Yazarlar trikrom boyama ve analizi ile yardım için Washington Üniversitesi Histoloji Brian Johnson ve Görüntüleme Çekirdek teşekkür ederiz. Bu çalışma NIH hibe HL098067 ve HL089455 tarafından desteklenmiştir.

Washington ve Cedars-Sinai Tıp Merkezi Üniversitesi Kurumsal Hayvan Bakım ve Kullanım Komiteleri (IACUC) bu çalışmalar için gerekli hayvan çalışmaları onayladı. 1. Hazırlık <ol><li> Künt uç forseps ve basacağı yoluyla otoklav hem sterilize. </li><li> biyolojik güvenlik kabini kullanarak, liyofilize toz PBS içinde BLM bir çalışma stok hazırlamak. Karıştırma daha emin olmak için 35 kHz&#39;de 10 dakika süre ile çözelti sonikasyon. Not: ul 30 ile 45 toplam hacmi oldukça büyük hacimli pipet düşük ucunda varyasyon ve boğulma önlemek için tavsiye edilir. </li><li> Yaklaşık 1 prosedürü kendisi için m 2, yanı sıra işlem öncesi ve sonrası her iki kafesleri için belirlenen yerleri içeren temiz bir çalışma alanı hazırlayın. </li><li> taban ve Underl genelinde laboratuvar bant 2 ya da 3 şerit koyarak hemen araştırmacı önünde tezgah prosedür kurulu tabanını Fixtezgah ying. Bir tahta oluşturma konusunda daha fazla özellikleri için Bkz: Şekil 1. </li><li> prosedür kurulu iki konumlandırma vida arasındaki büyüklük 4.0 dikiş ipliğinin tek uzunluğunu kravat. </li><li> kaldırılması ve üç 1 ml&#39;lik şırınga pistonu atarak ve hava geçirmez bir mühür formu her varil üst içine PBS 60 ul yatırarak derme çatma bir spirometre oluşturun. gevşek şırınga birine kateter göbeğini Güvenli ve yönetim kurulu bir tarafa koyun. </li><li> 1 ml şırınganın içine havanın aspire 300 ul ve yönetim kurulu bir tarafa koyun. </li><li> bir tarafa bant uzunluğu ve yerinde yaklaşık 6 inç ek bir parça kesin. Bu aşamada 2.4 kuruluna hayvan sabitlemek için kullanılır. </li><li> Bir izofluran odasını kurmak. Pozlama odasına ve boşluk vakum hem de uygun portlara O 2, izofluran ve vakum takın. Alternatif olarak, anestezik yönetmek bir izofluran uyumlu biyolojikgüvenlik kabini. </li></ol> 2. Entübasyon <ol><li> o bilincini kaybeder ve solunum uygun bir orana yavaşlatır kadar odasında izofluran ile fare anestezisi. Tipik bir pozlama 3-4 dakika süreyle% 4 izofluran ve% 2 O 2 içerir ve ideal bir sonuç sedasyon 2 ila 2,5 dk. Bu 1 nefes her 2 sn bir solunum oranına karşı gelmektedir. </li><li> bir tarafa bir pipet ve yerine BLM 30 ve 45 ul arasında ayarlamak için sedasyon, aspirat için beklerken. </li><li> Hazır olduğunda, işlem platformu konumlandırma vida bağlı akıştan üst kesici dişlerin tarafından sedasyon fare askıya. Hayvanın sırtı platformu yüzeye karşı düz durduğundan emin olun. </li><li> havalandırma kısıtlamak için dikkatli olmak, sadece diyafram üstünde, gevşek göğüs boşluğunun alt (kaudal) kısmı boyunca bir parça bant yerleştirin. Yerleşim proc sırasında düzgün hizalama korumak için sıkı yeterli olmalıdıredure, ancak solunumu kısıtlar o kadar sıkı değil. </li><li> 80 ile% ve% 100 yoğunluğuna aydınlatıcı açın ve solar plexus yakın cilt yüzeyinden 1 2 cm, böylece deveboynu yönlendirmek. Periyodik olarak fare zarar vermez emin olmak için ısı deveboynu ucu kontrol edin. </li><li> platformun arkasında duran, dil bulmak için steril, künt uç forseps kullanabilir. Alt kesici dişler, yavaşça kavrama önlemek ve ağız boşluğunun dışında dilini çekmek için dikkatli olmak. </li><li> Kalan elinizi kullanarak, basacağı yerleştirin ve ağız boşluğunun zemin karşı dilini düzleştirmek için kullanabilirsiniz. forseps bırakın, ancak sonraki iki aşama için yerinde basacağı bırakın. </li><li> trakea o Ana kök bronşların seviyesine ulaşıncaya kadar güneş pleksus seviyesinden proksimal deveboynu rehberlik ederek görünür böylece ışığı yönlendirmek. Not: trakea kolayca solunum aksiyonu ile ayırt edilebilir, WHIch şiddeti dalgalanmaya yayılan ışığı neden olur. Doğru yerleştirilmiş, bu yapının ağız boşluğunda kendisi minimal ortam ışığı ile ışık merkezi bir pim gibi eksenel düzlemde ayırt olacaktır. </li><li> Açı düz lümenine, damlacık içeren ekli şırınga ile, trakea doğal yolu izler ve 22-G kateter ucu düşürmek, böylece şırınga. PBS kabarcık yükselişi ve başarılı yerleştirme üzerine her nefeste düşmeye başlayacaktır. Not: Bu eylem, derin sedasyon sonucunda birkaç saniye kadar gecikebilir. </li><li> ek 5 mm kateter besleyin. dil çıkarın depressor. </li><li> yavaşça şırınga kaldırmak, ters el şırınga Shift ve göbek sürükleyici. </li><li> Kateter hub iç merkezi haline BLM 30 ve 45 ul arasındaki mevduat, ikinci şırınga takın ve göbek içine hava 300 ul dağıtmak. </li><li> ikinci değiştirinİlk PBS balonu içeren şırınga. kabarcık yükselişi ve işlemi başarıyla gerçekleştirildi olup olmadığını düşmeye devam edecektir. </li></ol> 3. Post-prosedürel Bakımı <ol><li> kateter ve bandı çıkarın ve bilinci yerine gelene kadar bir kuru ılık bir yerde hayvan yerleştirin - tipik birkaç dakika içinde. </li></ol>

<ol>
	<li>Hyde, D. M., Hamid, Q., Irvin, C. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anatomy,+pathology,+and+physiology+of+the+tracheobronchial+tree:+emphasis+on+the+distal+airways.">Anatomy, pathology, and physiology of the tracheobronchial tree: emphasis on the distal airways.</a> J Allergy Clin Immunol. 124, S72-S77 (2009).</li><li>Rosenthal, N., Brown, S. <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+ascending:+perspectives+for+human-disease+models.">The mouse ascending: perspectives for human-disease models.</a> Nat Cell Biol. 9, 993-999 (2007).</li><li>Peters, L. 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+as+a+model+for+human+biology:+a+resource+guide+for+complex+trait+analysis.">The mouse as a model for human biology: a resource guide for complex trait analysis.</a> Nat Rev Genet. 8, 58-69 (2007).</li><li>Baron, R. M., Choi, A. J., Owen, C. A., Choi, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetically+manipulated+mouse+models+of+lung+disease:+potential+and+pitfalls.">Genetically manipulated mouse models of lung disease: potential and pitfalls.</a> Am J Physiol Lung Cell Mol Physiol. 302, 485-497 (2012).</li><li>Sharma, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+respiratory+distress+syndrome.">Acute respiratory distress syndrome.</a> BMJ Clin Evid. 2010, (2010).</li><li>Saguil, A., Fargo, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+respiratory+distress+syndrome:+diagnosis+and+management.">Acute respiratory distress syndrome: diagnosis and management.</a> Am Fam Physician. 85, 352-358 (2012).</li><li>Wilson, M. S., Wynn, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pulmonary+fibrosis:+pathogenesis,+etiology+and+regulation.">Pulmonary fibrosis: pathogenesis, etiology and regulation.</a> Mucosal Immunol. 2, 103-121 (2009).</li><li>Wuyts, W. 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=The+pathogenesis+of+pulmonary+fibrosis:+a+moving+target.">The pathogenesis of pulmonary fibrosis: a moving target.</a> Eur Respir J. 41, 1207-1218 (2013).</li><li>Mouratis, M. A., Aidinis, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modeling+pulmonary+fibrosis+with+bleomycin.">Modeling pulmonary fibrosis with bleomycin.</a> Curr Opin Pulm Med. 17, 355-361 (2011).</li><li>Moeller, A., Ask, K., Warburton, D., Gauldie, J., Kolb, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+bleomycin+animal+model:+a+useful+tool+to+investigate+treatment+options+for+idiopathic+pulmonary+fibrosis?.">The bleomycin animal model: a useful tool to investigate treatment options for idiopathic pulmonary fibrosis?.</a> Int J Biochem Cell Biol. 40, 362-382 (2008).</li><li>Myll&#228;rniemi, M., Kaarteenaho, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pharmacological+treatment+of+idiopathic+pulmonary+fibrosis+-+preclinical+and+clinical+studies+of+pirfenidone,+nintedanib,+and+N-acetylcysteine.">Pharmacological treatment of idiopathic pulmonary fibrosis - preclinical and clinical studies of pirfenidone, nintedanib, and N-acetylcysteine.</a> European Clinical Respiratory Journal. 2, (2015).</li><li>Reinert, T., Baldotto, C. S. d. R., Nunes, F. A. P., Scheliga, A. A. d. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bleomycin-Induced+Lung+Injury.">Bleomycin-Induced Lung Injury.</a> Journal of Cancer Research. 2013, 1-9 (2013).</li><li>Lakatos, H. F., 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=Oropharyngeal+aspiration+of+a+silica+suspension+produces+a+superior+model+of+silicosis+in+the+mouse+when+compared+to+intratracheal+instillation.">Oropharyngeal aspiration of a silica suspension produces a superior model of silicosis in the mouse when compared to intratracheal instillation.</a> Exp Lung Res. 32, 181-199 (2006).</li><li>Helms, M. N., Torres-Gonzalez, E., Goodson, P., Rojas, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+tracheal+instillation+of+solutes+into+mouse+lung.">Direct tracheal instillation of solutes into mouse lung.</a> J Vis Exp. , (2010).</li><li>Osier, M., Oberdorster, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intratracheal+inhalation+vs+intratracheal+instillation:+differences+in+particle+effects.">Intratracheal inhalation vs intratracheal instillation: differences in particle effects.</a> Fundam Appl Toxicol. 40, 220-227 (1997).</li><li>Driscoll, K. 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=Intratracheal+instillation+as+an+exposure+technique+for+the+evaluation+of+respiratory+tract+toxicity:+Uses+and+Limitations.">Intratracheal instillation as an exposure technique for the evaluation of respiratory tract toxicity: Uses and Limitations.</a> Toxicol Sci. 55, 24-35 (2000).</li><li>MacDonald, K. D., Chang, H. Y., Mitzner, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+improved+simple+method+of+mouse+lung+intubation.">An improved simple method of mouse lung intubation.</a> J Appl Physiol (1985). 106, 984-987 (1985).</li><li>Spoelstra, E. N., 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+novel+and+simple+method+for+endotracheal+intubation+of+mice.">A novel and simple method for endotracheal intubation of mice.</a> Lab Anim. 41, 128-135 (2007).</li><li>Cai, Y., Kimura, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Noninvasive+intratracheal+intubation+to+study+the+pathology+and+physiology+of+mouse+lung.">Noninvasive intratracheal intubation to study the pathology and physiology of mouse lung.</a> J Vis Exp. , e50601 (2013).</li><li>Starcher, B., WIlliams, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+method+for+intratracheal+instillation+of+endotoxin+into+the+lungs+of+mice.">A method for intratracheal instillation of endotoxin into the lungs of mice.</a> Lab Anim. 23, 234-240 (1989).</li><li>Daubeuf, F., Frossard, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Performing+bronchoalveolar+lavage+in+the+mouse.">Performing bronchoalveolar lavage in the mouse.</a> Curr Protoc Mouse Biol. 2, 167-175 (2012).</li><li>Li, 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=Severe+lung+fibrosis+requires+an+invasive+fibroblast+phenotype+regulated+by+hyaluronan+and+CD44.">Severe lung fibrosis requires an invasive fibroblast phenotype regulated by hyaluronan and CD44.</a> J Exp Med. 208, 1459-1471 (2011).</li><li>Grazioli, 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=CYR61+(CCN1)+overexpression+induces+lung+injury+in+mice.">CYR61 (CCN1) overexpression induces lung injury in mice.</a> Am J Physiol Lung Cell Mol Physiol. 308, L759-L765 (2015).</li><li>Redente, E. F., 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+necrosis+factor-alpha+accelerates+the+resolution+of+established+pulmonary+fibrosis+in+mice+by+targeting+profibrotic+lung+macrophages.">Tumor necrosis factor-alpha accelerates the resolution of established pulmonary fibrosis in mice by targeting profibrotic lung macrophages.</a> Am J Respir Cell Mol Biol. 50, 825-837 (2014).</li><li>Lawrenz, M. B., Fodah, R. A., Gutierrez, M. G., Warawa, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intubation-mediated+intratracheal+(IMIT)+instillation:+a+noninvasive,+lung-specific+delivery+system.">Intubation-mediated intratracheal (IMIT) instillation: a noninvasive, lung-specific delivery system.</a> J Vis Exp. , e52261 (2014).</li></ol>

The authors have nothing to disclose.

Aerosol haline getirme sınırlı olması nedeniyle reaktif temin edilebilirliği, emniyeti ve maliyet için pratik örneklerde, doğrudan nefes borusu idaresi akciğerlere dışsal ajanların uygulanması için üstün bir yöntemdir 16 Transtrakeal instilasyon yaygın Bunu gerçekleştirmek için kullanılmaktadır.; Ancak, tüm cerrahi müdahale ile olduğu gibi, o da onunla prosedürün kendisinden kaynaklanan komplikasyonlar için potansiyel taşır ve mutlaka ajan telkin ediliyor. 13 ...

Bazı anatomik ve fizyolojik farklılıklar olmasına rağmen, 1 fare modelleri hayvancılık açısından bakıldığında, insan biyolojisi ve hastalık patogenezi. 2 modelleme için çok değerli, fareler, kullanımı kolay olan, düşük üreme zamanı, hızlandırılmış ömrü var ve nispeten ucuzdur devam eve. Farklı genetik suşları ve stratejiler (örn., Koşullu knock-out raportör fareler, soy izleme yaklaşımlar, vs.), ve bunun yanı sıra reaktifler geniş (örn., Antikorlar, rekombinant proteinler, inhibitörlerin geliştirilmesi ile, vb), fareler, insan homeostasis ve hastalık süreçlerini ortaya çıkarmak için önemli bir örnek omurgalı organizma haline gelmiştir. 3 Fareler insanlarda 4 ALİ travma, yaralanma veya sepsis neden olabilir. Akut akciğer hasarı (ALI) ve pulmoner fibrozis dahil olmak üzere akciğer koşulları, çalışmak için özellikle değerli olmuştur ve epitel ile karakterizedir veendotel sızıntı (yani., ödem), enflamasyon ve oluşmakta olan fibroz. Birçok hastada, ALI genellikle solunum yetmezliği nedeni ile fibroz ve ölümle sonuçlanan onun ağır formu, akut solunum sıkıntısı sendromu (ARDS), ilerler. 5,6 pulmoner fibrozis ekstraselüler matriksin aşırı birikmesi ile karakterize ilerleyici, ölümcül patolojidir en önemlisi. Ben engelli akciğer fonksiyonu yol açan kolajen bleomisin (BLM) 7,8 Yönetim yazın deney hayvanlarında ALI ve fibroz uyarmak için en yaygın kullanılan ve en iyi karakterize modeldir. 9 kemirgenlerde pulmoner fibrozis BLM kaynaklı olsa da yapar tamamen insan fibrotik fenotipleri, bu model ile 10 fare çalışmaları başlangıcı ve hastalığın ilerlemesini etkileyen birçok önemli faktörlerden keşfine yol açmıştır özetlemek değil. 11 BLM kaynaklı fibrogenezisin arkasında kesin mekanizması (ler) bilinmemekle birlikte, başlangıç ​​hasarınıniletken hava yolları ve alveolleri ve özellikle de tip 1 pnömositler döşeyen epitel hücrelerinde kontak bağımlı DNA zincir kırıkları kaynaklandığı düşünülmektedir. 12 BLM ve pulmoner epitel arasında doğrudan temas ihtiyacı sağlam bir dağıtım yolunun önemini vurgulamaktadır ve bu endişeleri de rekombinant proteinlerin, antikorların, siRNA, virüs, bakteri, partikül, ve daha fazlasını içeren distal hava hedeflenen tedavilerin geniş bir yelpazede, germane vardır. Orofaringeal aspirasyon (OPA), yaygın olarak, bu amaçla 13 kullanılmaktadır, ancak OPA önemli bir eksiklik teslim maddenin bir bölümü bu şekilde uygulanan dozun belirginsizlik yol mide bağırsak Yutulan edilebilir olmasıdır. Yaygın olarak kullanılan bir yaklaşım, doğrudan solunum yollarına bir ajanın trakea ve damlatma maruz güçlü anestezi altında trakeostomi gerektirir transtrakeal damlatma vardır. 14 Ancak, sadece bu tür olabilirBir prosedür nedeniyle invaziv olması istenmeyen olabilir, ama o da zaman tüketen bir eğitim adil biraz gerektirir ve içine ajanlar direkt uygulanmasını içerir. 15,16 Çeşitli protokoller geliştirilmiştir solunum yolu güçlü bir yaralanmaya neden cerrahi müdahaleye gerek kalmadan trakea, 16,17,18,19,20 ancak bu yöntemler geniş bir alanını içermektedir kurtarma süreleri güçlü anestezik neden, pahalı ekipman kullanımı (yani., otoskop / laringoskop, ticari olarak mevcut prosedür panoları, fiber-optik doz ile ilgili olarak tel ve benzeri), ağız boşluğu içinde manipülasyon aşırı ve belirsizlik. Bu çalışma, bir araştırmacının hızla ucuz ve güvenilir bir şekilde çevre dokulara bakiye hasar sınırlı risk ile murin akciğer içine bir reaktif aşılamak sağlar entübasyon yoluyla uygulama nispeten kolay bir yöntem tarif etmektedir.

<table border="0" cellpadding="0" cellspacing="0" width="1528">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:481px;">Bleomycin For Injection, 30 units/vial</td>
			<td style="width:304px;">APP Pharmaceuticals, LLC</td>
			<td style="width:139px;">103720</td>
			<td style="width:604px;">For best results, BLM should be suspended in PBS, aliquoted, and stored as single use lyophilzed aliquots</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Blunt End Forceps</td>
			<td style="width:304px;">N/A</td>
			<td style="width:139px;">N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Tongue Depressor (i.e. bent Valleylab Blade Electrode, 2.4&#34;)&#160;</td>
			<td style="width:304px;">Covidien</td>
			<td>E1551G</td>
			<td>Before use, create a 45 degree bend 1.5 cm&#160; from the blade tip. A suitable depressor can also be created from any metal implement of similar dimensions.&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Exel Safelet Catheter 22G X 1&#34;</td>
			<td style="width:304px;">Exel International</td>
			<td>26746</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">1 mL Slip-tip Disposable Tuberculin Syringe (200/sp, 1600/ca)</td>
			<td style="width:304px;">BD</td>
			<td>309659</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">0.2ml Pipettor and Filter Tips</td>
			<td style="width:304px;">N/A</td>
			<td style="width:139px;">N/A</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:481px;">Fiber-Lite Illuminator. Model 181-1: Model 180 mated with Standard Dual Gooseneck illuminator</td>
			<td style="width:304px;">Dolan Jenner Industries, Inc.</td>
			<td style="width:139px;">181-1</td>
			<td>Lower output LED illuminators are not recommended as they fail to suficiently illuminate the trachea.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Intubation Board</td>
			<td style="width:304px;">N/A</td>
			<td style="width:139px;">N/A</td>
			<td>See Diagram 1.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Colored Label Tape: 0.5&#160;in. Wide</td>
			<td style="width:304px;">Fisherbrand</td>
			<td>15-901-15A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Oxygen&#160;</td>
			<td style="width:304px;">N/A</td>
			<td style="width:139px;">N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Phosphate-Buffered Saline, 1X</td>
			<td style="width:304px;">Corning</td>
			<td>21-040-CV</td>
			<td>Product should be sterile</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Non-Sterile Silk Black Braided Suture Spool, 91.4 m, Size 4-0</td>
			<td style="width:304px;">Harvard Apparatus</td>
			<td>517698</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Table Top Anesthesia Machine Isoflurane</td>
			<td style="width:304px;">Highland Medical Equipment</td>
			<td>N/A</td>
			<td>http://www.highlandmedical.net/</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Slide Top Induction Mouse Isoflurane Chamber</td>
			<td>MIP / Anesthesia Technologies</td>
			<td>AS-01-0530-SM</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">FORANE (isoflurane, USP) Liquid For Inhalation 100 mL&#160;</td>
			<td style="width:304px;">Baxter</td>
			<td>1001936040</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">&#160;Nanozoomer Digital Pathology system</td>
			<td style="width:304px;">Hamamatsu</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">IgM ELISA Quantification Kit&#160;</td>
			<td style="width:304px;">Bethyl Laboratories</td>
			<td style="width:139px;">E90-101</td>
			<td></td>
		</tr>
	</tbody>
</table>

an improved method for rapid intubation of the trachea in mice

Despite some anatomical and physiological differences, mouse models continue to be an essential tool for studying human lung disease. Bleomycin toxicity is a commonly used model to study both acute lung injury and fibrosis, and multiple methods have been developed for administering bleomycin (and other toxic agents) into the lungs. However, many of these approaches, such as transtracheal instillation, have inherent drawbacks, including the need for strong anesthetics and survival surgery. This paper reports a quick, reproducible method of intratracheal intubation that involves mild inhaled anesthesia, visualization of the trachea, and the use of a surrogate spirometer to confirm exposure. As a proof of concept, 8-12 week old C57BL/6 mice were administered either 2.0 U/kg of bleomycin or an equivalent volume of PBS, and both damage and fibrotic endpoints were measured post-exposure. This procedure allows researchers to treat a large cohort of mice in a relatively short period with little expense and minimal post-procedure care.

Entübe farenin kilo kaybı ve sıkıntı için günlük olarak izlenmiş ve% 2.5 2,2,2-tribromoetanol intraperitonal enjeksiyon yoluyla 4, 10 ya da 17 gün sonra feda edilmiştir. Bronkoalveoler lavaj (BAL) PBS üç yıkamadan içinde toplandı 21 başka bir yerde tarif edildiği gibi ve sağ akciğer,% 10 formalin sabit parafine gömülü ve Washington Histoloji ve Görüntüleme Çekirdek 22 Üniversitesi tarafından Masson trikrom ile boyandı....

Watch this Scientific Journal Video about An Improved Method for Rapid Intubation of the Trachea in Mice at JoVE.com

An Improved Method for Rapid Intubation of the Trachea in Mice

Farelerde Trakea hızlı intübasyon için geliştirilmiş bir yöntem

Bu makalede, entübasyon yoluyla fare trakea içine doğrudan bleomisin yönetmek için hızlı ve basit bir yöntem sunar. Bu yöntemin en önemli avantajları usta son derece tekrarlanabilir kolay ve özel ekipman ya da uzun iyileşme süreleri gerektirmez vardır.

Despite some anatomical and physiological differences, mouse models continue to be an essential tool for studying human lung disease. Bleomycin toxicity ...

an-improved-method-for-rapid-intubation-of-the-trachea-in-mice

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

25.0K Görüntüleme.  University of Washington.  Bu makalede, entübasyon yoluyla fare trakea içine doğrudan bleomisin yönetmek için hızlı ve basit bir yöntem sunar. Bu yöntemin en önemli avantajları usta son derece tekrarlanabilir kolay ve özel ekipman ya da uzun iyileşme süreleri gerektirmez vardır. 

Video: An Improved Method for Rapid Intubation of the Trachea in Mice

Guidelines for Elective Pediatric Fiberoptic Intubation

Fiberoptic intubation in pediatric patients is often required especially in difficult airways of syndromic patients i.e. Pierre Robin Syndrome. Small babies will desaturate very quickly if ventilation is interrupted mainly to high metabolic rate. We describe guidelines to perform a safe fiberoptic intubation while maintaining spontaneous breathing throughout the procedure. Steps requiring the use of propofol pump, fentanyl, glycopyrrolate, red rubber catheter, metal insuflation hook, afrin, lubricant and lidocaine spray are shown.

We describe guidelines to perform a safe and efficient elective fiberoptic intubation in pediatric patients while maintaining spontaneous ventilation.

Seçmeli Pediatrik Fiberoptik Entübasyon için rehber

Fiberoptic intubation in pediatric patients is often required especially in difficult airways of syndromic patients i.e. Pierre Robin Syndrome. Small ...

Tıp

Orthotopic Aortic Transplantation in Mice for the Study of Vascular Disease

Vascular procedures involving anastomoses in the mouse are generally thought to be difficult and highly dependent on the skill of the individual surgeon. This is largely true, but there are a number of important principles that can reduce the difficulty of these procedures and enhance reproducibility. Orthotopic aortic transplantation is an excellent procedure in which to learn these principles because it involves only two end-to-end anastomoses, but requires good suturing technique and handling of the vessels for consistent success. This procedure begins with the procurement of a length of abdominal aorta from a donor animal, followed by division of the native aorta in the recipient. The procured aorta is then placed between the divided ends of the recipient aorta and sutured into place using end-to-end anastomoses. To accomplish this objective successfully requires a high degree of concentration, good tools, a steady hand, and an appreciation of how easily the vasculature of a mouse can be damaged, resulting in thrombosis. Learning these important principles is what occupies most of the beginner's time when learning microsurgery in small rodents. Throughout this protocol, we refer to these important points. This model can be used to study vascular disease in a variety of different experimental systems1-8. In the context shown here, it is most often used for the study of post-transplant vascular disease, a common long-term complication of solid organ transplantation in which intimal hyperplasia occurs within the allograft. The primary advantage of the model is that it facilitates quantitative morphometric analyses and the transplanted vessel lies contiguous to the endogenous vessel, which can serve as an additional control9. The technique shown here is most often used for mice weighing 18-25 grams. We have accumulated most of our experience using the C57BL/6J, BALB/cJ, and C3H/HeJ strains.

We describe a technique in which a section of the abdominal aorta from a mouse is transplanted orthotopically to just below the renal arteries in an allogeneic or syngeneic recipient. This technique can be useful in studies in which transplantation of large arteries of uniform size is deemed advantageous.

Damar Hastalığı Çalışmaları Farelerde Ortotopik Aort Nakli

Vascular procedures involving anastomoses in the mouse are generally thought to be difficult and highly dependent on the skill of the individual surgeon. ...

Endotracheal Intubation in Mice via Direct Laryngoscopy Using an Otoscope

Mice, both wildtype and transgenic, are the principal mammalian model in biomedical research currently. Intubation and mechanical ventilation are necessary for whole animal experiments that require surgery under deep anesthesia or measurements of lung function. Tracheostomy has been the standard for intubating the airway in these mice to allow mechanical ventilation. Orotracheal intubation has been reported but has not been successfully used in many studies because of the substantial technical difficulty or a requirement for highly specialized and expensive equipment. Here we report a technique of direct laryngoscopy using an otoscope fitted with a 2.0 mm speculum and using a 20 G&#160;intravenous catheter as an endotracheal tube. We have used this technique extensively and reliably to intubate and conduct accurate assessments of lung function in mice. This technique has proven safe, with essentially no animal loss in experienced hands. Moreover, this technique can be used for repeated studies of mice in chronic models.

Endotracheal Intubation in Mice via Direct Laryngoscopy Using an Otoscope

We have developed a simple, reliable, and relatively inexpensive method for endotracheal intubation in mice via direct laryngoscopy using an otoscope with a 2.0 mm speculum. This technique is atraumatic and can be used for repeated measurements in chronic experiments. We find it superior to tracheostomy or previously reported nonsurgical techniques.

Farelerde Endotrakeal Entübasyon<em&gt; Via</emBir Otoskop Kullanma&gt; Doğrudan Larengoskopi

Mice, both wildtype and transgenic, are the principal mammalian model in biomedical research currently. Intubation and mechanical ventilation are ...

A Simple Method of Mouse Lung Intubation

A simple procedure to intubate mice for pulmonary function measurements would have several advantages in longitudinal studies with limited numbers or expensive animal. One of the reasons that this is not done more routinely is that it is relatively difficult, despite there being several published studies that describe ways to achieve it. In this paper we demonstrate a procedure that eliminates one of the major hurdles associated with this intubation, that of visualizing the trachea during the entire time of intubation. The approach uses a 0.5 mm fiberoptic light source that serves as an introducer to direct the intubation cannula into the mouse trachea. We show that it is possible to use this procedure to measure lung mechanics in individual mice over a time course of at least several weeks. The technique can be set up with relatively little expense and expertise, and it can be routinely accomplished with relatively little training. This should make it possible for any laboratory to routinely carry out this intubation, thereby allowing longitudinal studies in individual mice, thereby minimizing the number of mice needed and increasing the statistical power by using each mouse as its own control.

This paper describes a striaghforward and efficient method of intubating mice for pulmonary function measurements or pulmonary instillation, that allows the mice to recover and be studied at later times. The procedure involves an inexpensive fiberoptic light source that directly illuminates the trachea.

Fare Akciğer Entübasyonun Basit Bir Yöntem

A  simple procedure to intubate mice for pulmonary function measurements would  have several advantages in longitudinal studies with limited numbers or  ...

Improved Method for the Establishment of an In Vitro Blood-Brain Barrier Model Based on Porcine Brain Endothelial Cells

The aim of this protocol presents an optimized procedure for the purification and cultivation of pBECs and to establish in vitro blood-brain barrier (BBB) models based on pBECs in mono-culture (MC), MC with astrocyte-conditioned medium (ACM), and non-contact co-culture (NCC) with astrocytes of porcine or rat origin. pBECs were isolated and cultured from fragments of capillaries from the brain cortices of domestic pigs 5-6 months old. These fragments were purified by careful removal of meninges, isolation and homogenization of grey matter, filtration, enzymatic digestion, and centrifugation. To further eliminate contaminating cells, the capillary fragments were cultured with puromycin-containing medium. When 60-95% confluent, pBECs growing from the capillary fragments were passaged to permeable membrane filter inserts and established in the models. To increase barrier tightness and BBB characteristic phenotype of pBECs, the cells were treated with the following differentiation factors: membrane permeant 8-CPT-cAMP (here abbreviated cAMP), hydrocortisone, and a phosphodiesterase inhibitor, RO-20-1724 (RO). The procedure was carried out over a period of 9-11 days, and when establishing the NCC model, the astrocytes were cultured 2-8 weeks in advance. Adherence to the described procedures in the protocol has allowed the establishment of endothelial layers with highly restricted paracellular permeability, with the NCC model showing an average transendothelial electrical resistance (TEER) of 1249 &#177; 80 &#937; cm2, and paracellular permeability (Papp) for Lucifer Yellow of 0.90 10-6 &#177; 0.13 10-6 cm sec-1 (mean &#177; SEM, n=55). Further evaluation of this pBEC phenotype showed good expression of the tight junctional proteins claudin 5, ZO-1, occludin and adherens junction protein p120 catenin. The model presented can be used for a range of studies of the BBB in health and disease and, with the highly restrictive paracellular permeability, this model is suitable for studies of transport and intracellular trafficking.

Improved Method for the Establishment of an In Vitro Blood-Brain Barrier Model Based on Porcine Brain Endothelial Cells

The aim of the protocol is to present an optimized procedure for the establishment of an in vitro blood-brain barrier (BBB) model based on primary porcine brain endothelial cells (pBECs). The model shows high reproducibility, high tightness, and is suitable for studies of transport and intracellular trafficking in drug discovery.

Yöntem bir Vitro kan - beyin bariyerini modeli domuz beyin endotel hücreleri temel alan kurulması için geliştirilmiş

The aim of this protocol presents an optimized procedure for the purification and cultivation of pBECs and to establish in vitro blood-brain barrier (BBB) ...

Assessment and Characterization of Hyaloid Vessels in Mice

In the eye, the embryonic hyaloid vessels nourish the developing lens and retina and regress when the retinal vessels develop. Persistent or failed regression of hyaloid vessels can be seen in diseases such as persistent hyperplastic primary vitreous (PHPV), leading to an obstructed light path and impaired visual function. Understanding the mechanisms underlying the hyaloid vessel regression may lead to new molecular insights into the vascular regression process and potential new ways to manage diseases with persistent hyaloid vessels. Here we describe the procedures for imaging hyaloid in live mice with optical coherence tomography (OCT) and fundus fluorescein angiography (FFA) and a detailed technical protocol of isolating and flat-mounting hyaloid ex vivo for quantitative analysis. Low-density lipoprotein receptor-related protein 5 (LRP5) knockout mice were used as an experimental model of persistent hyaloid vessels, to illustrate the techniques. Together, these techniques may facilitate a thorough assessment of hyaloid vessels as an experimental model of vascular regression and studies on the mechanism of persistent hyaloid vessels.

This protocol describes both in vivo and ex vivo methods to fully visualize and characterize hyaloid vessels, a model of vascular regression in mouse eyes, using optical coherence tomography and fundus fluorescein angiography for the live imaging and ex vivo isolation and subsequent flat mount of hyaloid for quantitative analysis.

Farelerde hyaloid damarlarının değerlendirilmesi ve karakterize edilmesi

In the eye, the embryonic hyaloid vessels nourish the developing lens and retina and regress when the retinal vessels develop. Persistent or failed ...

A Revised Method for Inducing Secondary Lymphedema in the Hindlimb of Mice

Animal models are of paramount importance in the research of lymphedema in order to understand the pathophysiology of the disease but also to explore potential treatment options. This mouse model allows researchers to induce significant lymphedema lasting at least 8 weeks. Lymphedema is induced using a combination of fractioned radiotherapy and surgical ablation of lymphatics. This model requires that the mice get a dose of 10 Gray (Gy) radiation before and after surgery. The surgical part of the model involves ligation of three lymph vessels and extraction of two lymph nodes from the mouse hindlimb. Having access to microsurgical tools and a microscope is essential, due to the small anatomical structures of mice. The advantage of this model is that it results in statistically significant lymphedema, which provides a good basis for evaluating different treatment options. It is also a great and easily available option for microsurgical training. The limitation of this model is that the procedure can be time consuming, especially if not practiced in advance. The model results in objectively quantifiable lymphedema in mice, without causing severe morbidity and has been tested in three separate projects.

This animal model enables researchers to induce statistically significant secondary lymphedema in the hindlimb of mice, lasting at least 8 weeks. The model can be used to study the pathophysiology of lymphedema and to investigate novel treatment options.

Farelerin Hindlimb Sekonder Lenfödem Indükleme için Gözden Geçirilmiş Bir Yöntem

Animal models are of paramount importance in the research of lymphedema in order to understand the pathophysiology of the disease but also to explore ...

Repeated Orotracheal Intubation in Mice

The literature describes several methods for mouse intubation that either require visualization of the glottis through the oral cavity or incision in the ventral neck for direct confirmation of cannula placement in the trachea. The relative difficulty or the tissue trauma induced to the subject by such procedures can be an impediment to an investigator&#8217;s ability to perform longitudinal studies. This article illustrates a technique in which physical manipulation of the mouse following the use of a depilatory to remove hair from the ventral neck permits transcutaneous visualization of the trachea for orotracheal intubation regardless of degree of skin pigmentation. This method is innocuous to the subject and easily achieved with a limited understanding of murine anatomy. This refined approach facilitates repeated intubation, which may be necessary for monitoring progression of disease or instillation of treatments. Using this method may result in a reduction of the number of animals and technical skill required to measure lung function in mouse models of respiratory disease.

The goal of this article is to describe a refined method of intubation of the laboratory mouse. The method is noninvasive and, therefore, ideal for studies that require serial monitoring of respiratory function and/or instillation of treatments into the lung.

Farelerde Tekrarlanan Orotracheal Entübasyon

The literature describes several methods for mouse intubation that either require visualization of the glottis through the oral cavity or incision in the ...

Semi-Minimal Invasive Method to Induce Myocardial Infarction in Rats and the Assessment of Cardiac Function by an Isolated Working Heart System

Myocardial infarction (MI) remains the main contributor to morbidity and mortality worldwide. Therefore, research on this topic is mandatory. An easily and highly reproducible MI induction procedure is required to obtain further insight and better understanding of the underlying pathological changes. This procedure can also be used to evaluate the effects or potency of new and promising treatments (as drugs or interventions) in acute MI, subsequent remodeling and heart failure (HF). After intubation and pre-operative preparation of the animal, an anesthetic protocol with isoflurane was performed, and the surgical procedure was conducted&#160;quickly. Using a minimally invasive approach, the left anterior descending artery (LAD) was located and occluded by a ligature. The occlusion can be performed acutely for subsequent reperfusion (ischemia/reperfusion injury). Alternatively, the vessel can be ligated permanently to investigate the development of chronic MI, remodeling or HF. Despite common pitfalls, the drop-out rates are minimal. Various treatments such as remote ischemic conditioning can be examined for their cardioprotective potential pre-, peri- and post-operatively. The post-operative recovery was quick as the anesthesia was precisely controlled and the duration of the operation was short. Post-operative analgesia was administered for three days. The minimally invasive procedure reduces the risk of infection and inflammation. Furthermore, it facilitates rapid recovery. The &#8220;working heart&#8221; measurements were performed ex vivo and enabled precise control of preload, afterload and flow. This procedure requires specific equipment and training for adequate performance. This manuscript provides a detailed step-by-step introduction for conducting these measurements.

This article presents an efficient method to perform myocardial ischemia and subsequent chronic reperfusion in rats using a minimally invasive approach. In addition, left ventricular hemodynamic function of rats is assessed by echocardiography and isolated working heart methods.&#160;

Sıçanlarda Miyokard İnfarktüsü İndükleyen Yarı-Minimal İnvaziv Yöntem ve İzole Çalışan Kalp Sistemi Ile Kardiyak Fonksiyonun Değerlendirilmesi

Myocardial infarction (MI) remains the main contributor to morbidity and mortality worldwide. Therefore, research on this topic is mandatory. An easily ...

A Modified Simple Method for Induction of Myocardial Infarction in Mice

Myocardial infarction (MI) represents one of the leading causes of death. MI models are widely used for investigating the pathomechanisms of post-MI remodeling and evaluation of novel therapeutics. Different methods (e.g., isoproterenol treatment, cryoinjury, coronary artery ligation, etc.) have been used to induce MI. Compared with isoproterenol treatment and cryoinjury, coronary artery ligation may better reflect the ischemic response and chronic remodeling after MI. However, traditional methods for coronary ligation in mice are technically challenging. The current study describes a simple and efficient process for induction of MI in mice with readily available materials. The mouse chest skin was cut open under stable anesthesia. The heart was immediately externalized through the intercostal space after blunt separation of the pectoralis major and pectoralis minor. The left anterior descending branch (LAD) was ligated with a 6-0 suture 3 mm from its origin. Following LAD ligation, staining with 2,3,5-Triphenyltetrazolium chloride (TTC) indicated successful induction of MI and temporal changes of post-MI scar size. Meanwhile, survival analysis results showed overt mortality within 7 days after MI, mainly due to cardiac rupture. Moreover, post-MI echocardiographic assessment demonstrated successful induction of contractile dysfunction and ventricular remodeling. Once mastered, an MI model can be established in mice within 2-3 min with readily available materials.

Under adequate anesthesia, the mouse heart was externalized through the intercostal space, and myocardial infarction was successfully induced by ligating the left anterior descending artery (LAD) using materials readily available in most laboratories.

Farelerde Miyokard Enfarktüsünün İndüksiyonu için Değiştirilmiş Basit Bir Yöntem

Myocardial infarction (MI) represents one of the leading causes of death. MI models are widely used for investigating the pathomechanisms of post-MI ...