Name: repeated orotracheal intubation in mice
Uploaded: 2020-03-27T14:00:00
Description: Watch this Scientific Journal Video about Repeated Orotracheal Intubation in Mice at JoVE.com

作者感谢露西娅·罗莎斯、劳伦·杜利特尔、丽莎·约瑟夫和林赛·弗格森的技术援助，感谢大学实验室动物资源组织对动物保护的支持。这项工作由NIH T35OD010977和R01-HL102469资助。

此处描述的所有动物活动均已获得俄亥俄州立大学机构动物护理和使用委员会 （IACUC） 的批准，并在 AAALAC 认可的设施中进行。
1. 程序准备
<ol><li>构建插管平台。要达到适当的平台斜率，请使用三英寸（7.6 厘米）3 环活页夹。将 15⁄20 厘米长的 3-0 丝或其他螺纹材料折成两半，用胶带将螺纹的末端粘附到倾斜平台的顶部，以创建悬架环（图 1）。</li>
	<...

<ol>
	<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 (1), 128-135 (2007).</li><li>Rivera, B., Miller, S. R., Brown, E. M., Price, R. E. <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. 44 (2), 52-55 (2005).</li><li>Sparrowe, J., Jimenez, M., Rullas, J., Martinez, A. E., Ferrer, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Refined+Intratracheal+Intubation+Technique+in+the+Mouse,+Complete+Protocol+Description+for+Lower+Airway+Models.">Refined Intratracheal Intubation Technique in the Mouse, Complete Protocol Description for Lower Airway Models.</a> Global Journal of Animal Scientific Research. 3 (2), 363-369 (2015).</li><li>Deyo, D. J., Wei, J. <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+of+Intubation+and+Ventilation+in+Mice.">A Novel Method of Intubation and Ventilation in Mice.</a> Anesthesia &amp; Analgesia. 88 (2), 179 (1999).</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 (3), 103-106 (2004).</li><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> Journal of Applied Physiology. 87 (6), 2362-2365 (1999).</li><li>Das, S., MacDonald, K., Chang, H. 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+Simple+Method+of+Mouse+Lung+Intubation.">A Simple Method of Mouse Lung Intubation.</a> Journal of Visualized Experiments. (73), e50318 (2013).</li><li>Limjunyawong, N., Mock, J., 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=Instillation+and+Fixation+Methods+Useful+in+Mouse+Lung+Cancer+Research.">Instillation and Fixation Methods Useful in Mouse Lung Cancer Research.</a> Journal of Visualized Experiments. (102), e52964 (2015).</li><li>Qin, W., Baran, U., Wang, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lymphatic+response+to+depilation-induced+inflammation+assessed+with+label-free+optical+lymphangiography.">Lymphatic response to depilation-induced inflammation assessed with label-free optical lymphangiography.</a> Lasers in Surgery and Medicine. 47 (8), 669-676 (2015).</li><li>McGovern, T. K., Robichaud, A., Fereydoonzad, L., Schuessler, T. F., Martin, J. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+Respiratory+System+Mechanics+in+Mice+using+the+Forced+Oscillation+Technique.">Evaluation of Respiratory System Mechanics in Mice using the Forced Oscillation Technique.</a> Journal of Visualized Experiments. (75), e50107 (2013).</li></ol>

采用横皮气管可视化技术的插管技术为标准皮肤切口方法提供了一种精细的方法。特别注意几个关键步骤，可以轻松快速地实现插管。动物必须被放置在插管平台上的背底中，用轻柔的回缩固定。这将延长动物到垂直对齐和适当的定位插管。此外，脱毛霜不应与动物的皮肤接触超过30~45s，并应彻底冲洗以清除所有残留物。延长皮肤接触脱毛霜会造成不必要的疼痛的动物和溃疡可以阻碍气管<sup class...

实验鼠是人类呼吸道疾病的常见动物模型。因此，有几种已公布的小鼠插管方法，用于灌输治疗和测量呼吸力学。大部分所述程序要求透过口腔对牙洞进行化形，并配备专用设备，例如喉镜或光纤光源121、2、3、4、5、6、7。5,6,73,4,,,然而，当需要相对较大的管条时，这可能很困难，因为它会模糊研究人员的观点。Limjunyawong等人8日用插管的方法解决了这个问题，在插管方法中，沿着腹腔颈的中线进行小切口，以便对气管进行可视化。手术后，切口用组织胶粘剂封闭。
对于需要频繁重复插管的研究，该部位的连续切口和关闭需要使皮肤边缘和腹腔颈的组织创伤被除污。口服插管的跨皮气管可视化方法的目的是提供一种精细的非侵入性技术，专门适用于小鼠的重复插管研究以及单插管事件。

<table>
	<tbody>
		<tr>
			<td>18Gx1 1/4&#34; intravenous catheter, Safelet</td>
			<td>Fisher Scientific</td>
			<td>#14-841-14</td>
			<td>Cannula for intubation</td>
		</tr>
		<tr>
			<td>70% ethanol, 10L</td>
			<td>Fisher Scientific</td>
			<td>25467025</td>
			<td>Cleaning cannula</td>
		</tr>
		<tr>
			<td>Abrasive paper (sandpaper)</td>
			<td>Porter-Cable</td>
			<td>74001201</td>
			<td>Cannula preparation</td>
		</tr>
		<tr>
			<td>AnaSed (xylazine sterile solution) injection (100 mg/ml)</td>
			<td>Akorn Animal Health</td>
			<td>NDC# 59399-111-50</td>
			<td>Anesthesia</td>
		</tr>
		<tr>
			<td>Blue labeling tape (0.5 in x 14 yds)</td>
			<td>Fisher Scientific</td>
			<td>15966</td>
			<td>Restraint on intubation platform</td>
		</tr>
		<tr>
			<td>Braided silk suture without needle, nonsterile, (3-0)</td>
			<td>Henry Schein</td>
			<td>Item #1007842</td>
			<td>Intubation platform</td>
		</tr>
		<tr>
			<td>Deltaphase Isothermal Pad</td>
			<td>Braintree Scientific</td>
			<td>39DP</td>
			<td>Mouse thermoregulation and recovery</td>
		</tr>
		<tr>
			<td>Deltaphase Operating Board</td>
			<td>Braintree Scientific</td>
			<td>39OP</td>
			<td>Mouse recovery (prior to extubation)</td>
		</tr>
		<tr>
			<td>Distilled water</td>
			<td>ThermoFisher</td>
			<td>15230253</td>
			<td>Cleaning mouse following depilation</td>
		</tr>
		<tr>
			<td>Eye Scissors, angled, sharp/sharp</td>
			<td>Harvard Apparatus</td>
			<td>72-8437</td>
			<td>Cannula preparation</td>
		</tr>
		<tr>
			<td>FlexiVent (FX Module 2)</td>
			<td>Scireq</td>
			<td>N/A</td>
			<td>Record lung function data (not required to perform procedure, used in this study to validate procedure)</td>
		</tr>
		<tr>
			<td>Gauze sponges</td>
			<td>Fisher scientific</td>
			<td>13-761-52</td>
			<td>Hair removal</td>
		</tr>
		<tr>
			<td>Heavy-Duty 3&#34; 3-Ring View Binders</td>
			<td>Staples</td>
			<td>24690CT</td>
			<td>Intubation platform</td>
		</tr>
		<tr>
			<td>Instat Software</td>
			<td>Graphpad</td>
			<td>N/A</td>
			<td>Statistical analysis software</td>
		</tr>
		<tr>
			<td>Insulin syringe (0.5 cc, U100)</td>
			<td>Fisher Scientific</td>
			<td>329461</td>
			<td>Anesthesia administration</td>
		</tr>
		<tr>
			<td>Ketamine HCl Injection, USP (100 mg/ml)</td>
			<td>Llyod Laoratories</td>
			<td>List No. 4871</td>
			<td>Anesthesia</td>
		</tr>
		<tr>
			<td>Lung inflation bulb</td>
			<td>Harvard Apparatus</td>
			<td>72-9083</td>
			<td>Confirm cannula placement</td>
		</tr>
		<tr>
			<td>Micro Forceps, Curved, Smooth</td>
			<td>Harvard Apparatus</td>
			<td>72-0445</td>
			<td>Retract tongue and create tension on neck for cannula visualization</td>
		</tr>
		<tr>
			<td>Nair (hair removal lotion), 9 oz bottle</td>
			<td>Church &#38; Dwight</td>
			<td>42010440</td>
			<td>Hair removal</td>
		</tr>
		<tr>
			<td>Sterile saline (0.9%), 10 ml</td>
			<td>Fisher Scientific</td>
			<td>NC9054335</td>
			<td>Anesthesia, cleaning skin following hair removal</td>
		</tr>
	</tbody>
</table>

repeated orotracheal intubation in mice

文献描述了几种小鼠插管方法，这些方法要么需要通过口腔对glottis进行可视化，要么要求在腹腔颈部进行切口的可视化，以便直接确认气管中的切口位置。此类程序对受试者引起的相对困难或组织创伤可能妨碍调查员进行纵向研究的能力。本文演示了一种技术，即使用脱毛器从腹腔颈部去除毛发后对小鼠进行物理操作，无论皮肤程度如何，气管的截皮可视化都可用于排泄管。色素 沉着。这种方法对主体无害，对鼠解剖结构的理解有限，很容易实现。这种精细的方法有助于反复插管，这可能是监测疾病进展或灌输治疗的所必需的。使用这种方法可能会减少在呼吸疾病小鼠模型中测量肺功能所需的动物数量和技术技能。

基线肺功能的串行监测 18周大的雌性BALB/c和10周大的C57BL/6小鼠（每株中的3只）在第0天、3日、10日和17日使用所述方法插管。每天插管后，受试者连接到一个机械呼吸机，提供100%氧气（材料表）。呼吸阻力 （Rrs） 在深膨胀到 25 厘米 H2O 保持 5 s 后，使用强制振荡技术测量了 60 s。与这种持续呼吸保持和生理范围内的Rrs值相关的软件错误为正确放置导管提供了额?...

Watch this Scientific Journal Video about Repeated Orotracheal Intubation in Mice at JoVE.com

Repeated Orotracheal Intubation in Mice

本文的目的是描述实验室小鼠插管的精细方法。该方法是非侵入性的，因此非常适合需要对呼吸功能进行连续监测和/或将治疗灌输到肺部的研究。

小鼠重复的奥内切管插管

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

该协议将允许研究人员对单个小鼠的肺功能进行重复测量，减少实验变异性，减少每个实验所需的小鼠数量。该协议的主要优点是，它不需要通过口腔或颈部的创伤性解剖可视化插管。这种方法对于研究慢性和渐进性肺病（如哮喘、慢性阻塞性肺病、肺气肿和肺纤维化）的研究人员具有特别的价值。该协议的视觉演示至关重要，因为了解如何通过腹颈皮肤识别气管是方法的核心。与安德鲁·纳尔逊一起展示这项技术的将是凯特·诺兰，一位来自我实验室的兽医。要创建插管平台，请将长度 15 到 20 厘米的 3-0 丝折成两半，并将螺纹的末端绑在三英寸三环粘结器的顶部。选择适当尺寸和长度的导管，并在导管的端角切一个斜角。用研磨纸平滑切割表面，形成圆角，并在离斜面约一厘米的弯管中轻轻弯曲。当材料准备就绪时，确认麻醉鼠标对手趾捏没有反应，并在平台倾斜表面顶部的丝丝螺纹周围钩上切口，将鼠标悬挂在上部位置。一旦鼠标被方形地定位在背的直卧中，轻轻抓住尾巴的底座，然后向桌子缩回尾巴。将一块胶带放在尾巴底部，以固定鼠标，并在腹颈区域涂抹脱毛霜。30 至 45 秒后，使用干纱布去除奶油，并用盐水或蒸馏水彻底冲洗皮肤。在非显性手中使用钳子，轻轻缩回舌头，以充分张开嘴引入管。用占统治地位的手，推进管伸入嘴里，使是一个显着的端到轻微的弯曲是反对动物的嘴的屋顶。松开舌头，沿着心颈滑动闭合钳的平边，直到达到毛骨膜，横向置换唾液腺，并压平覆盖气管的肌肉。气管将以白线作为皮经出现。如有必要，以颅骨方向旋转钳子，同时保持皮肤上的张力，使横向置换的唾液腺达到峰值，并在气管周围产生更多的对比度。用手腕的同步弯曲来提升占统治地位的手，推进管，同时使管的侧尖心尖。如果管管已正确放置，将在气管内观察。要确认正确放置，请将肺膨胀球与管状图连接，并观察与装置并发凹陷的胸腔扩张。在不更换管管的情况下，小心地将鼠标的切口从插管平台上拔下，将鼠标转移到水平平台上，然后将插拔器连接到呼吸机上的适配器上，并在深度膨胀后，在测量呼吸阻力之前将鼠标通风 60 秒。一旦手术完成，将鼠标转移到加热平台，通过光头或尾捏提供持续刺激，以鼓励自发呼吸。一旦观察到自发呼吸，继续目视监测动物。当呼吸速率恢复正常时，抓住轮毂水平的导管，轻轻将管颅和远离小鼠，直到从动物的嘴里完全去除导管。一旦被挤出，将鼠标转移到一个干净的恢复笼与热支持与监测，直到完全流动。在这个具有代表性的实验中，在第0天、第3天、第10天和17日测量了呼吸阻力，使用强制振荡技术，在深度膨胀到25厘米的水后，保持5秒和60秒的机械通风。每个菌株中的任何时间点之间没有观察到测量的呼吸阻力有显著差异，这表明呼吸道系统连续四个时间点缺乏功能性显著的插管性创伤相关炎症。这种插管程序可用于提供气管的安装实验物质和呼吸功能的序列监测。该技术允许评估肺功能，无需侵入性气管切除术，便于在同一小鼠内慢性和渐进性疾病的多个时间点的监测。

Research

JoVE Journal

Medicine

A Novel Rescue Technique for Difficult Intubation and Difficult Ventilation

一个新型插管困难和困难通风救援技术

We describe a novel non surgical technique to maintain oxygenation and ventilation in a case of difficult intubation and difficult ventilation, which ...

科研

医学

Transthoracic Echocardiography in Mice

经胸超声心动图在小鼠

In recent years, murine models have become the primary avenue for studying the molecular mechanisms of cardiac dysfunction resulting from changes in gene ...

Guidelines for Elective Pediatric Fiberoptic Intubation

选修儿科光纤插管指南

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

Basic Surgical Techniques in the G&ouml;ttingen Minipig: Intubation, Bladder Catheterization, Femoral Vessel Catheterization, and Transcardial Perfusion

Basic Surgical Techniques in the GÃƒÆ’Ã‚Â¶ttingen Minipig: Intubation, Bladder Catheterization, Femoral Vessel Catheterization, and Transcardial Perfusion

基本外科技术，在哥廷根微型猪：气管插管，膀胱导尿，股血管的导管和Transcardial灌注

The emergence of the G&ouml;ttingen minipig in research of topics such as neuroscience, toxicology, diabetes, obesity, and experimental surgery reflects ...

Endotracheal Intubation in Mice via Direct Laryngoscopy Using an Otoscope

Endotracheal Intubation in Mice via Direct Laryngoscopy Using an Otoscope

气管插管小鼠<em&gt;通过</em&gt;直接喉镜使用耳镜

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

一个简单的方法小鼠肺插管

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

无创性气管内插管，以研究小鼠肺的病理生理

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 ...

Carotid Artery Infusions for Pharmacokinetic and Pharmacodynamic Analysis of Taxanes in Mice

颈动脉输液的紫杉烷类化合物在小鼠体内的药代动力学和药效学分析

When proposing the use of a drug, drug combination, or drug delivery into a novel system, one must assess the pharmacokinetics of the drug in the study ...

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

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

插管介导气管内（IMIT）灌输：一种无创，龙专用输送系统

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

An Improved Method for Rapid Intubation of the Trachea in Mice

在小鼠气管插管快速的改进方法

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