이 작품은 NIH K99 HL09222601 및 MNH에게 수여 S &amp; R 재단 Ryaji Uneo 수상에 의해 지원되었다.

1. 마취 <ol><li> isoflurane과 쥐를 Anesthetizing하면 intraperitaneally 마취 관리를하기 전에 동물의 처리와 구속을 촉진 수도 있습니다. 흡입 마취제를 사용하면 또는 특정 연구에 적합하지 않을 수 있습니다 수 있습니다 제안한 폐 연구에서 첫 번째 isoflurane 사용의 영향, 또는 다른 마취제를 결정합니다. </li><li> isoflurane과 동물을 마취. 동영상은 산소와 혼합 2퍼센트 isoflurane에서 설정 isoflurane 기화기 챔버의 사용을 보여줍니다. 당신이 isoflurane 챔버가 없다면, 당신은 표준 또는 오픈 드롭 노출 기법을 사용하실 수 있습니다. 하단에 장착되어 와이어 메쉬와 벨 병 속에 쥐 (또는 다른 적절한 비 다공성 컨테이너)를 동봉. 메쉬 와이어 아래에 위치 거즈 패드하려면, 프로필렌 글리콜에 V / V isoflurane 30 % 혼합물을 적용합니다. 메쉬의 사용은 동물이 피부 자극 및 잠재 도달 했나봐을 일으킬 수있는 isoflurane - 절어 패드와 접촉하지 않도록 보장합니다. isoflurane에 짧은 노출 후, 동물은 anesthetized되어야하며 그릇이 스쳐 및 발가락 핀치 후 응답하지 않습니다 때 righting 반사와 응답하지 않습니다. </li><li> Xylazine과 300ml의 혼합의 내부 복막 주사로 마취 더 마우스. (창자 또는 다른 중요한 장기의 일부가 침투되지 않은 것을 막기 위해 다시 그릴 intraperitoneal 주사를 수행할 때 그것은 좋은 연습입니다.) 0.9 % 멸균 인산의 8850 μL 1000 μL 케타민을 (100 MG / ML) 150 μL를 Xylazine (100 MG / ML) 혼합하면 (PBS) 살린 버퍼. 생쥐의 경우 복용량이 체중의 10 GM 당 약 80 μL 것입니다. 최종 농도는 10 밀리그램 / 킬로그램 Xylazine과 100 밀리그램 / 킬로그램 케타민을 것입니다. 동물 건조 유지 및 체온의 과도한 손실을 방지하기 위해 절연해야합니다. 안과 연고는 각막의 건조 방지하기 위해 눈에 적용할 수 있습니다. </li></ol> 2. 외과 영역의 준비 <ol><li> 수술 절개가 이루어집니다 영역을 면도하지 않은 준비 지역에서 오염을 방지할 수 있습니다. 전기 면도기는 밀접한 동물에서 모피를​​ 면도 있습니다. 면도 지역은 절개 사이트보다 큰해야합니다. </li><li> 같은 나이르로 제모 크림의 후속 응용 프로그램, 모든 남아있는 모피를 제거합니다. 외과 영역에 제모 크림의 두꺼운 레이어를 적용하고 3 분에 둡니다. 젖은 수건으로 모피 및 제모 크림을 닦아. 10 분 초과에서 머리 제거 크림을 두지 마십시오. </li><li> 무균 수술 영역을 준비, 70 % 알코올, 또는 대체 스크럽 솔루션을 사용합니다. </li></ol> 3. 수술 <ol><li> sloped 수술 보드, 또는 직각 접근 금지 서에 anesthetized 동물을 놓습니다. </li><li> 목 앞쪽에 (목 지역) 근처에 작은 절개를합니다. </li><li> tracheal 반지를 시각화하고 액세스하기 위해 platysma과 앞쪽에 tracheal 근육을 멀리 해부하다. </li><li> 일반적으로, intratracheal 사출 볼륨 무게 3 μl 당 그램 (약 30-75 μL 최종 볼륨)입니다. 1mL 피하 (하위 Q) 30 게이지, 5/16-inch 바늘로 주사기를 사용 instillate의 해당 금액을 관리할 수 있습니다. 주사기 베벨 측면을 잡고 병렬 기관과 함께, 그리고기도에 instillate의 전체 볼륨을 주입. </li><li> 동물가 폐 속에 용질의 기지를 따라 온다하여 응답할 수 있습니다. 주사기를 통해 공기를 도입하고 폐가 확장 여부를 관찰하면 성공 삽관법 기법을 확인합니다. </li><li> 기관에서 주사기를 제거합니다. </li></ol> 4. 봉합. <ol><li> 바늘 홀더와 바늘의 스웨이지 (봉합 재료가 연결되어있는 바늘의 두꺼운 부분)을 파악. </li><li> 피부의 두께에 따라 떨어진 상처의 가장자리에서 90 ° 각도로 약 1~3mm에서 바늘의 끝에와 피부에 침투. 조직 하나를 통해 통과 후 종료 바늘을 파악. 개별적으로 매듭 간단한 바늘, 효율적으로 상처를 닫습니다. </li></ol> 5. 동물 복구 <ol><li> 동물 완전히 3-24시간 후 마취 및 수술에서 회복한다. </li></ol> 6. 대표 결과 형광 화합물 Intratracheal 떨어뜨림, dextran을 복합 Cy5.5는 B6 마우스 폐에 형광 화합물 (그림 1) 성공적으로 기지를 보여줍니다. 형광 염료는 주로 폐를 타겟으로하고 균일하게 분포돼있다. 즉시 절차 다음과 같은 위장관의 형광 신호가 식도가 실수 절차 (그림 2) 중 intubated 것을 나타냅니다. 수사 당국은 또한 폐로 화합물의 타겟 배달을 확인하고 instillate의 분포를 확인 폐가 팽창하고 폐 조직 조각 (2, 3) 준비하여 절차를 수행하거나 고정 histolo을gical 샘플. <img src="/files/ftp_upload/1941/1941fig1.jpg" alt="그림 1" /> 그림 1. 형광등 Cy5.5 신호가 성공적으로 얻은 마우스 폐에서 X - 레이 이미지와 함께 공동 등록. 1.4 MG Cy5.5 De​​xtran은 (10 μg / μL의 농도에서) B6 마우스 폐의기도로 얻은되었습니다. Cy5.5 신호는 675 나노미터 여기 필터와 695 nm의 방출 필터를 사용하여 검색했습니다. <img src="/files/ftp_upload/1941/1941fig2.jpg" alt="그림 2" /> 그림 2. 저조한 얻은 마우스 X - 레이 오버레이로 표시 형광 신호의 공동 등록. 내장에 Cy5.5 납품에​​ tracheal 떨어뜨림 프로토콜 결과 동안 식도의 부적 절한 삽관법.

<ol>
	<li>Driscoll, K. E., Costa, D. L., Hatch, G., Henderson, R., Oberdorster, G., Salem, H., Schlesinger, R. B. <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>Helms, M. N., Jain, L., Self, J. L., Eaton, D. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Redox+regulation+of+epithelial+sodium+channels+examined+in+alveolar+type+1+and+2+cells+patch-clamped+in+lung+slice+tissue.">Redox regulation of epithelial sodium channels examined in alveolar type 1 and 2 cells patch-clamped in lung slice tissue.</a> J Biol Chem. 283, 22875-22883 (2008).</li><li>Helms, M. N., Self, J., Bao, H. F., Job, L. C., Jain, L., Eaton, D. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dopamine+activates+amiloride-sensitive+sodium+channels+in+alveolar+type+I+cells+in+lung+slice+preparations.">Dopamine activates amiloride-sensitive sodium channels in alveolar type I cells in lung slice preparations.</a> Am J Physiol Lung Cell Mol Physiol. 291, L610-L618 (2006).</li><li>Iyer, S. S., Ramirez, A. M., Ritzenthaler, J. D., Torres-Gonzalez, E., Roser-Page, S., Mora, A. L., Brigham, K. L., Jones, D. P., Roman, J., 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=Oxidation+of+extracellular+cysteine/cystine+redox+state+in+bleomycin-induced+lung+fibrosis.">Oxidation of extracellular cysteine/cystine redox state in bleomycin-induced lung fibrosis.</a> Am J Physiol Lung Cell Mol Physiol. 296, L37-L45 (2009).</li><li>Li, T., Folkesson, H. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=RNA+interference+for+alpha-ENaC+inhibits+rat+lung+fluid+absorption+in+vivo.">RNA interference for alpha-ENaC inhibits rat lung fluid absorption in vivo.</a> Am J Physiol Lung Cell Mol Physiol. 290, L649-L660 (2006).</li><li>Sager, T., Porter, D., Robinson, V., Lindsley, W., Schwegler-Berry, D., Castranova, V. <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+method+to+disperse+nanoparticles+for+in+vitro+and+in+vivo+investigation+of+toxicity.">An improved method to disperse nanoparticles for in vitro and in vivo investigation of toxicity.</a> Nanotoxicol. 1, 118-129 (2007).</li><li>Xu, J., Gonzalez, E. T., Iyer, S. S., Mac, V., Mora, A. L., Sutliff, R. L., Reed, A., Brigham, K. L., Kelly, 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=Use+of+senescence-accelerated+mouse+model+in+bleomycin-induced+lung+injury+suggests+that+bone+marrow-derived+cells+can+alter+the+outcome+of+lung+injury+in+aged+mice.">Use of senescence-accelerated mouse model in bleomycin-induced lung injury suggests that bone marrow-derived cells can alter the outcome of lung injury in aged mice.</a> J Gerontol A Biol Sci Med Sci. 64, 731-739 (2009).</li><li>Yeh, T. F., Lin, H. C., Chang, C. H., Wu, T. S., Su, B. H., Li, T. C., Pyati, S., Tsai, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+intratracheal+instillation+of+budesonide+using+surfactant+as+a+vehicle+to+prevent+chronic+lung+disease+in+preterm+infants:+a+pilot+study.">Early intratracheal instillation of budesonide using surfactant as a vehicle to prevent chronic lung disease in preterm infants: a pilot study.</a> Pediatrics. 121, e1310-e1318 (2008).</li></ol>

Intratracheal instillations는 시험 화합물 (6)의 독성을 평가하기 위해 여러 가지 다양한 연구에 사용되었으며, 폐포 폐 상해 (4, 7) 유​​도, 직접 작은 oligonucleotides의 전달을 통해 계면 활성제 (8)뿐​​만 아니라, 변경 유전자 발현을 교체 폐 (5). 우리는 현재 폐암에 형광 산화 환원에 민감한 자료는 약리 성분, DNA 벡터, 바이러스, 그리고 함께 형광 표시기를 instilling)뿐만 아니라 2로 평범하고 pathophysiolog...

<table><tbody><tr><td>Isoflurane</td><td>Webster Veterinary</td><td>14043-220-05</td><td></td></tr><tr><td>Xylazine</td><td>Lloyd, Inc.</td><td></td><td>New Animal Drug Application #139-236</td></tr><tr><td>Ketamine</td><td>Bioniche Pharma</td><td>67457-001-10</td><td></td></tr><tr><td>Nair</td><td></td><td></td><td>Available at drug stores</td></tr><tr><td>1mL SubQ Syringe</td><td>BD Biosciences</td><td>309597</td><td>26 5/8 G</td></tr><tr><td>4-0 Nylon Suture</td><td>Ethicon Inc.</td><td>1894G.S30</td><td></td></tr></tbody></table>

direct tracheal instillation of solutes into mouse lung

Intratracheal instillations 직접 폐에 solutes을 제공합니다. 이 절차는 폐암의 말초 지역에 instillate의 전달을 대상으로하고, 따라서 종종 폐포를 공부하기위한 연구에 통합됩니다. 우리는 생쥐에서 intratracheal instillations을 수행하기위한 세부적인 생존 프로토콜을 제공합니다. 이 방법을 사용, 하나는 말초 폐로 테스트 solutes 또는 고체의 제공 (예 : 폐 치료제, 계면 활성제, 바이러스, 작은 oligonucleotides)를 타겟팅할 수 있습니다. Tracheal instillations는 주로 상부 호흡기를 타겟으로하고 가능한 잠재적 유해 물질에 대한 조사에 노출될 수 있습니다 흡입 프로토콜을 통해, 원하는 방법 수 있습니다. 또한, tracheal 떨어뜨림 프로토콜을 사용하여, 동물 완전히 비침습 절차에서 복구할 수 있습니다. 이 같은 동물을 사용하여 시험 동물, 또는 다시 설치에 대한 후속 생리 측정을 만들기위한 수 있습니다. 폐에 소개 instillate의 금액은 신중하게 결정과 동물 복구를 보장하기 위해 osmotically 균형해야합니다. 일반적으로 30-75 μL instillate 볼륨 마우스 폐에 소개하실 수 있습니다.

Watch this Scientific Journal Video about Direct Tracheal Instillation of Solutes into Mouse Lung at JoVE.com

Direct Tracheal Instillation of Solutes into Mouse Lung

Intratracheal instillations 직접 폐에 solutes을 제공합니다. 이 절차는 폐암의 말초 지역에 instillate의 전달을 대상으로하고, 따라서 종종 폐포를 공부하기위한 연구에 통합됩니다. 우리는 생쥐에서 intratracheal instillations을 수행하기위한 세부적인 생존 프로토콜을 제공합니다.

마우스 폐에 Solutes 직접 Tracheal 떨어뜨림

Intratracheal instillations deliver solutes directly into the lungs.  This procedure targets the delivery of the instillate into the distal regions of the ...

direct-tracheal-instillation-of-solutes-into-mouse-lung

Research

JoVE Journal

Biology

29.0K 조회수.  Emory University. Intratracheal instillations 직접 폐에 solutes을 제공합니다. 이 절차는 폐암의 말초 지역에 instillate의 전달을 대상으로하고, 따라서 종종 폐포를 공부하기위한 연구에 통합됩니다. 우리는 생쥐에서 intratracheal instillations을 수행하기위한 세부적인 생존 프로토콜을 제공합니다.

비디오: Direct Tracheal Instillation of Solutes into Mouse Lung

Non-surgical Intratracheal Instillation of Mice with Analysis of Lungs and Lung Draining Lymph Nodes by Flow Cytometry

Phagocytic cells such as alveolar macrophages and lung dendritic cells (LDCs) continuously sample antigens from the alveolar spaces in the 
lungs. LDCs, in particular, are known to migrate to the lung draining lymph nodes (LDLNs) where they present inhaled antigens to T cells initiating an 
appropriate immune response to a variety of immunogens1,2. To model interactions between the lungs and airborne antigens in mice, antigens can be 
administered intranasally1,3,4, intratracheally5 or as aerosols6. Delivery by each route involves distinct technical skills and limitations that need to be 
considered before designing an experiment. For example, intranasal and aerosolized exposure delivers antigens to both the lungs and the upper respiratory 
tract. Hence antigens can access the nasal associated lymphoid tissue (NALT)7, potentially complicating interpretation of the results. In addition, swallowing, 
sneezing and the breathing rate of the mouse may also lead to inconsistencies in the doses delivered. Although the involvement of the upper respiratory tract may 
be preferred for some studies, it can complicate experiments focusing on events specifically initiated in the lungs. In this setting, the intratracheal (i.t) 
route is preferable as it delivers test materials directly into the lungs and bypasses the NALT. Many i.t injection protocols involve either blind intubation of the 
trachea through the oral cavity or surgical exposure of the trachea to access the lungs. Herein, we describe a simple, consistent, non-surgical method for i.t 
instillation. The opening of the trachea is visualized using a laryngoscope and a bent gavage needle is then inserted directly into the trachea to deliver the 
innoculum. We also describe procedures for harvesting and processing of LDLNs and lungs for analysis of antigen trafficking by flow cytometry.

We illustrate non-surgical delivery of test materials into the lungs of anesthetized mice via the trachea. This method permits lung exposure to bacterial and viral pathogens, cytokines, antibodies, beads, chemicals, or dyes. We further describe harvesting and processing of lungs and lung draining lymph nodes (LDLNs) for flow cytometry.

유동세포계측법에 의해 폐, 폐 배수 림프절의 분석과 마우스의 비 수술 Intratracheal 떨어뜨림

Phagocytic cells such as alveolar macrophages and lung dendritic cells (LDCs) continuously sample antigens from the alveolar spaces in the 
lungs. LDCs, ...

연구

면역학 및 감염병학

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.

마우스의 기관 내 삽관<em&gt;를 통해</em이경 사용&gt; 직접 후두경

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

의학

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.

마우스 폐 삽관 법의 간단한 방법

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

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.

비 침습적 기관 내 삽관 마우스 폐의 병리학 및 생리학을 연구하는

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 ...

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.

삽관 매개 기관 내 (IMIT) 떨어 뜨림 : 비 침습적, 폐 별 전달 시스템

Respiratory disease studies typically involve the use of murine models as surrogate systems. However, there are significant physiologic differences ...

Instillation and Fixation Methods Useful in Mouse Lung Cancer Research

The ability to instill live agents, cells, or chemicals directly into the lung without injuring or killing the mice is an important tool in lung cancer research. Although there are a number of methods that have been published showing how to intubate mice for pulmonary function measurements, none are without potential problems for rapid tracheal instillation in large cohorts of mice. In the present paper, a simple and quick method is described that enables an investigator to carry out such instillations in an efficient manner. The method does not require any special tools or lighting and can be learned with very little practice. It involves anesthetizing a mouse, making a small incision in the neck to visualize the trachea, and then inserting an intravenous catheter directly. The small incision is quickly closed with tissue adhesive, and the mice are allowed to recover. A skilled student or technician can do instillations at an average rate of 2 min/mouse. Once the cancer is established, there is frequently a need for quantitative histologic analysis of the lungs. Traditionally pathologists usually do not bother to standardize lung inflation during fixation, and analyses are often based on a scoring system that can be quite subjective. While this may sometime be sufficiently adequate for gross estimates of the size of a lung tumor, any proper stereological quantification of lung structure or cells requires a reproducible fixation procedure and subsequent lung volume measurement. Here we describe simple reliable procedures for both fixing the lungs under pressure and then accurately measuring the fixed lung volume. The only requirement is a laboratory balance that is accurate over a range of 1 mg&#8211;300 g. The procedures presented here thus could greatly improve the ability to create, treat, and analyze lung cancers in mice.

The goal of this paper is to describe simple methods that will greatly aid in the setup and analysis of mouse lungs with lung cancer or other pathologies. We present 3 protocols to simply and reliably carry out lung instillations, fixation, and lung volume measurements.

마우스 폐 암 연구에 유용한 점안 및 고정 방법

The ability to instill live agents, cells, or chemicals directly into the lung without injuring or killing the mice is an important tool in lung cancer ...

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.

마우스의기도 신속 삽관 용 개선 된 방법

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

Intratracheal Administration of Dry Powder Formulation in Mice

In the development of inhalable dry powder formulations, it is essential to evaluate their biological activities in preclinical animal models. This paper introduces a noninvasive method of intratracheal delivery of dry powder formulation in mice. A dry powder loading device that consists of a 200 &#181;L gel loading pipette tip connected to an 1 mL syringe via a three-way stopcock is presented. A small amount of dry powder (1-2 mg) is loaded into the pipette tip and dispersed by 0.6 mL of air in the syringe. Because pipette tips are disposable and inexpensive, different dry powder formulations can be loaded into different tips in advance. Various formulations can be evaluated in the same animal experiment without device cleaning and dose refilling, thereby saving time and eliminating the risk of cross-contamination from residual powder. The extent of powder dispersion can be inspected by the amount of powder remaining in the pipette tip. A protocol of intubation in mouse with a custom-made light source and a guiding cannula is included. Proper intubation is one of the key factors that influences the intratracheal delivery of dry powder formulation to the deep lung region of the mouse.

Dry powder formulations for inhalation have great potential in treating respiratory diseases. Before entering human studies, it is necessary to evaluate the efficacy of the dry powder formulation in preclinical studies. A simple and noninvasive method of the administration of dry powder in mice through the intratracheal route is presented.

쥐에서 건조 분말 제형의 장내 투여

In the development of inhalable dry powder formulations, it is essential to evaluate their biological activities in preclinical animal models. This paper ...

A Minimally Invasive Method for Intratracheal Instillation of Drugs in Neonatal Rodents to Treat Lung Disease

Treatment of neonatal rodent with drugs instilled directly into the trachea could serve as a valuable tool to study the impact of a locally administered drug. This has direct translational impact because surfactant and drugs are administered locally into the lungs. Though the literature has many publications describing minimally invasive transoral intubation of adult mice and rats in therapeutic experiments, this approach in neonatal rat pups is lacking. The small size of orotracheal region/pharynx in the pups makes visualization of laryngeal lumen (vocal cords) difficult, contributing to the variable success rate of intratracheal drug delivery. We hereby demonstrate effective oral intubation of neonatal rat pup - a technique that is non-traumatic and minimally-invasive, so that it can be used for serial administration of drugs. We used an operating otoscope with an illumination system and a magnifying lens to visualize the tracheal opening of the rat neonates. The drug is then instilled using a 1 mL syringe connected to a pipette tip. The accuracy of the delivery method was demonstrated using Evans blue dye administration. This method is easy to get trained in and could serve as an effective way to instill drugs into trachea. This method could also be used for administration of inoculum or agents to simulate disease conditions in animals and, also, for cell-based treatment strategies for various lung diseases.

This technique of instilling drugs directly into the trachea of neonatal rodents is important in studying the impact of locally administered drugs or biologicals on neonatal lung diseases. Additionally, this method can also be used for inducing lung injury in animal models.

Treatment of neonatal rodent with drugs instilled directly into the trachea could serve as a valuable tool to study the impact of a locally administered ...

Noninvasive Intratracheal Lipopolysaccharide Instillation in Mice

The acute lung injury (ALI) mouse model induced by lipopolysaccharide (LPS) or endotoxin is still among the most commonly used models in animal studies of acute lung injury or acute inflammation. The current most commonly used methods in acute lung injury mouse models are an intraperitoneal injection of LPS and tracheostomy for the tracheal infusion of LPS. However, the former method lacks lung targeting and damages other organs, and the latter method induces operative trauma, infection risk, and a low survival rate. Here, we recommend a noninvasive oropharyngeal endotracheal intubation method for LPS instillation in mice. In this method, LPS is noninvasively introduced into the trachea through the oropharyngeal cavity to be instilled into the lung with the help of an apparatus for endotracheal intubation. This method not only assures lung targeting but also avoids damage and the risk of death in the animals. We expect that this approach will become widely used in the field of acute lung injury.

Here, we propose a protocol for intratracheal lipopolysaccharide (LPS) delivery via noninvasive oropharyngeal endotracheal intubation. This method minimizes the trauma of the surgical procedure for the animal and accurately delivers LPS to the trachea and then to the lungs.

The acute lung injury (ALI) mouse model induced by lipopolysaccharide (LPS) or endotoxin is still among the most commonly used models in animal studies of ...