Name: use of an integrated low-flow anesthetic vaporizer, ventilator, and physiological monitoring system for rodents
Uploaded: 2020-07-09T13:00:00.000+00:00
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所有动物研究都得到了普渡动物护理和使用委员会的批准。
1. 低流量蒸发器的设置
<ol><li>伊索夫卢兰或塞沃夫鲁兰交付<ol>
<li>选择载体气源。要利用内部空气泵，请从系统背面的入口端口取出红色盖，使系统能够接收室内空气。要使用压缩气体，请使用设置为 15 PSI 的压力调节器或减压器，并连接到系统背面的压缩气体端口。</li>
		<li>将木炭罐连接到排气口。</li>
		<li>将附件连接器连接到系统前面的激励和过期端口。将感应室连接到带有蓝色夹子的树枝和鼻锥连接到带有白色夹子的树枝（图 1）。</li>
	</ol>
</li>
	<li>用于机械通风<ol>
<li>将连接连接器管连接到黄色编码剪辑（图 2）。</li>
		<li>通过执行死空间校准来校准呼吸机。从 通风运行屏幕，触摸 设置，然后 校准和测试。选择 死空间校准 并按 拨号 B。</li>
	</ol>
</li>
	<li>用于脉冲氧化<ol>
<li>将传感器连接到系统背面的端口，标记为 MouseSTAT。</li>
	</ol>
</li>
	<li>用于变暖<ol>
<li>将加热垫连接到系统前面的"垫电源"端口。</li>
		<li>将一个传感器连接到"身体传感器"端口，另一个传感器连接到"垫传感器"端口。将垫传感器固定到加热垫上。</li>
	</ol>
</li>
</ol>2. 配置设置
<ol><li>麻醉<ol>
<li>麻醉系统电源。从 阿内斯特运行屏幕，触摸 设置。</li>
		<li>选择麻醉剂。触摸 类型阿内斯特，然后转动拨号B选择 伊索夫卢兰 或 塞沃夫鲁兰。</li>
		<li>设置注射器大小。触摸 注射器大小，然后转动拨号B以选择大小。</li>
		<li>触摸返回返回到 阿内斯特运行屏幕。</li>
		<li>使用瓶顶适配器，用麻醉剂填充注射器。</li>
		<li>将注射器连接到麻醉系统。如果需要，触摸 删除 以向后移动推手块。</li>
		<li>将注射器主热。触摸并保持 Prime 向前移动推手块，直到推手块接触注射器柱塞的顶部。按住 Prime 按钮时转动 B 会调节推手块速度。</li>
	</ol>
</li>
	<li>用于机械通风<ol>
<li>触摸 文特运行屏幕 选项卡，然后 设置。</li>
		<li>触摸 体重 并输入动物的重量。</li>
		<li>触摸 优先选择 音量或压力控制通风。体重设置自动设置适当的呼吸速率和潮汐体积。</li>
	</ol>
</li>
	<li>用于脉冲氧化<ol>
<li>触摸 Oxi 运行屏幕 选项卡，然后 设置。</li>
		<li>触摸 HR 并转动拨号 B 以设置允许的最低心率读数。预设可用。</li>
	</ol>
</li>
	<li>用于变暖<ol>
<li>从 暖运行屏幕，触摸 设置。选择加热方法和目标温度设置。</li>
	</ol>
</li>
</ol>3. 开始麻醉分娩
<ol><li>麻醉鼠标<ol>
<li>从 Anest 运行屏幕，触摸 启动感应 开始气流。默认感应流速为 500 mL/min。转动拨号 A 可根据需要调整流速。</li>
		<li>将鼠标放在感应室中，紧紧地合上盖子。将 麻醉剂浓度 表盘调整为 3% 的异氟化剂。</li>
		<li>监测，直到小鼠到达所需的麻醉平面，由呼吸速率下降和右反射丧失时，室倾斜。根据需要调整 麻醉剂浓度 表盘。</li>
		<li>一旦动物失去了右反射，并充分麻醉，触摸 停止感应。</li>
		<li>如果需要，触摸 冲洗室 以清空残留麻醉气体的腔室。</li>
		<li>打开通往鼻锥的夹子，并关闭通向腔室的夹子。</li>
		<li>触摸 启动鼻锥。体重设置确定鼻锥流速率，但可以通过转动拨号 A 手动调整。</li>
		<li>立即安装鼻锥，并将动物放在红外加热垫上。</li>
		<li>将动物传感器插入作为直肠探头。</li>
	</ol>
</li>
</ol>4. 开始机械通风
<ol><li>灌管动物。<ol>
<li>将动物转移到插管阶段，同时保持动物麻醉。</li>
		<li>使用固定在垂直插管阶段的螺纹（图3）将动物从上切口吊起。</li>
		<li>轻轻地将动物的舌头放在一边，使用灌装套件中提供的灯光可视化气管。</li>
		<li>小心地插入气管，通过将小气囊连接到管子并检查肺部是否膨胀来验证正确位置。</li>
	</ol>
</li>
	<li>将内脏管连接到通风管。</li>
	<li>触摸 停止鼻锥，然后触摸 启动呼吸机。 注意:体重设置自动确定适当的呼吸速率和潮汐量。要执行压力控制通风，将目标吸入压力设置在 15-18 厘米 H2O 之间。根据手术方案需要对呼吸机设置进行调整。</li>
</ol>5. 开始生理监测
<ol><li>将传感器放在动物的后爪上（图4）。脉冲氧化计将自动开始读取HR和SPO2。 触摸 Oxi 运行屏幕选项卡 以查看脉冲氧化数据。</li>
</ol>

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这个项目得到了肯特科学公司的设备和资金的支持。作者克里斯塔·比贾雷利和戴夫·菲茨米勒是肯特科学公司的员工，该公司生产本文中使用的设备。本文的开放访问出版物由肯特科学公司赞助。

这种数字低流量麻醉系统将麻醉、通风、发热和生理监测系统集成到单个设备中。此外，该系统还包含一个内部泵，允许它吸收环境空气作为载体气体，从而无需压缩气体来源。
在此过程中，该系统被用作唯一的设备，以取代麻醉蒸发器、机械呼吸机、脉冲氧化计和加热垫。我们之前演示过以100mL/min2的流速进行麻醉分娩。流速设置对于这种麻醉输送技术至关重要，因为流速直接控制使用的液体麻醉剂的体积。我们以前也演示过如何使用低流量来节省麻醉液1，2。当传统的蒸发器连接到机械呼吸机时，蒸发器必须连续运行，而呼吸机从气流中抽取样品。在带集成呼吸机的数字蒸发器中，只有通风机输出的通风所需的气体。这降低了与麻醉液、载体气体和木炭过滤器相关的成本。
虽然使用低流量数字蒸发器有很多优点，但也有局限性。该系统设计为以低流速运行，非常适合啮齿动物和其他小型哺乳动物，但不会提供高于1000 mL/min流量的麻醉。因此，这种特殊的系统只适合小型动物物种。集成脉冲氧化计仅包括用于爪子的传感器。不建议在尾部使用传感器，这可能是某些外科手术的限制。此外，虽然可以通过这个系统通过爪子传感器监测呼吸速率，但很难在较长时间内获得一致的呼吸记录。最后，与传统的蒸发器不同，这种数字系统需要电力。电池可用于无法供电或停电的情况下，并且可以通过几个小时的使用为系统供电。
此设置和协议证明安全有效地使用数字低流量麻醉系统与集成呼吸机和生理监测模块。这种设置对于任何长凳空间有限的实验室或在手术场附近存放多件设备和管子不可行的地方都是有用的。一体式系统有许多好处，包括消除压缩气箱和单独的生理监测设备。总体而言，使用传统蒸发器不理想的团体可以考虑这一集成系统。

涉及动物模型的研究往往需要专门的数据收集设备。有两种常见的麻醉蒸发器，通常用于小型动物手术。传统的麻醉蒸发器依靠基于大气压力和气体流动1，2，3，4，5，6，7，8，9，10的挥发性麻醉剂的被动蒸发。它们设计为以0.5升/分钟至10升/分钟的流速运行，是大型动物模型11的理想之选。
与传统的蒸发器12、13相比，我们最近展示了低流量数字蒸发器的效果。低流量的数字麻醉系统可用于保持动物在鼻锥上的极低流速1.5-2.2倍的动物的分钟体积14，15，16。
使用数字麻醉系统有许多好处。它集成了一个内置泵，它吸收环境空气作为载体气体。这允许用户在不使用压缩气体的情况下进行麻醉。最近的研究17，18表明，使用空气代替氧气作为载体气体可能有利于许多程序。
生理监测和加热功能也可以安装到数字低流量麻醉系统中。在大多数机构中，动物变暖和生理监测需要机构动物护理和使用委员会19、20、21、22。比较麻醉剂的生理影响的研究表明，体温、心脏功能和呼吸功能23、24、25的剧烈抑郁。通常需要将动物放在加热垫上以监测和维持正常的体温。有许多动物变暖的方法，如热水器，电加热垫和热灯，但每一个都有显著的缺点。在比较不同方法的动物变暖的研究中，远红外变暖被认为是最有益的26。数字蒸发器包括内置的家热远红外变暖，以保持特定的动物体温。这消除了任何额外的加热垫控制器的需求。
除了监测体温外，脉搏氧化是监测动物心率和氧饱和度的常用方法。这种非侵入性方法简单、准确，可全面评估动物调节血液氧分水平的能力。脉冲氧化学的爪子传感器可以连接到麻醉系统，正如我们之前所证明的2。
当动物处于较长的麻醉期或需要控制动物呼吸模式时，通常需要机械通风。低流量数字蒸发器具有在压力或体积控制下提供可控呼吸的能力。集成式呼吸机无需外部呼吸机和过多的管子设置要求。
由于所有这些常见的显示器和功能都组合成一个设备，因此油管设置大大简化。此协议的目的是演示一体式数字麻醉系统的设置和使用。

<table><tbody><tr><td>Intubation Kit</td><td>Kent Scientific Corporation</td><td>ETM-MSE</td><td>Includes intubation stage, intubation tube, LED light</td></tr><tr><td>Isoflurane Liquid Inhalation 99.9%</td><td>Henry Schein, Inc.</td><td>1182097</td><td>Glass bottle 250mL</td></tr><tr><td>MouseSTAT Pulse Oximeter</td><td>Kent Scientific Corporation</td><td>SS-03</td><td>Integrated into SomnoSuite</td></tr><tr><td>Oxygen Tank</td><td>Indiana Oxygen Company</td><td>23-160246</td><td>Medical Grade O&lt;sub&gt;2&lt;/sub&gt; 99%</td></tr><tr><td>RoVent Automatic Ventilator</td><td>Kent Scientific Corporation</td><td>SS-04</td><td>Integrated into SomnoSuite</td></tr><tr><td>SomnoSuite Low Flow Digital Anesthesia System</td><td>Kent Scientific Corporation</td><td>SS-01</td><td>Includes RightTemp Homeothermic Warming control, pad, and temperature sensors</td></tr><tr><td>SomnoSuite Mouse Starter Kit</td><td>Kent Scientific Corporation</td><td>SOMNO-MSEKIT</td><td>Includes nose cone, syringes, induction chamber, and charcoal canister</td></tr></tbody></table>

use of an integrated low-flow anesthetic vaporizer, ventilator, and physiological monitoring system for rodents

低流量数字蒸发器通常使用注射器泵将挥发性麻醉剂直接插入载体气体流中。根据动物福利建议，动物在需要麻醉的程序中受到温暖和监测。常见的麻醉和生理监测设备包括煤气罐、麻醉蒸发器和支架、加热控制器和垫子、机械呼吸机和脉冲氧化计。计算机对于数据收集和运行设备软件也是必要的。在较小的空间或进行现场作业时，在有限的空间中配置所有这些设备可能具有挑战性。
该协议的目的是展示使用低流量数字蒸发器使用压缩氧气和室内空气的最佳做法，以及集成的机械呼吸机、脉冲氧化计和远红外加热，作为适合啮齿动物的全包麻醉和生理监测套件。

10周大，雄性，野生类型C57Bl6j小鼠体重25.41±0.8克用于这项研究。小鼠在鼻锥或插管上麻醉和维持，并在综合机械呼吸机上保持1.5-2.5%的异氟，同时监测心率和氧饱和度。这些动物被集体安置在微隔离箱中，并可免费获得标准啮齿动物和瓶装水。
心率和SpO2在维护过程中通过脉搏氧化（图5，图6和图7）进行监测。通过红外加热垫和热灯将体温维持在36.5-37.5°C。通风动物在插管过程中通过集成鼻锥的插管支架连续获得异氟兰的分娩。每只小鼠在鼻锥上成功通风或保持低流量，室空气（RA）或氧气（O2）不超过141毫升/分钟，持续15分钟。动物的心率和血氧饱和度保持稳定，所有组的测量几乎没有显著变化。SpO2在所有组保持在 82-99% 之间，而体温保持在 36.5-37.5 °C 之间。 我们观察到脉冲轴测仪的位置和体温都影响了SpO2的测量结果。如果我们从脉冲轴仪中观察到无效读数，我们调整了传感器的位置和加热水平，以保持核心体温稳定。
进行了双向 ANOVA 与 Bonferroni 校正，以确定 图 5、图 6 和 图 7中数据的重要性。低于0.05的 p值被认为是显著的。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/61311/61311fig01.jpg"> 图1:麻醉感应和鼻锥维护管设置图。<a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/61311/61311fig01large.jpg" target="_blank">请点击这里查看此数字的较大版本。</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/61311/61311fig02.jpg"> 图2:麻醉诱导、灌输和通风管设置图。<a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/61311/61311fig02large.jpg" target="_blank">请点击这里查看此数字的较大版本。</a>
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/61311/61311fig03.jpg"> 图3:小鼠在插管过程中通过集成鼻锥的插管支架连续获得异氟兰。<a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/61311/61311fig03large.jpg" target="_blank">请点击这里查看此数字的较大版本。</a>
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/61311/61311fig04.jpg"> 图4:集成脉冲氧化计传感器放置在后爪上。<a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/61311/61311fig04large.jpg" target="_blank">请点击这里查看此数字的较大版本。</a>
<img alt="Figure 5" class="xfigimg" src="/files/ftp_upload/61311/61311fig05.jpg"> 图5:平均心率超过15分钟±SD与房间空气（RA）或100%氧气（O2）通过鼻锥或通风通过气管（n=5/组）。没有发现不同组之间的显著差异。 <a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/61311/61311fig05large.jpg" target="_blank">请点击这里查看此数字的较大版本。</a>
<img alt="Figure 6" class="xfigimg" src="/files/ftp_upload/61311/61311fig06.jpg"> 图 6:通过低流量麻醉系统进行初始麻醉后记录的心率值 （bpm）。平均心率值在 15 分钟内从 30 秒的时间间隔计算。每个数据点表示每个组中所有动物的平均± SD （n=5）。在15分钟内，没有观察到任何组的心率有显著变化。 <a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/61311/61311fig06large.jpg" target="_blank">请点击这里查看此数字的较大版本。</a>
<img alt="Figure 7" class="xfigimg" src="/files/ftp_upload/61311/61311fig07.jpg"> 图7:组织氧饱和度水平（%）在用低流量麻醉系统进行初始麻醉后。平均 SpO2 值在 15 分钟内从 30 秒的时间间隔计算。每个数据点表示每个组中所有动物的平均± SD （n=5）。在15分钟内，在任何组中均未观察到SpO2 的显著变化。 <a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/61311/61311fig07large.jpg" target="_blank">请点击这里查看此数字的较大版本。</a>

Watch this Scientific Journal Video about Use of an Integrated Low-Flow Anesthetic Vaporizer, Ventilator, and Physiological Monitoring System for Rodents at JoVE.com

Use of an Integrated Low-Flow Anesthetic Vaporizer, Ventilator, and Physiological Monitoring System for Rodents

在这里，我们提出了一个协议，使用数字低流量麻醉系统与集成呼吸机和生理监测模块安全有效地管理麻醉气体给小鼠。

使用啮齿动物综合低流量麻醉蒸发器、呼吸机和生理监测系统

Low-flow digital vaporizers commonly utilize a syringe pump to directly administer volatile anesthetics into a stream of carrier gas. Per animal welfare ...

use-an-integrated-low-flow-anesthetic-vaporizer-ventilator

Universidad San Sebastian

Research

JoVE Journal

Medicine

5.8K 次观看.  Kent Scientific Corporation. 在这里，我们提出了一个协议，使用数字低流量麻醉系统与集成呼吸机和生理监测模块安全有效地管理麻醉气体给小鼠。

视频: Use of an Integrated Low-Flow Anesthetic Vaporizer, Ventilator, and Physiological Monitoring System for Rodents

Assessing Changes in Volatile General Anesthetic Sensitivity of Mice after Local or Systemic Pharmacological Intervention

One desirable endpoint of general anesthesia is the state of unconsciousness, also known as hypnosis. Defining the hypnotic state in animals is less straightforward than it is in human patients. A widely used behavioral surrogate for hypnosis in rodents is the loss of righting reflex (LORR), or the point at which the animal no longer responds to their innate instinct to avoid the vulnerability of dorsal recumbency. We have developed a system to assess LORR in 24 mice simultaneously while carefully controlling for potential confounds, including temperature fluctuations and varying gas flows. These chambers permit reliable assessment of anesthetic sensitivity as measured by latency to return of the righting reflex (RORR) following a fixed anesthetic exposure. Alternatively, using stepwise increases (or decreases) in anesthetic concentration, the chambers also enable determination of a population's sensitivity to induction (or emergence) as measured by EC50 and Hill slope. Finally, the controlled environmental chambers described here can be adapted for a variety of alternative uses, including inhaled delivery of other drugs, toxicology studies, and simultaneous real-time monitoring of vital signs.

Loss of the righting reflex has long served as a standard behavioral surrogate for unconsciousness, also called hypnosis, in laboratory animals. Alterations in volatile anesthetic sensitivity caused by pharmacological interventions can be detected with a carefully controlled high-throughput assessment system, which may be adapted for delivery of any inhaled therapeutic.

局部或全身药物干预后评估的变化小鼠中挥发性全身麻醉药敏感性

One desirable endpoint of general anesthesia is  the state of unconsciousness, also known as hypnosis. Defining the hypnotic state in animals is  less ...

科研

医学

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

The number of acceptable donor lungs available for lung transplantation is severely limited due to poor quality. 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. As this technology expands and improves, the ability to potentially evaluate and improve the quality of substandard lungs prior to transplant is a critical need. In order to more rigorously evaluate these approaches, a reproducible animal model needs to be established that would allow for testing of improved techniques and management of the donated lungs as well as to the lung-transplant recipient. In addition, an EVLP animal model of associated pathologies, e.g., ventilation induced lung injury (VILI), would provide a novel method to evaluate treatments for these pathologies. Here, we describe the development of a rat EVLP lung program and refinements to this method that allow for a reproducible model for future expansion. We also describe the application of this EVLP system to model VILI in rat lungs. The goal is to provide the research community with key information and &#8220;pearls of wisdom&#8221;/techniques that arose from trial and error and are critical to establishing an EVLP system that is robust and reproducible.

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.

的隔离方法<em&gt;离体</em&gt;肺灌注的大鼠模型:教训制定鼠EVLP课程中学

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

Non-restraining EEG Radiotelemetry: Epidural and Deep Intracerebral Stereotaxic EEG Electrode Placement

Implantable EEG radiotelemetry is of central relevance in the neurological characterization of transgenic mouse models of neuropsychiatric and neurodegenerative diseases as well as epilepsies. This powerful technique does not only provide valuable insights into the underlying pathophysiological mechanisms, i.e., the etiopathogenesis of CNS related diseases, it also facilitates the development of new translational, i.e., therapeutic approaches. Whereas competing techniques that make use of recorder systems used in jackets or tethered systems suffer from their unphysiological restraining to semi-restraining character, radiotelemetric EEG recordings overcome these disadvantages. Technically, implantable EEG radiotelemetry allows for precise and highly sensitive measurement of epidural and deep, intracerebral EEGs under various physiological and pathophysiological conditions. First, we present a detailed protocol of a straight forward, successful, quick and efficient technique for epidural (surface) EEG recordings resulting in high-quality electrocorticograms. Second, we demonstrate how to implant deep, intracerebral EEG electrodes, e.g., in the hippocampus (electrohippocampogram). For both approaches, a computerized 3D stereotaxic electrode implantation system is used. The radiofrequency transmitter itself is implanted into a subcutaneous pouch in both mice and rats. Special attention also has to be paid to pre-, peri- and postoperative treatment of the experimental animals. Preoperative preparation of mice and rats, suitable anesthesia as well as postoperative treatment and pain management are described in detail.

Non-restraining EEG radiotelemetry is a valuable methodological approach to record in vivo long-term electroencephalograms from freely moving rodents. This detailed protocol describes stereotaxic epidural and deep intracerebral electrode placement in different brain regions in order to obtain reliable recordings of CNS rhythmicity and CNS related behavioral stages.

非约束脑电遥测:硬膜外和脑深部脑电立体定位放置电极

Implantable EEG radiotelemetry is of central relevance in the neurological characterization of transgenic mouse models of neuropsychiatric and ...

神经科学

Use of a Low-flow Digital Anesthesia System for Mice and Rats

A traditional vaporizer depends on flowing gas and atmospheric pressure for passive anesthetic vaporization. Newly developed direct injection vaporizers utilize a syringe pump to directly administer volatile anesthetics into a gas stream. Unlike a traditional vaporizer, it can be used at very low flow rates, making it ideal for use on mice and rats. 
The equipment's capability to use low flow rates could result in a substantial cost savings due to the reduced need for anesthetic agents, compressed gas, and charcoal scavenging filters1. A lower flow rate means less waste of anesthetic gas and likely reduces the risk of anesthetic exposure to laboratory personnel. Thus, the high levels of precision and safety associated with direct injection vaporizers, along with a reduced need for anesthetic agents, compressed gas, and charcoal filters are beneficial for research requiring small animal anesthesia. 
The goal of this protocol is to demonstrate the use of a syringe-driven direct injection vaporizer as part of a digital, low-flow anesthesia system. The direct injection vaporizer is capable of accurately delivering anesthesia at very low flow rates compared to a traditional vaporizer, making it a promising alternative for controlled gas anesthetic delivery to rodents.

Here, we present a protocol to more safely and efficiently administer anesthetic gas to mice using a digital, low flow anesthesia system utilizing a syringe-driven direct injection vaporizer.

低流量数字麻醉系统的小鼠和大鼠的用途

A traditional vaporizer depends on flowing gas and atmospheric pressure for passive anesthetic vaporization. Newly developed direct injection vaporizers ...

Laminectomy and Spinal Cord Window Implantation in the Mouse

This protocol describes a method for spinal cord laminectomy and glass window implantation for in vivo imaging of the mouse spinal cord. An integrated digital vaporizer is utilized to achieve a stable plane of anesthesia at a low-flow rate of isoflurane. A single vertebral spine is removed, and a commercially available cover-glass is overlaid on a thin agarose bed. A 3D-printed plastic backplate is then affixed to the adjacent vertebral spines using tissue adhesive and dental cement. A stabilization platform is used to reduce motion artifact from respiration and heartbeat. This rapid and clamp-free method is well-suited for acute multi-photon fluorescence microscopy. Representative data are included for an application of this technique to two-photon microscopy of the spinal cord vasculature in transgenic mice expressing eGFP:Claudin-5 &#8212; a tight junction protein.

This protocol describes implantation of a glass window onto the spinal cord of a mouse to facilitate visualization by intravital microscopy.

拉米内切手术和脊髓窗口植入小鼠

This protocol describes a method for spinal cord laminectomy and glass window implantation for in vivo imaging of the mouse spinal cord. An integrated ...

Evaluation of Respiratory System Mechanics in Mice using the Forced Oscillation Technique

The forced oscillation technique (FOT) is a powerful, integrative and translational tool permitting the experimental assessment of lung function in mice in a comprehensive, detailed, precise and reproducible manner. It provides measurements of respiratory system mechanics through the analysis of pressure and volume signals acquired in reaction to predefined, small amplitude, oscillatory airflow waveforms, which are typically applied at the subject's airway opening. The present protocol details the steps required to adequately execute forced oscillation measurements in mice using a computer-controlled piston ventilator (flexiVent; SCIREQ Inc, Montreal, Qc, Canada). The description is divided into four parts: preparatory steps, mechanical ventilation, lung function measurements, and data analysis. It also includes details of how to assess airway responsiveness to inhaled methacholine in anesthetized mice, a common application of this technique which also extends to other outcomes and various lung pathologies. Measurements obtained in na&iuml;ve mice as well as from an oxidative-stress driven model of airway damage are presented to illustrate how this tool can contribute to a better characterization and understanding of studied physiological changes or disease models as well as to applications in new research areas.

The present protocol provides a detailed step-by-step description of the procedures required to execute measurements of respiratory system mechanics as well as the assessment of airway responsiveness to inhaled methacholine in mice using the forced oscillation technique (flexiVent; SCIREQ Inc, Montreal, Qc, Canada).

使用强迫振动技术在小鼠呼吸系统力学评价

The  forced oscillation technique (FOT) is a powerful, integrative and translational  tool permitting the experimental assessment of lung function in mice ...

生物学

Direct Intrabronchial Administration to Improve the Selective Agent Deposition Within the Mouse Lung

Intratracheal (IT) administration of experimental agents is an essential technique in murine models of diffuse lung diseases, such as bleomycin-induced pulmonary fibrosis.&#160; However, distribution of intratracheally-administered agents to the distal mouse lung is often asymmetric, with lung parenchymal concentrations increased in the smaller (but equally accessible) left lung of the mouse.&#160; Described in this report is a novel intrabronchial (IB) approach to cannulate the left and/or right lungs of living mice non-operatively.&#160; It is also demonstrated how this approach can be used to selectively administer agents to one lung or adapted (via dose-adjusted IB delivery) to improve the left-right symmetry of lung delivery of experimental agents, thereby improving models of diffuse lung disease such as bleomycin-induced pulmonary fibrosis.

Intratracheal (IT) administration of experimental agents in mice often results in asymmetric delivery to the distal lungs.&#160; In this report, we describe a direct intrabronchial (IB) approach to cannulate each lung in living mice non-operatively.&#160; This approach can be used to selectively administer agents to one lung or may be adapted to improve the symmetric agent delivery to both lungs.

直接注射内管理,改善小鼠肺内的选择性代理沉积

Intratracheal (IT) administration of experimental agents is an essential technique in murine models of diffuse lung diseases, such as bleomycin-induced ...

Biventricular Assessment of Cardiac Function and Pressure-Volume Loops by Closed-Chest Catheterization in Mice

Assessment of cardiac function is essential to conduct cardiovascular and pulmonary-vascular preclinical research. Pressure-volume loops (PV loops) generated by recording both pressure and volume during cardiac catheterization are vital when assessing both systolic and diastolic cardiac function. Left and right heart function are closely related, reflected in ventricular interdependence. Thus, recording biventricular function in the same animal is important to get a complete assessment of cardiac function. In this protocol, a closed chest approach to cardiac catheterization consistent with the way catheterization is performed in patients is adopted in mice. While challenging, the closed chest strategy is a more physiological approach, because opening the chest results in major changes in preload and afterload that create artifacts, most notably a fall in systemic blood pressure. While high-resolution echocardiography is used to assess rodents, cardiac catheterization is invaluable, particularly when assessing diastolic pressures in both ventricles.
Described here is a procedure to perform invasive, closed chest, sequential left and right ventricular pressure-volume (PV) loops in the same animal. PV loops are acquired using&#160;admittance technology with a mouse pressure-volume catheter and pressure-volume system acquisition. The procedure is described, beginning with the neck dissection, which is required to access the right jugular vein and the right carotid artery, to the insertion and positioning of the catheter, and finally the data acquisition. Then, the criteria required to ensure the acquisition of high-quality PV loops are discussed. Finally, the analysis of the left and right ventricular PV loops and the different hemodynamic parameters available to quantify systolic and diastolic ventricular function are briefly described.

Presented here is a protocol to assess biventricular heart function in mice by generating pressure-volume (PV) loops from the right and left ventricle in the same animal using closed chest catheterization. The focus is on the technical aspect of surgery and data acquisition.

小鼠闭胸导管插入术对心功能和压力-容积环的双心室评估

Assessment of cardiac function is essential to conduct cardiovascular and pulmonary-vascular preclinical research. Pressure-volume loops (PV loops) ...

Measuring Pressure Volume Loops in the Mouse

Understanding the causes and progression of heart disease presents a significant challenge to the biomedical community. The genetic flexibility of the mouse provides great potential to explore cardiac function at the molecular level. The mouse&#39;s small size does present some challenges in regards to performing detailed cardiac phenotyping. Miniaturization and other advancements in technology have made many methods of cardiac assessment possible in the mouse. Of these, the simultaneous collection of pressure and volume data provides a detailed picture of cardiac function that is not available through any other modality. Here a detailed procedure for the collection of pressure-volume loop data is described. Included is a discussion of the principles underlying the measurements and the potential sources of error. Anesthetic management and surgical approaches are discussed in great detail as they are both critical to obtaining high quality hemodynamic measurements. The principles of hemodynamic protocol development and relevant aspects of data analysis are also addressed.

This manuscript describes a detailed protocol for the collection of pressure-volume data from the mouse.

测量压力容量环中的鼠标

Understanding the causes and progression of heart disease presents a significant challenge to the biomedical community. The genetic flexibility of the ...

使用倒置显微镜进行活体成像以跟踪皮肤细胞动力学

Streamlined Intravital Imaging Approach for Long-Term Monitoring of Epithelial Tissue Dynamics on an Inverted Confocal Microscope

Understanding normal and aberrant in vivo cell behaviors is necessary to develop clinical interventions to thwart disease initiation and progression. It is therefore critical to optimize imaging approaches that facilitate the observation of cell dynamics in situ, where tissue structure and composition remain unperturbed. The epidermis is the body&#39;s outermost barrier, as well as the source of the most prevalent human cancers, namely cutaneous skin carcinomas. The accessibility of skin tissue presents a unique opportunity to monitor epithelial and dermal cell behaviors in intact animals using noninvasive intravital microscopy. Nevertheless, this sophisticated imaging approach has primarily been achieved using upright multiphoton microscopes, which represent a significant barrier for entry for most investigators. This study presents a custom-designed, 3D-printed microscope stage insert suitable for use with inverted confocal microscopes, streamlining the long-term intravital imaging of ear skin in live transgenic mice. We believe this versatile invention, which may be customized to fit the inverted microscope brand and model of choice and adapted to image additional organ systems, will prove invaluable to the greater scientific research community by significantly enhancing the accessibility of intravital microscopy. This technological advancement is critical for bolstering our understanding of live cell dynamics in normal and disease contexts.

作者聚焦：开发使用倒置共聚焦显微镜进行体内活体成像的工具 

The protocol presents a new tool to simplify intravital imaging using inverted confocal microscopy.

简化活体成像方法，用于在倒置共聚焦显微镜上长期监测上皮组织动力学

Understanding normal and aberrant in vivo cell behaviors is necessary to develop clinical interventions to thwart disease initiation and progression. It ...