Soutenu par le NIH HL-10342.

1. Préparation de la procédure Il faut d&#39;abord obtenir et préparer les éléments suivants: <ol><li> La canule. Pour l&#39;intubation des souris g 20-35, nous utilisons un pouce 1 ou 1,5 de long, calibre 20 IV cathéter (BD Insylte, Sparks, MD ou Jelco Optiva, Carlsbad, CA). Un nouveau cathéter stérile peut être utilisé pour chaque souris, mais cathéters peut également être réutilisé après stérilisation par immersion dans de l&#39;éthanol 70% pendant une nuit. Bien que ni le pharynx, ni trachée de la souris est stériles procédures propreté appropriées, y compris l&#39;utilisation de gants stériles et des instruments, doivent être suivies. </li><li> Le câble à fibres optiques. Nous utilisons ≈ 70 cm de câble de 0,5 mm optique d&#39;Edmund Optics, mais la longueur n&#39;est pas critique. Il est important de s&#39;assurer que la fibre a son bord lissé, car après avoir coupé le câble à la longueur avec une lame de rasoir, le bord gauche est relativement forte, et il ne prend pas beaucoup d&#39;effort pour percer la paroi de la trachée. Cependant, il iest très facile à lisser ce bord en maintenant la fibre d&#39;environ 2 cm de l&#39;extrémité, puis faisant des petits cercles pendant quelques secondes avec les bords de la pointe de toucher un morceau de papier abrasif 1000 (voir la démonstration dans la vidéo et la figure 1 de Macdonald et al. 10). L&#39;autre extrémité est insérée à travers un bouchon de caoutchouc. Le plus simple est d&#39;abord faire pousser une aiguille de calibre 18 à travers le bouchon, insérer la fibre optique à travers le trou d&#39;aiguille, puis retirer l&#39;aiguille. Le bouchon en caoutchouc est relié à une source de lumière halogène de 150 watts (par exemple NCL-150, Volpi USA, ou toute autre source de lumière ou même moins de 150 watts). Il est important de s&#39;assurer d&#39;utiliser un bouchon en caoutchouc de silicone (ou autre matériau résistant à la chaleur d&#39;autres), depuis caoutchouc ordinaire ou en liège peut brûler lorsqu&#39;il est situé si près de la source chaude lumière. </li></ol> 2. Exécution de l&#39;intubation <ol><li> Voir les figures 1 et 2. Insérez câble à fibre optique trâce à un court morceau de tuyau en caoutchouc de silicone (≈ 0,8 mm ID x 4 mm de diamètre, Cole-Palmer, EW-96410-13). Attachez ce tube en caoutchouc assez serré, tout en permettant au câble à fibre optique à régler. Insérer le tube de silicone sans jeu dans l&#39;extrémité de la canule Luer fixe la position du câble à fibres optiques à l&#39;intérieur de la canule. Ajuster la position du câble à fibres optiques de la sorte qu&#39;il s&#39;étend à travers la canule ≈ 4 mm en avant de la pointe de la canule. </li><li> Placez la souris anesthésiée sur un support vertical, suspendu par ses incisives supérieures (figure 3). La plupart des chercheurs trouver la meilleure visualisation avec la face ventrale de la souris se faisant face. Très tirez doucement sur la langue et tenir avec le pouce et l&#39;index. Le doigt du milieu est placé entre le col et le support en plastique. Traction sur la languette avec le pouce et l&#39;index est utilisé pour ouvrir la bouche, et de redresser le chemin intubation, l&#39;angle de la tête est réglée avec le doigt du milieu derrière le coureprésenté sur la figure 3. </li><li> En utilisant le câble de fibre optique en tant que source de lumière et d&#39;introduction, le pousser à travers les cordes vocales visualisées. Si les cordons ne sont pas visibles, tirez doucement sur la langue plus difficile avec le doigt du milieu comme support. Lorsqu&#39;elle est insérée, faire avancer la canule ≈ 5 mm de plus. Puis, en faisant très attention de ne pas déplacer la canule, retirer le câble de fibre optique. Allongez la souris vers le bas et fixer la canule avec un morceau de ruban adhésif et de soutenir le moyeu de canule sur un morceau de pâte à modeler (pâte à modeler), comme le montre la figure 4. </li><li> La procédure de l&#39;étape 3 ne peut pas être facilement enseigné ou même montré, car il s&#39;agit d&#39;une opération en solo. Cependant, par des ajustements subtils de la traction sur la languette et le soutien derrière la tête presque tous ceux qui tentent ce bientôt trouver la bonne façon de positionner la souris pour visualiser les cordes vocales. </li></ol>

<ol>
	<li>Brown, R. H., Walters, D. M., Greenberg, R. S., 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=A+method+of+endotracheal+intubation+and+pulmonary+functional+assessment+for+repeated+studies+in+mice.">A method of endotracheal intubation and pulmonary functional assessment for repeated studies in mice.</a> J. Appl. Physiol. 87, 2362-2365 (1999).</li><li>Walters, D. M., Wills-Karp, M., 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=Assessment+of+cellular+profile+and+lung+function+with+repeated+bronchoalveolar+lavage+in+individual+mice.">Assessment of cellular profile and lung function with repeated bronchoalveolar lavage in individual mice.</a> Physiol. Genomics. 2, 29-36 (2000).</li><li>Rivera, B., Miller, S., Brown, E., Price, R. <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+method+for+endotracheal+intubation+of+mice+and+rats+used+in+imaging+studies.">A novel method for endotracheal intubation of mice and rats used in imaging studies.</a> Contemporary Topics in Laboratory Animal Science / American Association for Laboratory Animal Science. 44, 52-55 (2005).</li><li>Hamacher, 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=Microscopic+wire+guide-based+orotracheal+mouse+intubation:+description,+evaluation+and+comparison+with+transillumination.">Microscopic wire guide-based orotracheal mouse intubation: description, evaluation and comparison with transillumination.</a> Laboratory Animals. 42, 222-230 (2008).</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> Laboratory Animals. 41, 128-135 (2007).</li><li>Zhao, 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=A+technique+for+retrograde+intubation+in+mice.">A technique for retrograde intubation in mice.</a> Lab Animal. 35, 39-42 (2006).</li><li>Vergari, 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=A+new+method+of+orotracheal+intubation+in+mice.">A new method of orotracheal intubation in mice.</a> European Review for Medical and Pharmacological Sciences. 8, 103-106 (2004).</li><li>Vergari, A., Polito, A., Musumeci, M., Palazzesi, S., Marano, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Video-assisted+orotracheal+intubation+in+mice.">Video-assisted orotracheal intubation in mice.</a> Laboratory Animals. 37, 204-206 (2003).</li><li>Hastings, R. H., Summers-Torres, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+Laryngoscopy+in+Mice.">Direct Laryngoscopy in Mice.</a> Contemporary Topics in Laboratory Animal Science / American Association for Laboratory Animal Science. 38, 33-35 (1999).</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. 106, 984-987 (2009).</li><li>Ewart, S. L., Levitt, R. C., 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=Respiratory+system+mechanics+in+mice+measured+by+end-inflation+occlusion.">Respiratory system mechanics in mice measured by end-inflation occlusion.</a> Journal of Applied Physiology. 79, 560-566 (1995).</li><li>MacDonald, K. D., McKenzie, K. R., Mitzner, W., Zeitlin, P. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lung+Mechanics+in+Heterozygous+CF+Mice+after+Repeated+LPS+Dosing.">Lung Mechanics in Heterozygous CF Mice after Repeated LPS Dosing.</a> Am. J. Respir. Crit. Care Med. 175 (4), A930 (2007).</li></ol>

Aucun des auteurs n&#39;a de conflit d&#39;intérêt à déclarer.

La procédure décrite ici présente plusieurs avantages. D&#39;abord l&#39;appareil est simple et relativement peu coûteux .. La fabrication de l&#39;appareil ne nécessite pas d&#39;outils spéciaux ou de l&#39;équipement coûteux. L&#39;utilisation d&#39;un cathéter introduit qui est également la source de lumière signifie que l&#39;on ne perd jamais de vue l&#39;ouverture de la trachée comme l&#39;introducteur s&#39;approche de l&#39;ouverture trachéale. L&#39;utilisation d&#39;un dispositif d&#39;introductio...

En 1999, Brown et al. Publié un article décrivant une méthode pour l&#39;intubation du poumon 1 souris. Une telle technique a une utilité considérable en faisant fonction de répétition ou lavage broncho-alvéolaire pulmonaire chez la souris individuelles dans les études longitudinales 2. Depuis que le papier d&#39;origine, il ya eu plusieurs autres documents qui ont décrit les différentes approches à 3-9 intubation souris. Bien que toutes ces méthodes peuvent être utilisées avec succès, ils exigent habituellement une formation considérable ou de coût. Un des principaux problèmes avec intubation par exemple, c&#39;est que l&#39;approche de la canule d&#39;intubation se rapproche de la trachée attente d&#39;insertion, la canule elle-même bloque la lumière et donc la visualisation de l&#39;endroit où il doit aller. Ainsi, l&#39;insertion devient aveugle au moment le plus critique. Dans cet article, nous montrons comment simple et peu coûteuse d&#39;éliminer ce problème de visualisation, assurant ainsi une intubation réussie avec relativement peu de formation ouexpérience.

<table border="1"><tbody><tr><td> Matériel </td><td> Entreprise </td><td> Catalogue </td><td> Commenter </td></tr><tr><td> Canule d&#39;intubation </td><td> BD Insylte, Sparks, MD ou Jelco Optiva, Carlsbad, CA </td><td></td><td> 1-po de longueur, de calibre 20 par voie intraveineuse (IV) du cathéter </td></tr><tr><td> Câble à fibres optiques </td><td> Edmund Optics, Barrington, NJ </td><td> # NT02-542 </td><td> Environ 2 pi longueur de 0,5 mm de fibre optique (fibre plastique communication grade). Le bord de l&#39;extrémité de la fibre qui est inséré dans la trachée doit être légèrement arrondi en maintenant la fibre ≈ 2 cm de l&#39;extrémité, puis faisant de petits cercles tout en faisant glisser la pointe pendant quelques secondes sur 1000 emory papier abrasif. </td></tr><tr><td> Source de lumière </td><td> Volpi </td><td> NCL-150 </td><td> Bien que nous utilisions une source de 150 W lampe halogène, une source de lumière équivalente, même avec une puissance beaucoup plus faiblepeuvent être utilisées. </td></tr><tr><td> Bouchon en caoutchouc de silicone </td><td></td><td></td><td> Un bouchon de caoutchouc de silicone n ° 1 correspond à la source de lumière utilisée dans cette démonstration. Différentes sources de lumière peuvent exiger une taille différente. </td></tr><tr><td> Petit tube en caoutchouc silicone </td><td> Cole-Palmer </td><td> EW-96410-13 </td><td> Un morceau ≈ 1,5 cm de tube en caoutchouc de silicone (0,8 diamètre intérieur de 4 mm de diamètre extérieur) </td></tr><tr><td> Montant de support incliné </td><td></td><td></td><td> La nôtre est constucted en plexiglas, mais toute matière à laquelle un fil ou câble peut être fixé à tenir la souris à un angle presque vertical peut être utilisé. </td></tr></tbody></table>

a simple method of mouse lung intubation

Une procédure simple pour intuber souris pour les mesures de la fonction pulmonaire aurait plusieurs avantages dans les études longitudinales avec un nombre limité d&#39;animaux ou coûteux. Une des raisons pour lesquelles cela n&#39;est pas fait de façon plus routinière, c&#39;est qu&#39;il est relativement difficile, malgré l&#39;existence de plusieurs études publiées qui décrivent les moyens pour y parvenir. Dans cet article, nous démontrons une procédure qui élimine l&#39;un des obstacles majeurs associés à ce intubation, qui permet de visualiser la trachée pendant toute la durée de l&#39;intubation. La méthode utilise une source de lumière à fibre optique 0,5 mm qui sert d&#39;introduction pour diriger la canule d&#39;intubation dans la trachée de souris. Nous montrons qu&#39;il est possible d&#39;utiliser cette procédure pour mesurer la mécanique du poumon chez les souris individuelles sur un parcours de temps d&#39;au moins plusieurs semaines. La technique peut être mis en place avec relativement peu de frais et d&#39;expertise, et il peut être systématiquement réalisé avec relativement peu de formation. Cela devrait make il possible pour tout laboratoire réalise ce intubation, ce qui permet des études longitudinales chez des souris individuelles, ce qui réduit le nombre de souris nécessaires et en augmentant la puissance statistique en utilisant chaque souris comme son propre témoin.

Comme une évaluation de la méthode, nous avons utilisé quatre mâles âgés de 20 semaines BALB / c avec un poids moyen (± ET) de 27,7 ± 0,40 g. Ils ont été étudiés sur cinq semaines consécutives, où la résistance pulmonaire a été mesurée en utilisant un système tel que décrit précédemment 11. Chaque souris a été anesthésiée avec de la kétamine (100 mg / g de poids) et de xylazine (15 mg / g de poids) dans une solution saline par injection IP. Ils ont ensuite été intubé comme décri...

Watch this Scientific Journal Video about A Simple Method of Mouse Lung Intubation at JoVE.com

A Simple Method of Mouse Lung Intubation

Le présent document décrit une méthode striaghforward et efficace de l&#39;intubation des souris pour les mesures de la fonction pulmonaire ou l&#39;instillation pulmonaire, qui permet aux souris de récupérer et être étudié à temps postérieurs. La procédure implique une source de lumière à fibre optique peu coûteux qui éclaire directement la trachée.

Une méthode simple d&#39;intubation poumon de souris

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

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Medicine

50.5K vues.  Johns Hopkins Bloomberg School of Public Health. Le présent document décrit une méthode striaghforward et efficace de l'intubation des souris pour les mesures de la fonction pulmonaire ou l'instillation pulmonaire, qui permet aux souris de récupérer et être étudié à temps postérieurs. La procédure implique une source de lumière à fibre optique peu coûteux qui éclaire directement la trachée.

Vidéo: A Simple Method of Mouse Lung Intubation

A Model of Disturbed Flow-Induced Atherosclerosis in Mouse Carotid Artery by Partial Ligation and a Simple Method of RNA Isolation from Carotid Endothelium

Despite the well-known close association, direct evidence linking disturbed flow to atherogenesis has been lacking. We have recently used a modified version of carotid partial ligation methods [1,2] to show that it acutely induces low and oscillatory flow conditions, two key characteristics of disturbed flow, in the mouse common carotid artery. Using this model, we have provided direct evidence that disturbed flow indeed leads to rapid and robust atherosclerosis development in Apolipoprotein E knockout mouse [3]. We also developed a method of endothelial RNA preparation with high purity from the mouse carotid intima [3]. Using this mouse model and method, we found that partial ligation causes endothelial dysfunction in a week, followed by robust and rapid atheroma formation in two weeks in a hyperlipidemic mouse model along with features of complex lesion formation such as intraplaque neovascularization by four weeks. This rapid in vivo model and the endothelial RNA preparation method could be used to determine molecular mechanisms underlying flow-dependent regulation of vascular biology and diseases. Also, it could be used to test various therapeutic interventions targeting endothelial dysfunction and atherosclerosis in considerably reduced study duration.

This describes a partial carotid ligation surgery, which causes disturbed flow conditions and subsequent atherosclerosis development (in two weeks) with intraplaque neo-vascularization (in four weeks) in the mouse common carotid artery. We also describe a novel method of RNA isolation from the carotid intima, providing high purity endothelial RNA.

Un modèle de Disturbed flux athérosclérose induite chez la souris artère carotide par ligature partielle et une méthode simple d&#39;isolement d&#39;ARN de l&#39;endothélium carotidienne

Despite the well-known close association, direct evidence linking disturbed flow to atherogenesis has been lacking. We have recently used a modified ...

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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.

Lignes directrices pour l&#39;intubation fibroscopique pédiatrique élective

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

A Simple Guide Screw Method for Intracranial Xenograft Studies in Mice

The grafting of human tumor cells into the brain of immunosuppressed mice is an established method for the study of brain cancers including glioblastoma (glioma) and medulloblastoma. The widely used stereotactic approach only allows for the injection of a single animal at a time, is labor intensive and requires highly specialized equipment. The guide screw method, initially developed by Lal et al.,1 was developed to eliminate cumbersome stereotactic procedures. We now describe a modified guide screw approach that is rapid and exceptionally safe; both of which are critical ethical considerations. Notably, our procedure now incorporates an infusion pump that allows up to 10 animals to be simultaneously injected with tumor cells.
To demonstrate the utility of this procedure, we established human U87MG glioma cells as intracranial xenografts in mice, which were then treated with AMG102; a fully human antibody directed to HGF/scatter factor currently undergoing clinical evaluation2-5. Systemic injection of AMG102 significantly prolonged the survival of all mice with intracranial U87MG xenografts and resulted in a number of complete cures. 
This study demonstrates that the guide screw method is an inexpensive, highly reproducible approach for establishing intracranial xenografts. Furthermore, it provides a relevant physiological model for validating novel therapeutic strategies for the treatment of brain cancers.

In order to evaluate novel therapeutic paradigms for the treatment of glioma, physiological relevant models are essential. We utilize an implantable guide screw procedure for establishment of intracranial xenograft models that is more rapid and safer than stereotactic approaches.

Une méthode simple guide de vis pour intracrânienne études de xénogreffe chez la souris

The grafting of human tumor cells into the brain of immunosuppressed mice is an established method for the study of brain cancers including glioblastoma ...

Noninvasive Intratracheal Intubation to Study the Pathology and Physiology of Mouse Lung

The use of a model that mimics the condition of lung diseases in humans is critical for studying the pathophysiology and/or etiology of a particular disease and for developing therapeutic intervention. With the increasing availability of knockout and transgenic derivatives, together with a vast amount of genetic information, mice provide one of the best models to study the molecular mechanisms underlying the pathology and physiology of lung diseases. Inhalation, intranasal instillation, intratracheal instillation, and intratracheal intubation are the most widely used techniques by a number of investigators to administer materials of interest to mouse lungs. There are pros and cons for each technique depending on the goals of a study. Here a noninvasive intratracheal intubation method that can directly deliver exogenous materials to mouse lungs is presented. This technique was applied to administer bleomycin to mouse lungs as a model to study pulmonary fibrosis.

The use of a model that mimics the condition of lung diseases in humans is critical for studying the pathophysiology and/or etiology of a particular disease and for developing therapeutic intervention. Here a noninvasive intratracheal intubation method that can directly deliver exogenous materials to mouse lungs is presented.

Non invasive Intratrachéal intubation pour étudier la pathologie et la physiologie des souris poumon

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Implantation of Fibrin Gel on Mouse Lung to Study Lung-specific Angiogenesis

Recent significant advances in stem cell research and bioengineering techniques have made great progress in utilizing biomaterials to regenerate and repair damage in simple tissues in the orthopedic and periodontal fields. However, attempts to regenerate the structures and functions of more complex three-dimensional (3D) organs such as lungs have not been very successful because the biological processes of organ regeneration have not been well explored. It is becoming clear that angiogenesis, the formation of new blood vessels, plays key roles in organ regeneration. Newly formed vasculatures not only deliver oxygen, nutrients and various cell components that are required for organ regeneration but also provide instructive signals to the regenerating local tissues. Therefore, to successfully regenerate lungs in an adult, it is necessary to recapitulate the lung-specific microenvironments in which angiogenesis drives regeneration of local lung tissues. Although conventional in vivo angiogenesis assays, such as subcutaneous implantation of extracellular matrix (ECM)-rich hydrogels (e.g., fibrin or collagen gels or Matrigel - ECM protein mixture secreted by Engelbreth-Holm-Swarm mouse sarcoma cells), are extensively utilized to explore the general mechanisms of angiogenesis, lung-specific angiogenesis has not been well characterized because methods for orthotopic implantation of biomaterials in the lung have not been well established. The goal of this protocol is to introduce a unique method to implant fibrin gel on the lung surface of living adult mouse, allowing for the successful recapitulation of host lung-derived angiogenesis inside the gel. This approach enables researchers to explore the mechanisms by which the lung-specific microenvironment controls angiogenesis and alveolar regeneration in both normal and pathological conditions. Since implanted biomaterials release and supply physical and chemical signals to adjacent lung tissues, implantation of these biomaterials on diseased lung can potentially normalize the adjacent diseased tissues, enabling researchers to develop new therapeutic approaches for various types of lung diseases.

Recapitulation of the organ-specific microenvironment, which stimulates local angiogenesis, is indispensable for successful regeneration of damaged tissues. This report demonstrates a novel method to implant fibrin gels on the lung surface of living mouse in order to explore how the lung-specific microenvironment modulates angiogenesis and alveolar regeneration in adult mouse.

Implantation de fibrine Gel sur la souris Lung pour étudier l&#39;angiogenèse pulmonaire spécifique

Recent significant advances in stem cell research and bioengineering techniques have made great progress in utilizing biomaterials to regenerate and ...

Intubation-mediated Intratracheal (IMIT) Instillation: A Noninvasive, Lung-specific Delivery System

Respiratory disease studies typically involve the use of murine models as surrogate systems. However, there are significant physiologic differences between the murine and human respiratory systems, especially in their upper respiratory tracts (URT). In some models, these differences in the murine nasal cavity can have a significant impact on disease progression and presentation in the lower respiratory tract (LRT) when using intranasal instillation techniques, potentially limiting the usefulness of the mouse model to study these diseases. For these reasons, it would be advantageous to develop a technique to instill bacteria directly into the mouse lungs in order to study LRT disease in the absence of involvement of the URT. We have termed this lung specific delivery technique intubation-mediated intratracheal (IMIT) instillation. This noninvasive technique minimizes the potential for instillation into the bloodstream, which can occur during more invasive traditional surgical intratracheal infection approaches, and limits the possibility of incidental digestive tract delivery. IMIT is a two-step process in which mice are first intubated, with an intermediate step to ensure correct catheter placement into the trachea, followed by insertion of a blunt needle into the catheter to mediate direct delivery of bacteria into the lung. This approach facilitates a &#62;98% efficacy of delivery into the lungs with excellent distribution of reagent throughout the lung. Thus, IMIT represents a novel approach to study LRT disease and therapeutic delivery directly into the lung, improving upon the ability to use mice as surrogates to study human respiratory disease. Furthermore, the accuracy and reproducibility of this delivery system also makes it amenable to Good Laboratory Practice Standards (GLPS), as well as delivery of a wide range of reagents which require high efficiency delivery to the lung.

Intubation-mediated Intratracheal IMIT Instillation: A Noninvasive, Lung-specific Delivery System

Intubation-mediated intratracheal (IMIT) instillation of reagents is an excellent, noninvasive method for studying respiratory disease, as well as a method for instilling therapeutic reagents directly into the lung. It is a rapid and highly reproducible method which is suitable for preclinical testing.

Intubation médiation intratrachéale (GI-TI) instillation: un système de distribution non invasive, Lung-spécifique

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Mouse Pneumonectomy Model of Compensatory Lung Growth

In humans, disrupted repair and remodeling of injured lung contributes to a host of acute and chronic lung disorders which may ultimately lead to disability or death. Injury-based animal models of lung repair and regeneration are limited by injury-specific responses making it difficult to differentiate changes related to the injury response and injury resolution from changes related to lung repair and lung regeneration. However, use of animal models to identify these repair and regeneration signaling pathways is critical to the development of new therapies aimed at improving pulmonary function following lung injury. The mouse pneumonectomy model utilizes compensatory lung growth to isolate those repair and regeneration signals in order to more clearly define mechanisms of alveolar re-septation. Here, we describe our technique for performing mouse pneumonectomy and sham pneumonectomy. This technique may be utilized in conjunction with lineage tracing or other transgenic mouse models to define molecular and cellular mechanism of lung repair and regeneration.

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Method of Isolated Ex Vivo Lung Perfusion in a Rat Model: Lessons Learned from Developing a Rat EVLP Program

Ex-Vivo Lung Perfusion (EVLP) has allowed lung transplantation in humans to become more readily available by enabling the ability to assess organs and expand the donor pool. Here, we describe the development of a rat EVLP program and refinements that allow for a reproducible model for future expansion.

Méthode de isolé<em&gt; Ex Vivo</em&gt; Lung perfusion dans un modèle rat: Leçons tirées élaboration d&#39;un programme EVLP Rat

The number of acceptable donor lungs available for lung transplantation is severely limited due to poor quality. Ex-Vivo Lung Perfusion (EVLP) has allowed ...

A Simple Device to Rapidly Prepare Whole Mounts of the Mouse Intestine

Preparing whole mounts of the mouse small intestine and colon for subsequent analysis or quantification can be time consuming and difficult. We describe the use of a simple device to cut and &#8216;roll&#8217; mouse intestines to rapidly prepare whole mount preparations of superior and uniform quality to that which can be achieved by hand. The device comprises a base that holds 4 stainless steel rods and a top, which acts a cutting guide. The rods are inserted into the lumen of the small intestine [divided into thirds] and the colon. The rods and samples are then placed over a piece of filter paper or card into the holding slots in the base of the device. The top of the device is then positioned and serves as a cutting guide. The two angled sections in the center of the top piece are used to guide a knife or scalpel and cut the intestines longitudinally on the top of the rods. Once the intestinal sections have been cut, the top is removed and the card,&#160;tissue and rods gently removed from the device and placed on the bench. The rods are then gently rolled sideways to flatten and stick the intestinal segments onto the underlying piece of filter paper or card. The final preparation can then be examined or fixed and stored for later analysis. The preparations are invaluable for the study of intestinal changes in normal or genetically modified mouse models. The preparations have been used for the study and quantification of the effects of inflammation (colitis), damage, pre-cancerous lesions (aberrant crypt foci (ACFs) and mucin depleted foci (MDFs)) and polyps or tumors.

The use of a simple device to cut and &#8216;roll&#8217; mouse intestines to rapidly prepare whole mount preparations is described.

Un dispositif simple d&#39;élaborer rapidement entiers Monts de l&#39;intestin de souris

Preparing whole mounts of the mouse small intestine and colon for subsequent analysis or quantification can be time consuming and difficult. We describe ...

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.

This article presents a rapid and simple method for administering bleomycin directly into the mouse trachea via intubation. Key advantages of this method are that it is highly reproducible, easy to master, and does not require specialized equipment or lengthy recovery times.

Une méthode améliorée pour Rapid intubation de la trachée chez la souris

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