to characterize the antithrombotic effect of molecules targeting human von Willebrand factor.</a> Blood. 120 (13), 2723-2732 (2012).</li><li>Rayes, J., Hollestelle, M. 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=Mutation+and+ADAMTS13-dependent+modulation+of+disease+severity+in+a+mouse+model+for+von+Willebrand+disease+type+2B.">Mutation and ADAMTS13-dependent modulation of disease severity in a mouse model for von Willebrand disease type 2B.</a> Blood. 115 (23), 4870-4877 (2010).</li><li>Lamrani, L., Lacout, 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=Hemostatic+disorders+in+a+JAK2V617F-driven+mouse+model+of+myeloproliferative+neoplasm.">Hemostatic disorders in a JAK2V617F-driven mouse model of myeloproliferative neoplasm.</a> Blood. 124 (7), 1136-1145 (2014).</li><li>Zhang, Y., Ye, 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=Increased+platelet+activation+and+thrombosis+in+transgenic+mice+expressing+constitutively+active+P2Y12.">Increased platelet activation and thrombosis in transgenic mice expressing constitutively active P2Y12.</a> J Thromb Haemost. 10 (10), 2149-2157 (2012).</li><li>Sch&#228;fer, K., Konstantinides, 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=Different+mechanisms+of+increased+luminal+stenosis+after+arterial+injury+in+mice+deficient+for+urokinase-+or+tissue-type+plasminogen+activator.">Different mechanisms of increased luminal stenosis after arterial injury in mice deficient for urokinase- or tissue-type plasminogen activator.</a> Circulation. 106 (14), 1847-1852 (2002).</li><li>Wang, X., Palasubramaniam, 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=Towards+effective+and+safe+thrombolysis+and+thromboprophylaxis:+preclinical+testing+of+a+novel+antibody-targeted+recombinant+plasminogen+activator+directed+against+activated+platelets.">Towards effective and safe thrombolysis and thromboprophylaxis: preclinical testing of a novel antibody-targeted recombinant plasminogen activator directed against activated platelets.</a> Circ Res. 114 (7), 1083-1093 (2014).</li><li>Decrem, Y., 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=Ir-CPI,+a+coagulation+contact+phase+inhibitor+from+the+tick+Ixodes+ricinus,+inhibits+thrombus+formation+without+impairing+hemostasis.">Ir-CPI, a coagulation contact phase inhibitor from the tick Ixodes ricinus, inhibits thrombus formation without impairing hemostasis.</a> J Exp Med. 206 (11), 2381-2395 (2009).</li><li>Ma, 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=Desmolaris,+a+novel+factor+XIa+anticoagulant+from+the+salivary+gland+of+the+vampire+bat+(Desmodus+rotundus)+inhibits+inflammation+and+thrombosis+in+vivo.">Desmolaris, a novel factor XIa anticoagulant from the salivary gland of the vampire bat (Desmodus rotundus) inhibits inflammation and thrombosis in vivo.</a> Blood. 122 (25), 4094-4106 (2013).</li><li>Lei, X., 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=Anfibatide,+a+novel+GPIb+complex+antagonist,+inhibits+platelet+adhesion+and+thrombus+formation+in+vitro+and+in+vivo+in+murine+models+of+thrombosis.">Anfibatide, a novel GPIb complex antagonist, inhibits platelet adhesion and thrombus formation in vitro and in vivo in murine models of thrombosis.</a> Thromb Haemost. 111 (2), 279-289 (2014).</li><li>Waisberg, 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=Plasmodium+falciparum+infection+induces+expression+of+a+mosquito+salivary+protein+(Agaphelin)+that+targets+neutrophil+function+and+inhibits+thrombosis+without+impairing+hemostasis.">Plasmodium falciparum infection induces expression of a mosquito salivary protein (Agaphelin) that targets neutrophil function and inhibits thrombosis without impairing hemostasis.</a> PLoS Pathog. 10 (9), e1004338 (2014).</li><li>Owens, A. P., Lu, Y., Whinna, H. C., Gachet, C., Fay, W. P., Mackman, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Towards+a+standardization+of+the+murine+ferric+chloride-induced+carotid+arterial+thrombosis+model.">Towards a standardization of the murine ferric chloride-induced carotid arterial thrombosis model.</a> J Thromb Haemost. 9 (9), 1862-1863 (2011).</li><li>Wang, X., Xu, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+optimized+murine+model+of+ferric+chloride-induced+arterial+thrombosis+for+thrombosis+research.">An optimized murine model of ferric chloride-induced arterial thrombosis for thrombosis research.</a> Thromb Res. 115 (1-2), 95-100 (2005).</li><li>Tseng, M. T., Dozier, A., Haribabu, B., Graham, U. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transendothelial+migration+of+ferric+ion+in+FeCl3+injured+murine+common+carotid+artery.">Transendothelial migration of ferric ion in FeCl3 injured murine common carotid artery.</a> Thromb Res. 118 (2), 275-280 (2006).</li><li>Bonnard, 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=Leukocyte+mimetic+polysaccharide+microparticles+tracked+in+vivo+on+activated+endothelium+and+in+abdominal+aortic+aneurysm.">Leukocyte mimetic polysaccharide microparticles tracked in vivo on activated endothelium and in abdominal aortic aneurysm.</a> Acta Biomater. 10 (8), 3535-3545 (2014).</li><li>Boulaftali, Y., Lamrani, L., 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+mouse+dorsal+skinfold+chamber+as+a+model+for+the+study+of+thrombolysis+by+intravital+microscopy.">The mouse dorsal skinfold chamber as a model for the study of thrombolysis by intravital microscopy.</a> Thromb Haemost. 107 (5), 962-971 (2012).</li><li>Konstantinides, S., Sch&#228;fer, K., Thinnes, T., Loskutoff, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Plasminogen+activator+inhibitor-1+and+its+cofactor+vitronectin+stabilize+arterial+thrombi+after+vascular+injury+in+mice.">Plasminogen activator inhibitor-1 and its cofactor vitronectin stabilize arterial thrombi after vascular injury in mice.</a> Circulation. 103 (4), 576-583 (2001).</li><li>Li, W., McIntyre, T. M., Silverstein, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ferric+chloride-induced+murine+carotid+arterial+injury:+A+model+of+redox+pathology.">Ferric chloride-induced murine carotid arterial injury: A model of redox pathology.</a> Redox Biol. 1 (1), 50-55 (2013).</li></ol>

The ferric chloride induced thrombosis model is an excellent research tool. As shown in this study, it is extremely easy to implement and when used in combination with intravital microscopy or Doppler flowmeter, it provides a good real-time monitoring of thrombus formation. Adjusting the time exposure and the concentration of FeCl3, it also offers the possibility to produce either non-occlusive or occlusive thrombi.
However, this method also has some limitations. In the carotid arte...

The study of the mechanisms involved in the development of thrombosis and the evaluation of the effectiveness of anti-thrombotic drugs requires well established experimental animal models. Large animal models were the first to be used as they provide large vessels more similar to humans than rodents1. However, high cost, the larger facilities required and the difficulty in manipulating them genetically are major drawbacks to their use and large animals are now limited to late preclinical studies once preliminary tests on rodents have given conclusive results2. With wide availability of transgenic and knockout strains and their small size that minimizes the quantity of antithrombotic drugs required for in vivo testing, mice are mainly used for thrombosis research. Therefore, several models of thrombotic disorders have been developed in mice3.
Many established thrombosis models disrupt the intima layer of the vessel wall, followed by the exposure of the sub endothelial extracellular matrix to the blood flow inducing the formation of blood clots4. The thrombi may result from the exposure of collagen which triggers platelets activation or/and from the exposure of tissue factor which activates the coagulation cascade5. Several techniques are then employed to achieve the initial vessel injury. Pierangeli et al. developed a mechanical disruption model with a microsurgery tool on the femoral vein6. Kikushi et al. described a model which consists in the administration of a photo reactive compound (Rose Bengal) which accumulates in the lipid bilayer of endothelial cells followed by the specific excitation of the vessel wall of interest with green light (540 nm)7. The injury can also be induced by a short high-intensity pulse laser illumination8. Another technique firstly established on the carotid artery of rats consists in the topical application of ferric chloride (FeCl3)9. In this case, the vessel denudation results from free radicals generated by FeCl3 which causes lipid peroxidation and destruction of endothelial cells10. The injury induces the expression of several adhesion molecules triggering platelet adhesion and aggregation as well as leukocytes recruitment. It has been demonstrated that leukocytes, particularly neutrophils, play a crucial role in the activation of the blood coagulation cascade leading to thrombosis11. This method is well suited to reproduce the coagulation cascade; investigators must keep in mind that, in this mouse model, thrombosis is typically induced in healthy vessels whereas thrombosis in humans is mainly occurring in diseased e.g. atherosclerotic vessels.
As this model is very simple to implement and is also effective in mice, it is now the mostly used thrombosis model for small animal in vivo studies. In addition, this technique offers the possibility to induce the formation of thrombi in a variety of vessels. Target vessels can be arteries or veins of large diameter (carotid, femoral, vena cava) or small diameter (mesentery, cremaster)12&#8211;14. More recently, it was also used on the proximal middle cerebral artery to develop a model of stroke15. The thrombosis formation may be directly observed by intravital microscopy after fluorescent labeling of platelet and leukocytes or monitored by measuring the blood flow decrease with a temperature probe or a Doppler probe12,16,17. Several parameters such as occlusion time, thrombus formation time or thrombus size may then be investigated. The physiological differences between the vessels investigated result in significant variations in the thrombi obtained. Therefore, investigators usually select the target vessel according to the parameters they want to measure and/or the disease setting they want to investigate. Typically, the model on the carotid artery is more relevant for research on atherothrombosis related to myocardial infarction or stroke whereas studies on the vena cava are more relevant for research on deep venous thrombosis. The accessibility of the different vessels also determines the method used to measure thrombus growth. For instance, the mesenteric vessels are easy to access making this model well suited for intravital microscopic observation and the study of the dynamics of thrombus formation. The carotid artery is less accessible but bigger enabling blood flow measurements and provide an excellent model to study occlusive thrombosis.
The ferric chloride induced thrombosis model has provided tremendous progress in the understanding of this pathology. It has been used in many studies focusing on the role of von Willebrand factor in thrombosis formation18,19. Combined with genetic modification techniques, it has allowed the identification of many specific gene involved in thrombotic disorders. Lamrani et al. for example have shown that a knock-in of the JAK2V617F gene is associated with an accelerating formation of unstable clot20. Zhang et al. have investigated the physiological implication of the P2Y12 platelet receptor and demonstrated that transgenic mice overexpressing specifically this receptor in platelets only, displayed a more rapid and stable thrombus formation in mesenteric artery injured with FeCl321. The crucial role of Tissue-type plasminogen activator (tPA) and urokinase-type plasminogen activator (uPA) in the fibrin degradation process has also been investigated in this method22. Furthermore this model also provides a simple and accurate way of testing the fibrinolytic capacities of many novel drugs in vivo. For instance, Wang et al. have used this model for the preclinical validation of a novel recombinant plasminogen activator targeted against activated platelets23. This method also enabled the validation of therapeutic proteins isolated from the salivary of ticks, vampire bats, and mosquitos or from the venom of snakes with specific identification of the target24-27. These examples demonstrate the versatility of the ferric chloride model. In this article, we focus on two methods and study ferric chloride induced thrombosis on two different vessel type; mesenteric vessel and carotid artery.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Whatman chromatography paper</td>
			<td>GE Healthcare</td>
			<td>3030917</td>
			<td></td>
		</tr>
		<tr>
			<td>Iron (III) chloride 40 % (w/v)</td>
			<td>VWR</td>
			<td>24212.298</td>
			<td></td>
		</tr>
		<tr>
			<td>Rhodamine 6G</td>
			<td>Sigma</td>
			<td>R4127</td>
			<td></td>
		</tr>
		<tr>
			<td>Inverted microscope&#160;</td>
			<td>Olympus</td>
			<td>IX81</td>
			<td></td>
		</tr>
		<tr>
			<td>Digital black-and-white camera&#160;</td>
			<td>Olympus</td>
			<td>XM10</td>
			<td></td>
		</tr>
		<tr>
			<td>Doppler flowmeter</td>
			<td>Transonic</td>
			<td>TS420</td>
			<td></td>
		</tr>
		<tr>
			<td>Nano-doppler flow probe</td>
			<td>Transonic</td>
			<td>0.5 PBS</td>
			<td></td>
		</tr>
		<tr>
			<td>Ketamine</td>
			<td>Hospira&#160;</td>
			<td>0409-2051-05</td>
			<td></td>
		</tr>
		<tr>
			<td>Xylazine (Rampun)</td>
			<td>Bayer</td>
			<td>75313&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>Petri dish</td>
			<td>Sarstedt</td>
			<td>82.1472</td>
			<td></td>
		</tr>
		<tr>
			<td>Insulin syringe (29 G)</td>
			<td>BD Ultra-Fine</td>
			<td>326103</td>
			<td></td>
		</tr>
		<tr>
			<td>Cotton tipped applicators</td>
			<td>BSN medical</td>
			<td>211827A</td>
			<td></td>
		</tr>
		<tr>
			<td>Dynek dysilk sutures</td>
			<td>Dynek Pty Ltd</td>
			<td>CS30100</td>
			<td></td>
		</tr>
		<tr>
			<td>Dulbecco&#39;s phosphate buffer saline (PBS)</td>
			<td>Gibco life technologies</td>
			<td>21600-069</td>
			<td></td>
		</tr>
		<tr>
			<td>Heating pad</td>
			<td>Kirchner</td>
			<td>T60</td>
			<td></td>
		</tr>
	</tbody>
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ferric chloride-induced thrombosis mouse model on carotid artery and mesentery vessel

Severe thrombosis and its ischemic consequences such as myocardial infarction, pulmonary embolism and stroke are major worldwide health issues. The ferric chloride injury is now a well-established technique to rapidly and accurately induce the formation of thrombi in exposed veins or artery of small and large diameter. This model has played a key role in the study of the pathophysiology of thrombosis, in the discovery and validation of novel antithrombotic drugs and in the understanding of the mechanism of action of these new agents. Here, the implementation of this technique on a mesenteric vessel and carotid artery in mice is presented. The method describes how to label circulating leukocytes and platelets with a fluorescent dye and to observe, by intravital microscopy on the exposed mesentery, their accumulation at the injured vessel wall which leads to the formation of a thrombus. On the carotid artery, the occlusion caused by the clot formation is measured by monitoring the blood flow with a Doppler probe.

The fluorescent intravital microscopy observation of the mesentery will reveal the accumulation of Rhodamine 6G labeled platelets and leukocytes along the vessel wall injured by FeCl3. The progressive formation of a partial thrombus is monitored in a 200 &#181;m mesentery vessel (Figure 1). A thrombus slowly appears and is clearly identifiable after the first minute of exposure to FeCl3 (Figure 1, t = 60 sec). 40 sec after the removal of the filter paper soaked with...

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Ferric Chloride-induced Thrombosis Mouse Model on Carotid Artery and Mesentery Vessel

The FeCl3 induced thrombosis model in mice is described herein. A method to monitor thrombus growth by intravital microscopy observation on a mesenteric vessel and by blood flow measurement in the carotid artery is presented.

Severe thrombosis and its ischemic consequences such as myocardial infarction, pulmonary embolism and stroke are major worldwide health issues. The ferric ...

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