Name: use of a low-flow digital anesthesia system for mice and rats
Uploaded: 2016-09-07T13:00:00.000+00:00
Duration: 8 min 6 s
Description: Watch this Scientific Journal Video about Use of a Low-flow Digital Anesthesia System for Mice and Rats at JoVE.com

The authors have no acknowledgements.

所有研究均按照监管和制度指引完成。这项研究的动物方面是由肯特科技公司使用动物计划，由普渡大学动物护理和使用委员会（PACUC）认可的评估，并按照指南实验动物22的管理和使用进行。 注:本协议所使用的低流量数字麻醉系统配备有集成脉搏血氧饱和度。 1.建立集成式脉搏血氧仪低流量麻醉系统<ol><li>异氟醚交货<ol><li>选择载气源。要利用内部空气泵，旋进气口上背部，使内部泵吸入室内空气。 </li><li>连接活性炭罐到排气口。 </li><li> Y型适配器连接到低流量，数字麻醉系统的前面。使用颜色编码剪辑连接白色分支到t他鼻锥和蓝色分支到感应腔室。 </li><li>关闭白色夹子夹并打开蓝色夹子夹直接的气流室。 </li><li>选择2毫升注射器。 </li></ol></li><li>对于生理监测，使用集成式脉搏血氧仪<ol><li>连接脉搏血氧计爪子传感器端口上的低流量麻醉系统的背面，标有"AUX"。 </li></ol></li></ol> 2.配置设置<ol><li>麻醉<ol><li>电力麻醉系统，并进入设置菜单。按设置访问红色主菜单 &gt; 麻醉 &gt; 设置 。 </li><li>选择麻醉剂。按设置为突出型ANEST为红色。使用向上和向下箭头选择异氟醚。 </li><li>设置注射器大小。按设置以突出锡尔河大小 。使用向上和向下箭头选择2毫升注射器。 </li><li>设置空位。按设置为highlight红色设置空 。通过将注射器使注射器保持夹具上的注射器的金属套环就位固定在注射器保持块为空，完全压下玻璃注射器。按向上或向下箭头移动推块使推块使得与注射器活塞顶部的轻微接触。按选择设置空位。 </li><li>按设置以突出红色的删除 。取下注射器和填充用的瓶顶适配器异氟醚注射器。 </li><li>注射器连接到麻醉系统。 </li><li>总理麻醉递送管道。按设置以突出红色坯管 。向下按，直到麻醉穿过注射器进入蒸发器模块上的黑色配件。 </li><li>启用麻醉。按SET UP键突出显示启用红色。使用向上和向下箭头选择是 。按Run /返回，返回到主菜单。 </li><li>选择一个IR供应和每分钟通气量。按SET UP突出供气红色。使用向上和向下箭头键选择内置泵 。按设置以突出红色的分卷 。设定流速250毫升/分钟。 </li></ol></li><li>生理监控<ol><li>设定的最低检测心脏速率鼠标（240）。按SET UP键进入主菜单 &gt; MouseStat，用上下箭头设定最低心脏速率。 </li></ol></li></ol> 3.开始麻醉系统<ol><li>诱导鼠标<ol><li>按Run /返回两次进入运行模式，并开始气流。 </li><li>放置在感应室鼠标，关闭盖子紧。调节麻醉剂浓度旋钮3％。 </li><li>监测直到鼠标已经达到麻醉的所需平面，通过翻正反射损失决定。调节麻醉剂浓度是必要的。 </li><li>一旦动物失去其翻正反射一ND充分麻醉，打开麻醉浓度表盘为0％。笔者以前曾发现，让气流冲感应室30-60秒就足以清除室没有扭转麻醉深度1。 </li><li>快速打开白色夹子的空气直接到面罩，并关闭蓝色夹子导致感应室。 </li><li>从研究者打开密室走，移开鼠标，并立即适应鼻锥。 </li><li>围绕红外变暖垫，设置经由反馈回路直肠探头以维持体温在37℃的动物。 </li><li>当鼠标在鼻锥稳定，调整异氟烷的浓度至1.5％，或根据需要进行维护通过转动麻醉剂浓度旋钮。 </li><li>减少分钟通气量进行​​维护。的最小流量，以支持动物等于动物的分钟量1.5-2.2倍（在30 G鼠标，最小52毫升/分钟）。指制造商的说明为一个推荐流量具体设定为鼻锥样式，并根据需要进行调整。按SET UP键进入主菜单，然后按设定键直到分卷以红色突出显示。使用向上和向下箭头调整目标流量。按Run /返回，返回到主屏幕。 </li><li>确认麻醉深度为叉指捏过程中缺乏撤退反射的决定。应用眼药膏的眼睛，以防止在麻醉过程中干燥。 </li></ol></li></ol> 4.开始机能监测<ol><li>将传感器放到后掌垫。从而使红灯爪子底下，照亮了爪子传感器的位置。使用屏幕上的向上和向下箭头以显示oxiwave。动物现在已经安全使用低流量，注射器驱动，数字蒸发器麻醉。 </li></ol> 5.取下动物<ol><li>关闭麻醉系统。 <ol><li>要停止麻醉剂的递​​送，把麻醉剂浓度旋钮民（或0％）和口罩取下动物。 </li><li>监控麻醉恢复过程中的鼠标。一旦鼠标已经完全卧床，使其返回到笼子里。 </li></ol></li></ol>

<ol>
	<li>Damen, F. W., Adelsperger, A. R., Wilson, K. E., Goergen, C. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+traditional+and+integrated+digital+anesthetic+vaporizers.">Comparison of traditional and integrated digital anesthetic vaporizers.</a> JAALAS. 54 (6), 756-762 (2015).</li><li>Chakravarti, S., Basu, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modern+Anaesthesia+Vapourisers.">Modern Anaesthesia Vapourisers.</a> Indian J Anaesth. 57 (5), 464-471 (2013).</li><li>Carroll, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Small Animal Anesthesia and Analgesia. , (2008).</li><li>Tranquilli, W. J., Thurmon, J. C., Grimm, K. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Lumb and Jones' veterinary anesthesia and analgesia. , 23-86 (2013).</li><li>Ambrisko, T. D., Klide, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+isoflurane+and+Sevoflurane+vaporizers+over+a+wide+range+of+oxygen+flow+rates.">Evaluation of isoflurane and Sevoflurane vaporizers over a wide range of oxygen flow rates.</a> Am J Vet Res. 67 (6), 936-940 (2006).</li><li>McKelvey, D. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Veterinary Anesthesia and Analgesia. , (2003).</li><li>Thomas, J., Lerche, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Anesthesia and Analgesia for Veterinary Technicians. , 335 (2011).</li><li>Flecknell, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Laboratory animal anaesthesia. , (2009).</li><li>Mapleson, W. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+elimination+of+rebreathing+in+various+semiclosed+anaesthetic+systems.">The elimination of rebreathing in various semiclosed anaesthetic systems.</a> Brit J Anaesth. 26 (5), 323-332 (1954).</li><li>Ward, C. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Physical+principles+and+maintenance.">Physical principles and maintenance.</a> Anaesthetic equipment. , (1985).</li><li>El-Attar, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Guided+isoflurane+injection+in+a+totally+closed+circuit.">Guided isoflurane injection in a totally closed circuit.</a> Anaesthesia. 46 (12), 1059-1063 (1991).</li><li>Lockwood, G., Chakrabarti, M. K., Whitwam, 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=A+computer-controller+closed+anaesthetic+breathing+system.">A computer-controller closed anaesthetic breathing system.</a> Anaesthesia. 48 (8), 690-693 (1993).</li><li>Lowe, H. J., Cupic, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dose-regulated+automated+anesthesia+(Abstract).">Dose-regulated automated anesthesia (Abstract).</a> Br. J. Clin. Pharmacol. 12 (2), 281-282 (1971).</li><li>Soro, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+accuracy+of+the+anesthetic+conserving+device+(Anaconda)+as+an+alternative+to+the+classical+vaporizer+in+anesthesia.">The accuracy of the anesthetic conserving device (Anaconda) as an alternative to the classical vaporizer in anesthesia.</a> Anes Analg. 111 (5), 1176-1179 (2010).</li><li>Walker, T. J., Chackrabarti, M. K., Lockwood, G. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Uptake+of+desflurane+during+anaesthesia.">Uptake of desflurane during anaesthesia.</a> Anaesthesia. 51 (1), 33-36 (1996).</li><li>Weingarten, M., Lowe, H. 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+new+circuit+injection+technic+for+syringe-measured+administration+of+methoxyflurane:+a+new+dimension+in+anesthesia.">A new circuit injection technic for syringe-measured administration of methoxyflurane: a new dimension in anesthesia.</a> Anes Analg. 52 (4), 634-642 (1973).</li><li>Enlund, M., Wiklund, L., Lambert, H. <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+device+to+reduce+the+consumption+of+a+halogenated+anaesthetic+agent.">A new device to reduce the consumption of a halogenated anaesthetic agent.</a> Anaesthesia. 56 (5), 429-432 (2001).</li><li>Kelly, J. M., Kong, K. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accuracy+of+ten+isoflurane+vaporisers+in+current+clinical+use.">Accuracy of ten isoflurane vaporisers in current clinical use.</a> Anaesthesia. 66 (8), 682-688 (2011).</li><li>Matsuda, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=NARCOBIT+-+A+newly+developed+inhalational+anesthesia+system+for+mice.">NARCOBIT - A newly developed inhalational anesthesia system for mice.</a> Exp Anim. 56 (2), 131-137 (2007).</li><li>Matsuda, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+newly+developed+inhalation+anesthesia+system+and+intraperitoneal+anesthesia+on+the+hemodynamic+state+in+mice.">Comparison of newly developed inhalation anesthesia system and intraperitoneal anesthesia on the hemodynamic state in mice.</a> Biol Pharm Bull. 30 (9), 1716-1720 (2007).</li><li>Voightsverger, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sevoflurane+ameliorates+gas+exchange+and+attenuates+lung+damage+in+experimental+lipopolysaccharide-induced+lung+injury.">Sevoflurane ameliorates gas exchange and attenuates lung damage in experimental lipopolysaccharide-induced lung injury.</a> Anesthesiology. 111 (6), 1238-1248 (2009).</li><li>Garber, 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=.">.</a> Guide for the Care and Use of Laboratory Animals. , (2011).</li></ol>

该项目由肯特科技公司，美国心脏协会CJ Goergen指出（SDG18220010）和普渡大学提供设备和资金的支持。作者克丽斯塔Bigiarelli和伊琳娜Toore是产生在这篇文章中使用的设备肯特科技公司的员工。这篇文章的开放获取出版由肯特科技公司赞助。

数字低流量麻醉系统允许用户有效地麻醉在非常低流速的小鼠，而无需使用任何压缩气体。这从标准无源汽化器，其中大部分需要在约500毫升/分钟的最小流速一个压缩气体源有很大不同。标准蒸发器利用缺乏层次之间的精确刻度盘，他们必须每年维修，以保持精度。注射器驱动麻醉剂系统可以在设定流量提供麻醉剂的特定浓度来计算所述注射器泵的确切需要的速度。常规标定是不必要的，从而导致额外的成本和节省时间。 建议的最低流量，以维持在非再呼吸电路的动物是1.5-2.2倍的动物的分钟量。在这项研究中使用了100毫升/分钟的流速超过了这个最小，以传递足够的麻醉的动物。流速设置搜索GS是这种麻醉剂递送技术关键的，作为流量直接相关的异氟烷用于给定的时间内的量。当在低流速时，这种技术可以大大降低使用过程中所需的异氟醚的量，而动物仍然有效1,19-21麻醉。 传统的蒸发器和低流量数字蒸发器之间的新设备的成本是相当的。然而，数字低流量麻醉系统具有提供任一异氟烷或七氟醚的能力。这消除了对指定异氟醚和七氟醚精度汽化器，减少了同时使用麻醉剂组初始设备成本的需求。蒸发器技术之间最近发表的比较都使用低流量数字蒸发器1,19,20时提出节约成本随着时间的推移。这些比较的结果可用于在一年的过程中潜在的成本节约近似。如舒米恩在2小时的增量，每周5天的52周进行典型使用设置，传统的异氟烷蒸发器将消耗3.8升异氟烷，或12个250 ml瓶装。在相同的频率中使用的低流量数字汽化器将消耗只是0.32 L或二份各250ml的瓶子。活性碳罐的消耗也减少了。假设每个罐持有50克清理废气，传统的蒸发器，将填补约21木炭罐超过一年的课程。相比之下，低流量数字汽化器将需要6个或更少。传统的蒸发器将需要每年约5大气瓶，每9,500 L的内置气泵，在数字低流量汽化器某些型号的容量，省去了压缩气体的要求。如果压缩气体都被使用，则系统将使用每年1只1气缸。 该技术可以根据需要进行修改。低流量数字vaporiz器允许用户快速和精确地调整麻醉深度。如果麻醉深度必须增加或减小时，用户​​可以使用拨号的系统的顶部增加0.1％的增量麻醉浓度。根据需要在整个过程也可调节的流速。该协议采用2毫升注射器，但更大的注射器规格可供较长的过程。内部空气泵为用户提供了麻醉动物，而不需要一个压缩气体源的选项。对于需要压缩气体或补充氧气程序时，用户具有对气体源到低流量系统连接，而不是使用周围空气的选项。用户可以继续对整个过程提供所选择的空气源，或可以在内部泵并根据需要在压缩气体源之间切换。例如，用户可将系统设置为诱导和维持期间通过内部泵输送室内空气，但提供补充氧恢复期间根。 虽然有很多优点，使用低流量数字蒸发器，也有局限性。因为冲洗阀不包括，开口之前手动冲洗用清洁空气腔室是吹扫感应腔室的唯一途径。这个系统被设计成在仅低流速操作和不提供上述800毫升/分钟，而传统的蒸发器可与流率被用于高达10升/分钟流速麻醉。因此，这种特定的系统是仅适用于小动物物种。另外，相对于传统的蒸发器系统中保持较少的麻醉剂。可能存在的注射器必须在手术期间再填充的情况。然而，可以通过填写预第二注射​​器附近，以取代空注射器可​​以减少再填充期间的延迟。注射器尺寸可达10 mL可用来减少需要重新填充注射器中端程序。最后，与传统的蒸发器中，低FL流数字蒸发器需要的电力。电池是可用于实例使用，其中电力不可用或在停电的情况下。 以前的研究已经表明，低流量数字系统相比，传统的麻醉系统1,19,20消耗更少的异氟烷，载气，和木炭罐。清除的麻醉气体的减少也能识别垃圾麻醉气体的减少，虽然需要在这些领域进一步开展工作。红外气体光谱可用于监测废物异氟醚的生产，和剂量计徽章可以用于量化在将来的比较异氟烷暴露于实验室人员。 总之，该技术对于麻醉剂递送将是执行啮齿动物麻醉由于改善的安全性，有效性和精度比传统系统组是有益的。

有迹象表明，0.5-10升/分钟2流量之间的操作可兽用许多精密喷雾器。这些流量是不理想的老鼠相比，它们的小每分钟呼吸量范围为高。高流速不会在兽医实践建议由于它们促进低温和呼吸道3,4-干燥的。此外，许多普通兽医汽化器制造商的手册警告高流速可能导致背压的波动的发生增加。还已经表明，许多标准汽化器变得不准确低于500ml /分的流速，和该速率被认为是在兽医领域5-7最小流率。 动物可以保持在T形件的电路，或使用低至1.5-2.2倍动物的分钟量8-10流速改性贝恩电路。这些流量被认为是sufficien吨至到期防止气体再呼吸，防止血液中二氧化碳浓度8的增加。使用该流速建议，30 G鼠标可维持在流速低高达52毫升/分钟，高于最低传统蒸发器的接受500ml /分少了近十倍。 而传统的汽化器取决于气流并用于被动麻醉剂汽化大气压，直喷汽化器测量总新鲜气体流并直接注入蒸汽到气体流2。一些直接喷射喷雾器利用注射器泵施用麻醉剂到气体流中。计算机化控制允许这些系统来自动调节注射泵的速度来注入达到麻醉剂的所需浓度所需的液体试剂的量。注射器驱动的蒸发器可用，并批准用于临床和儿童使用，许多类似的配置视为美学在临床实践中11-16节约型的设备。他们的批准后不久，随着注射泵喷雾器麻醉节约设备被改编为在动物研究8,17,18使用。不同于传统的蒸发器，利用一个注射泵直接喷射系统不是由最小流率，以保持精度的限制。出于这个原因，这种技术非常适用于啮齿动物麻醉并在低流率是必要的其它实例中使用。与此蒸发器设计相关的益处和潜在的成本节约启发专门为啮齿类1,19,20设计新麻醉系统的发展。这种新的系统还包括一个内置的气泵，允许用户管理麻醉而不需要压缩气体源。作为一个额外的好处，该系统是预先校准为与两个异氟醚和七氟醚的使用。凭借在实验动物领域引进这项技术的蒸发器，我t是现在有可能在流速接近推荐的水平麻醉小实验室动物，而不需要压缩气体。

<table><tbody><tr><td>&lt;em&gt;Anesthetic Equipment&lt;/em&gt;</td><td></td><td></td><td></td></tr><tr><td>SomnoSuite Low-Flow Digital Anesthesia System</td><td>Kent Scientific Corporation</td><td>SOMNO</td><td>Includes anti-spill, anti-vapor bottle top adapter; Y adapter tubing; charcoal scavenging filter</td></tr><tr><td>MouseSTAT Pulse Oximeter &amp;amp; Heart Rate Monitor</td><td>Kent Scientific Corporation</td><td>SS-MSTAT-Module</td><td>Integrated into SomnoSuite</td></tr><tr><td>MouseSTAT Mouse Paw Sensor</td><td>Kent Scientific Corporation</td><td>MSTAT-MSE</td><td></td></tr><tr><td>2 ml Glass Syringe</td><td>Kent Scientific Corporation</td><td>SOMNO-2ML</td><td></td></tr><tr><td>Low-Cost Induction Chamber, 0.5 L</td><td>Kent Scientific Corporation</td><td>SOMNO-0705</td><td></td></tr><tr><td>Low Profile Facemask, x-small</td><td>Kent Scientific Corporation</td><td>SOMNO-0304</td><td></td></tr><tr><td>&lt;em&gt;Animal Warming&lt;/em&gt;</td><td></td><td></td><td></td></tr><tr><td>PhysioSuite Physiological Monitoring System with RightTemp Homeothermic Warming</td><td>Kent Scientific Corporation</td><td>PS-RT</td><td>Includes infrared warming pad, rectal probe, and pad temperature probe</td></tr><tr><td>&lt;em&gt;Anesthetic Agents&lt;br/&gt; and Medications&lt;/em&gt;</td><td></td><td></td><td></td></tr><tr><td>Isoflurane (250 ml bottle)</td><td>Piramal Healthcare</td><td></td><td></td></tr><tr><td>Puralube Opthalmic Ointment</td><td>Perrigo</td><td></td><td></td></tr></tbody></table>

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.

动物 3成年C57 / BL6NTac雌性小鼠（Taconic，年龄6-7周; 15重量+/- 1克）麻醉，并以1.3-1.5％异氟烷维持而心脏速率，氧饱和度，和呼吸速率进行了监测。所有小鼠的鼠无病原体的常规供应商测试抵达设施之前所确定。这些动物群养在了显微笼养并提供了一​​瓶免费获取标准啮齿动物食物和水。 使用异氟醚低流量麻醉系统测量在使用期间残留在注射器麻醉剂的量。注射器中的体积，如通过麻醉系统测量，指出作为动物在维持期结束时转移到鼻锥，并再次。最终体积W从初始体积减去量化维修期间（ 图1）所消耗的麻醉剂的量。 生理参数心脏率， 血氧饱和度和呼吸速率通过脉搏血氧饱和度（ - 4 图2）维护期间进行了监测。体温是通过一个红外暖垫保持在37.5℃。每只小鼠成功地保持以100ml室内空气的/分低流速麻醉的手术平面下60分钟后，通过从一个交叉指捏缺乏撤退反射的测定。鼠标没有清醒或在维修期间间歇应用趾间捏回应。动物的心脏速率（ 图2），血氧（ 图3），和呼吸率（ 图4）保持RELAT整个研究ively稳定。由于动物和传感器的定位，从鼠标1和鼠标3的呼吸速率的信号是间歇性的，并测量被中断。当动物的定位调整，信号改善，所测量的呼吸率是可比的他人在类似的时间点。低流量数字麻醉系统维护期间（ 图1）的60分钟期间使用的0.63毫升异氟醚的平均值。 <img alt="图1" src="/files/ftp_upload/54436/54436fig1.jpg" /> 图 1: 异氟醚异氟醚的使用在毫升超过使用数字低流量麻醉系统麻醉维持1小时用于三种不同的小鼠量<a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/54436/54436fig1large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> 图 2: 心率三只老鼠在每分钟（BPM）的初始麻醉诱导与数字低流量麻醉系统后5-60分钟节拍心脏速率<a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/54436/54436fig2large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图3" src="/files/ftp_upload/54436/54436fig3.jpg" /> 图3:。 血氧饱和度三只老鼠初始麻醉诱导与数字低流量麻醉系统后5-60分钟的血氧饱和度（％） <a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/54436/54436fig3large.jpg" target="_blank">点击此处查看该图的放大版本。</a> 图4:。呼吸率三只老鼠在每分钟（BPM）的初始麻醉诱导与数字低流量麻醉系统后5-60分钟呼吸的呼吸速率<a href="https://www-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/54436/54436fig4large.jpg" target="_blank">请点击此处查看该图的放大版本。</a>

Watch this Scientific Journal Video about Use of a Low-flow Digital Anesthesia System for Mice and Rats at JoVE.com

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

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 ...

use-of-a-low-flow-digital-anesthesia-system-for-mice-and-rats

Universidad San Sebastian

Research

JoVE Journal

Medicine

17.8K 次观看.  Purdue University. 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. 

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

Direct Intraventricular Delivery of Drugs to the Rodent Central Nervous System

Due to an inability to cross the blood brain barrier, certain drugs need to be directly delivered into the central nervous system (CNS). Our lab focuses specifically on antisense oligonucleotides (ASOs), though the techniques shown in the video here can also be used to deliver a plethora of other drugs to the CNS. Antisense oligonucleotides (ASOs) have the capability to knockdown sequence-specific targets 1 as well as shift isoform ratios of specific genes 2. To achieve widespread gene knockdown or splicing in the CNS of mice, the ASOs can be delivered into the brain using two separate routes of administration, both of which we demonstrate in the video.
The first uses Alzet osmotic pumps, connected to a catheter that is surgically implanted into the lateral ventricle. This allows the ASOs to be continuously infused into the CNS for a designated period of time. The second involves a single bolus injection of a high concentration of ASO into the right lateral ventricle. Both methods use the mouse cerebral ventricular system to deliver the ASO to the entire brain and spinal cord, though depending on the needs of the study, one method may be preferred over the other.

We describe a method to target drugs to the central nervous system by either implanting a catheter or performing a bolus injection into the right lateral ventricle in mice. We focus specifically on the delivery of antisense oligonucleotides. This technique is readily adaptable to other drugs and to rats.

啮齿动物的中枢神经系统药物直接脑室交货

Due to an inability to cross the blood brain barrier, certain drugs need to be directly delivered into the central nervous system (CNS). Our lab focuses ...

科研

神经科学

Proper Positioning and Restraint of a Rat Hind Limb for Focused High Resolution Imaging of Bone Micro-architecture Using In Vivo Micro-computed Tomography

The use of in vivo micro-computed tomography (&#181;CT) is a powerful tool which involves the non-destructive imaging of internal structures at high resolutions in live animal models. This allows for repeated imaging of the same rodent over time. This feature not only reduces the total number of rodents required in an experimental design and thereby reduces the inter-subject variation that can arise, but also allows researchers to assess longitudinal or life-long responses to an intervention. To acquire high quality images that can be processed and analyzed to more accurately quantify outcomes of bone micro-architecture, users of in vivo &#181;CT scanners must properly anesthetize the rat, and position and restrain the hind limb. To do this, it is imperative that the rat be anesthetized to a level of complete relaxation, and that pedal reflexes are lost. These guidelines may be modified for each individual rat, as the rate of isoflurane metabolism can vary depending on strain and body size. Proper technique for in vivo &#181;CT image acquisition enables accurate and consistent measurement of bone micro-architecture within and across studies.

Proper Positioning and Restraint of a Rat Hind Limb for Focused High Resolution Imaging of Bone Micro-architecture Using In Vivo Micro-computed Tomography

This paper instructs users of in vivo micro-computed tomography (&#181;CT) scanners how to anesthetize, correctly position and restrain the hind limb of a rat for minimal movement during high-resolution imaging of the tibia. The result is high quality images that can be processed to accurately quantify bone micro-architecture.

应用活体显微计算机断层成像技术对骨微结构高分辨成像大鼠后肢的定位与抑制

The use of in vivo micro-computed tomography (&#181;CT) is a powerful tool which involves the non-destructive imaging of internal structures at high ...

生物工程

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 ...

Cavernous Nerve Stimulation and Recording of Intracavernous Pressure in a Rat

The stimulation of the cavernous nerve (CN) and measurement of intracavernous pressure (ICP) have been used extensively to test and evaluate therapies for erectile dysfunction. However, the methods used vary between laboratories, and pitfalls still exist. The goal of this study was to describe a surgical technique that would provide a reliable and reproducible model. By exposing the ischiocavernosus muscle at its point of insertion on the ischial tuberosity, the penile crus could be cannulated with minimal dissection and injury to the structures involved in erectile function. Repeated stimulation of the CN, without the need for lifting and drying, was achieved by using a 125 &#181;m bipolar silver electrode and biocompatible silicon glue to isolate the electrode-nerve complex. This method prevents neuropraxia by reducing stretching and drying the nerve and provides complete isolation of the nerve, negating electrical leakage and preventing stimulation of alternative pathways.

This study describes a simplified surgical procedure and technique for performing cavernous nerve stimulation with the isolation of the nerve-electrode complex using silicone glue and intracavernous pressure measurement.

大鼠海绵状神经刺激与海绵压力记录

The stimulation of the cavernous nerve (CN) and measurement of intracavernous pressure (ICP) have been used extensively to test and evaluate therapies for ...

医学

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 ...

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 ...

In vivo Calcium Imaging in Mouse Inferior Olive

Inferior olive (IO), a nucleus in the ventral medulla, is the only source of climbing fibers that form one of the two input pathways entering the cerebellum. IO has long been proposed to be crucial for motor control and its activity is currently considered to be at the center of many hypotheses of both motor and cognitive functions of the cerebellum. While its physiology and function have been relatively well studied on single-cell level in vitro, presently there are no reports on the organization of the IO network activity in living animals. This is largely due to the extremely challenging anatomical location of the IO, making it difficult to subject to conventional fluorescent imaging methods, where an optic path must be created through the entire brain located dorsally to the region of interest.
Here we describe an alternative method for obtaining state-of-the-art -level calcium imaging data from the IO network. The method takes advantage of the extreme ventral location of the IO and involves a surgical procedure for inserting a gradient-refractive index (GRIN) lens through the neck viscera to come into contact with the ventral surface of the calcium sensor GCaMP6s-expressing IO in anesthetized mice. A representative calcium imaging recording is shown to demonstrate the feasibility to record IO neuron activity after the surgery. While this is a non-survival surgery and the recordings must be conducted under anesthesia, it avoids damage to life-critical brainstem nuclei and allows conducting large variety of experiments investigating spatiotemporal activity patterns and input integration in the IO. This procedure with modifications could be used for recordings in other, adjacent regions of the ventral brainstem.

In vivo Calcium Imaging in Mouse Inferior Olive

We present a protocol to expose the brainstem of adult mouse from the ventral side. By using a gradient-refractive index lens with a miniature microscope, calcium imaging can be used to examine the activity of inferior olive neural somata in vivo.

在体内 小鼠劣质橄榄中的钙成像

Inferior olive (IO), a nucleus in the ventral medulla, is the only source of climbing fibers that form one of the two input pathways entering the ...

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

Low-flow digital vaporizers commonly utilize a syringe pump to directly administer volatile anesthetics into a stream of carrier gas. Per animal welfare recommendations, animals are warmed and monitored during procedures requiring anesthesia. Common anesthesia and physiological monitoring equipment include gas tanks, anesthetic vaporizers and stands, warming controllers and pads, mechanical ventilators, and pulse oximeters. A computer is also necessary for data collection and to run equipment software. In smaller spaces or when performing field work, it can be challenging to configure all this equipment in limited space.
The goal of this protocol is to demonstrate best practices for use of a low-flow digital vaporizer using both compressed oxygen and room air, along with an integrated mechanical ventilator, pulse oximeter, and far infrared warming as an all-inclusive anesthesia and physiological monitoring suite ideal for rodents.

Here, we present a protocol to safely and effectively administer anesthetic gas to mice using a digital, low flow anesthesia system with integrated ventilator and physiological monitoring modules.

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 ...

Intracerebroventricular Delivery of Gut-Derived Microbial Metabolites in Freely Moving Mice

The impact of gut microbiota and their metabolites on host physiology and behavior has been extensively investigated in this decade. Numerous studies have revealed that gut microbiota-derived metabolites modulate brain-mediated physiological functions through intricate gut-brain pathways in the host. Short-chain fatty acids (SCFAs) are the major bacteria-derived metabolites produced during dietary fiber fermentation by the gut microbiome. Secreted SCFAs from the gut can act at multiple sites in the periphery, affecting the immune, endocrine, and neural responses due to the vast distribution of SCFAs receptors. Therefore, it is challenging to differentiate the central and peripheral effects of SCFAs through oral and intraperitoneal administration of SCFAs. This paper presents a video-based method to interrogate the functional role of SCFAs in the brain via a guide cannula in freely moving mice. The amount and type of SCFAs in the brain can be adjusted by controlling the infusion volume and rate. This method can provide scientists with a way to appreciate the role of gut-derived metabolites in the brain.

Gut-derived microbial metabolites have multifaceted effects leading to complex behavior in animals. We aim to provide a step-by-step method to delineate the effects of gut-derived microbial metabolites in the brain via intracerebroventricular delivery via a guide cannula.

自由移动小鼠中肠道来源的微生物代谢物的脑室内递送

The impact of gut microbiota and their metabolites on host physiology and behavior has been extensively investigated in this decade. Numerous studies have ...

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 ...

低流量数字麻醉系统的小鼠和大鼠的用途

摘要

探索更多视频

此视频中的章节

啮齿动物的中枢神经系统药物直接脑室交货

应用活体显微计算机断层成像技术对骨微结构高分辨成像大鼠后肢的定位与抑制

非约束脑电遥测:硬膜外和脑深部脑电立体定位放置电极

大鼠海绵状神经刺激与海绵压力记录

拉米内切手术和脊髓窗口植入小鼠

直接注射内管理,改善小鼠肺内的选择性代理沉积

在体内小鼠劣质橄榄中的钙成像

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

自由移动小鼠中肠道来源的微生物代谢物的脑室内递送

测量压力容量环中的鼠标

低流量数字麻醉系统的小鼠和大鼠的用途

摘要

探索更多视频

此视频中的章节

啮齿动物的中枢神经系统药物直接脑室交货

应用活体显微计算机断层成像技术对骨微结构高分辨成像大鼠后肢的定位与抑制

非约束脑电遥测:硬膜外和脑深部脑电立体定位放置电极

大鼠海绵状神经刺激与海绵压力记录

拉米内切手术和脊髓窗口植入小鼠

直接注射内管理,改善小鼠肺内的选择性代理沉积

在体内 小鼠劣质橄榄中的钙成像

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

自由移动小鼠中肠道来源的微生物代谢物的脑室内递送

测量压力容量环中的鼠标

在体内小鼠劣质橄榄中的钙成像