Name: fabrication and implantation of miniature dual-element strain gages for measuring in vivo gastrointestinal contractions in rodents.
Uploaded: 2014-09-18T14:00:00.000+00:00
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Description: Watch this Scientific Journal Video about Fabrication and Implantation of Miniature Dual-element Strain Gages for Measuring In Vivo Gastrointestinal Contractions in Rodents. at JoVE.com

科研经费是通过研究所收到的神经疾病和中风（NS049177和NS087834）的。作者要感谢已故的保罗博士巴斯和他的同事对应变计的原设计的知识贡献;和卡罗尔Tollefsrud为应变片的制造和销售，直到生产在2010年和她的见地通信停止。

所有程序遵循健康指引全国学院和医学院的宾夕法尼亚州立大学贺喜批准的机构动物护理和使用委员会。大鼠用普通动物饲养规范安置。注意:该协议采用雄性Wistar大鼠≥8周龄和初始体重175 - 200克 1，程序应变仪的研制<ol><li>大多数工具和部件仍可以从原始或后继公司和总结于表1中 。 </li><li>准备和两个单元素应变计粘接<ol><li>处理应变元素（EA-06-031-350）小心地用干净的杜蒙＃5镊子。限制元件的不希望的运动，可以使用一个小的，清洁的，自粘一张纸与粘接面朝上，以确保元件的工作表面，而没有污染或过度密合性的危险。 </li><li>债券两个单应变计内容背到后面，形成一个双元件。清洗各元件薄膜的背面与异丙醇，并允许干燥，蒸发（用纱布干燥常常引入纤维污染物是很难除去）。体视显微镜下指导（1 - 3倍），并用干净的艺术家刷（10-0驼绒），适用于环氧 - 酚醛粘合剂的薄膜，以一个元件的背面，并立即放置在接触相对的背面的第二元件的并对准箔网格（ 图1A）。 </li><li>将在50粘合元素 - 60℃的烘箱中的O / N至完全固化的环氧树脂。 注意:不要用镊子保税元素，因为过多的环氧树脂可渗入到元件和压力可能会造成电网的错位。该两部分环氧具有混合后可用冷藏寿命只有6个星期。债券和治疗元素的充足供应一次，并将它们存储在一个干净，无灰尘，环境，供以后使用。 </li></ol></li><li>选型和布线双晶应变计<ol><li>修剪结合双元素的3×3mm的最终大小与＃11手术刀或单刃刀片。延迟修剪此时的双元件的最高部分，以便有一个区域，用于安全地处理元件（ 图1A）。 </li><li>每个元件都需要从三芯，粘合制成的四芯线，铁氟龙绝缘电线（P / N:336-FTE）。拆卸的三芯线之一30厘米编织股线分成三个构成线。 </li><li>为了让一个四线电缆，配对所产生的单线与包含三芯线的二三十厘米长的范围内相同颜色的线之一。在下面的步骤中，这些匹配的彩色导线会在末端被接合以形成一个共同的电线进行最后的应变仪（ 图1B）。 </li><li>从每根电线与热剥线机的两端除去1mm左右聚四氟乙烯绝缘的。采用活性松香溶胶dering通量和低温焊料（熔点183℃）用锡焊的铅笔线结束。 </li><li>在接下来的阶段中，microsoldering尖端仍需以形成多个离散的焊点，以防止热损坏的元件（ 图1C）的薄膜层。为了制造更小的microsoldering前端，包住一小块铜丝（〜0.250毫米直径）绕一圈的标准焊头，确保铜导线延伸超出标准焊头的长度。 </li><li>磁通只在粘接的双元件的一侧的焊剂焊盘用清洁刷10-0和焊料单一导线和一对公共引线的焊接垫（ 图1D）之一。剩余磁通之后可以用干净毛刷蘸树脂的溶剂清除。 </li><li>重复的相反侧的过程中，以确保剩余的公共引线焊接到焊盘相对原来的共用导线。 </li></ol></li><li>测试第二epoxying双元件应变计<ol><li>焊接金插座连接器（E363 / 0）到引线的自由端。在这一点上，应变计连接到一记录放大器（在下面描述），以测试双元件组件的完整性。 </li><li>衡量一个良好的质量电压电阻计的电阻。元素登记约350Ω的电阻。在这一点上与新鲜焊料重焊不充分的连接。 </li><li>如果焊接连接和双素体被认为是令人满意的，修剪掉任何剩余的元素影片。 </li><li>绝缘上的元件的焊盘的焊点有一层薄薄的两部分组成的，用于硅橡胶的环氧树脂（P / N:E211）。为获得最佳效果，部分地固化该树脂为20 - 30分钟之前，应用（ 图1E）。 </li></ol></li><li>封装在硅双晶应变计<ol><li>切成三片0.5毫米厚的硅树脂片材（P / N:20-20）到15mm 2和c瘦用蒸馏水硅氧烷。切割一块硅薄片的成U字形，以便适应最终的双元件装配而不会变形包封硅氧烷（ 图1F）。 </li><li>涂层的缺口无硅酮片与透明的硅粘合剂的内表面上。 </li><li>夹在中间的双元件组件的槽口和对准的外板内，然后轻轻地压出任何过量的有机硅，以及气泡，从中心向外。小心夹住的金属棒材两个块之间的密封组件，放置24小时，以确保均匀的厚度，并且没有变形发生。 </li><li>允许过量的硅保持沿装配和固化的边界。当纸张硅酮修剪成所需的最终尺寸（; 图1G常用6毫米×8 mm）该过量将被删除。 </li></ol></li><li>导线连接器和校准完成<ol><li>加强焊点对个人端子线黄金插座连接器的引线，3毫米（1/8英寸），热 ​​缩管，并在一个塑料电极座（MS363， 图1H）对齐。固定电极底座和电线0.125和0.25英寸直径的热缩管，以防止实验（ 图1I）在断开连接。 </li><li>应变计信号通过一个高增益放大器，桥接（P / N:AMP-01-SG）的处理。连接用电缆配套插头应变计到放大器（363 - SL / 6）来匹配电极座。螺纹瓶盖提供了额外的安全性，在实验过程中保持不间断的信号。 </li><li>调整放大器上的桥，平衡和增益设置，按制造商的指示专门的应变计。粘贴应变片的结尾，其中的电线横退出，以坚硬夹和放置在另一端一条1G的静载荷校准最初由Pascaud和他的同事描述2，16，17。 </li></ol></li></ol> 2，外科手术应变计急性植入<ol><li>动物护理和准备工作: <ol><li>夜食物剥夺试验动物手术植入前（水可随意提供）。 </li><li>深深地麻醉的动物。 Thiobutabarbital（100 - 150毫克/公斤; ip进行大鼠），优选为终端应变植入和实验，由于在大鼠10持续的麻醉效果和胃的反射最小的改动（ 即 ，非存活）。测试没有爪子捏反射来判断麻醉深度。 </li><li>准备大鼠无菌手术的复杂程度取决于实验设计和批准IACUC准则，包括消毒外科手术刀具，刮脸切口部位，应用审核眼膏和消毒所有的外科手术领域。 注意:如果应变计适于长期使用，消毒应变计只与环氧乙烷（气体）灭菌。利用热或化学灭菌技术可能会损坏应变计。 </li></ol></li><li>气管插管终端的实验: <ol><li>对于持续时间长，末端，实验插管的大鼠气管套管，以保持气道畅通。使1 - 从下颌到胸骨切迹下缘的颈部腹侧2厘米正中切口。 </li><li>单独使用钝性分离，在正中线，显露气管底层肩带肌肉。从底层隔离食道气管和地方的气管和食管之间3-0 ethilon缝合循环，形成一个连字。 </li><li>通过一个小incison在膜的两个气管软骨环只是远离甲状腺之间向前方打开气管。将一小片的聚乙烯管（P / N:PE-270），在5mm的长度（和斜角的一端）的进气管并固定到位结扎。 </li><li>把带状肌放回原处，并缝合皮肤覆以3-0 ethilon。 </li></ol></li><li>应变计仪器仪表胃肠道面: <ol><li>主题四个角应变计与4 - 4-0使用在手术前＃14锥度点3/8圆针5厘米长度，或更小的，无菌的丝线。丝缝合线提供的灵活性高的水平，并且更不容易损坏的硅氧烷密封应变计元件。 注:丝线是可以接受的非手术生存和内部应用程序，其中的细菌穿过上皮屏障排汗不是一个风险。在需要存活手术应用中，Prolene线缝合是必要的，以减少感染所固有的丝缝合线的编织布料的纤维的风险。 </li><li>通过切开沿中线腹部皮肤进行剖腹手术。部分腹直肌肌沿连接白线（缺血性），以防止出血。然后做一个非常肤浅的正中切口，在腹膜壁层，以免划破底层腹腔脏器。 </li><li> Exteriorize用生理盐水浸湿棉签的帮助胃。保持胃的位置小心地将它放在盐水浸泡纱布垫在腹部切口的尾端。 </li><li>对准封装应变平行于环形平滑肌纤维的网格。使用先前螺纹缝线（步骤2.3.1），附加计的角落，用＃14锥度点3/8圆针胃体的腹侧浆膜面。为了减少组织损伤和潜在的出血，不使用切削刃的针和不穿孔任何浅表血管的胃的表面上。 </li><li>开始计的缝线图案沿着附近的眼底/语料库边界胃的大弯，然后继续下沿眼底/胼边界朝向小弯。应变计底层浆膜应该既不是松弛也不过分拉伸，以便获得最好的结果。 </li><li>小心用盐水浸泡过的棉签肚子返回到它的解剖位置。 </li><li>在急性模型，exteriorize应变计导致的正中切口的腹壁切口闭合前的尾端。为了操纵该动物或端线连接器的过程中提供应变消除固定的自由电线的动物（ 例如 ，后足）。单独关闭腹直肌肌和腹部皮肤用3-0尼龙线。在慢性模型，皮下固定引线沿大鼠背侧和exteriorize他们的头骨18以上。 </li><li>手术器械后，将动物在立体定位框架，用于支撑头部和提高上部躯干。后者步骤有助于降低再录制过程中抽出鼻唇神器。监测肛温，并在37±1°C的使用反馈控制的加热垫维持。 </li><li>在利用thiobutabarbital麻醉终端实验结束时，动物必须被安乐死与安乐死美国兽医协会（AVMA）指南的方式。 </li></ol></li><li>胃动力录音: <ol><li>放大用任何市售的直流桥式放大器的应变计信号。 </li><li>记录使用任何市售的数据采集系统的图表记录功能的计算机上的DC输出信号。 注:放大器输出的硬拷贝可以通过测谎图表记录产生。 </li></ol></li></ol>胃收缩代表性的测量跟踪脑干刺激<ol><li>曝光脑干及第四脑室<ol><li>手术器械和placem后在立体框架的Thiobutabarbital麻醉动物耳鼻喉科，做一个1.5 - 2厘米中线切开皮肤，从枕骨向颈部的基地。 </li><li>分离结缔组织沿中线加盟底层颈部肌肉的双侧腹部肌肉（肌肉从浅到深的提耳长肌颅部，提耳长肌尾部分，和颈阔肌颅部分）。 </li><li>从枕骨分离提耳后长肌的中线是明确和公开。 </li><li>小心使用钝性分离分离从根本硬膜颈阔肌暴露颅骨的尾部区域。 </li><li>使用新的地下25针仔细分离跟着枕髁两侧延伸枕骨大孔硬膜。 </li><li>使用＃5杜蒙钳去除PIA和脑膜蛛网膜覆第四脑室和揭露脑干。 </li></ol></li><li>管理第四脑室​​促甲状腺激素释放激素或静脉注射硝普钠<ol><li>称重和溶解促甲状腺激素释放激素（TRH），在无菌盐水中，达到50μM的TRH的最终浓度。 </li><li>称重和溶解硝普钠（SNP）中的无菌盐水以达到150μM的SNP的最终浓度。 </li><li>用10微升注射器，辖2微升的TRH（最终剂量等于100皮摩尔）脑干第四脑室的背水面，以方便记录胃收缩。 </li><li>使用无菌注射器和27G的针头，辖150微摩尔/千克的SNP通过尾静脉，以方便记录胃放松。 </li></ol></li></ol>

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Liver Physiol. 290 (1), G74-G82 (2005).</li><li>Bass, P., Wiley, J. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contractile+force+transducer+for+recording+muscle+activity+in+unanesthetized+animals.">Contractile force transducer for recording muscle activity in unanesthetized animals.</a> J. Appl. Physiol. 32 (4), 567-570 (1972).</li><li>Holmes, G. M., Browning, K. N., Tong, M., Qualls-Creekmore, E., Travagli, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vagally+mediated+effects+of+glucagon-like+peptide+1:+in+vitro+and+in+vivo+gastric+actions.">Vagally mediated effects of glucagon-like peptide 1: in vitro and in vivo gastric actions.</a> J. Physiol. 587 (19), 4749-4759 (2009).</li><li>Tong, M., Qualls-Creekmore, E., Browning, K. N., Travagli, R. A., Holmes, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+spinal+cord+injury+in+rats+diminishes+vagally-mediated+gastric+responses+to+cholecystokinin-8s.">Experimental spinal cord injury in rats diminishes vagally-mediated gastric responses to cholecystokinin-8s.</a> Neurogastroenterol. Motil. 23 (2), e69-e79 (2011).</li><li>Miyano, 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=The+role+of+the+vagus+nerve+in+the+migrating+motor+complex+and+ghrelin-+and+motilin-induced+gastric+contraction+in+suncus.">The role of the vagus nerve in the migrating motor complex and ghrelin- and motilin-induced gastric contraction in suncus.</a> PLoS ONE. 8 (5), e64777 (2013).</li><li>Holmes, G. M., Rogers, R. C., Bresnahan, J. C., Beattie, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thyrotropin-releasing+hormone+(TRH)+and+CNS+regulation+of+anorectal+motility+in+the+rat.">Thyrotropin-releasing hormone (TRH) and CNS regulation of anorectal motility in the rat.</a> J Auton. Nerv. Syst. 56, 8-14 (1995).</li><li>Ormsbee, H. S., Bass, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gastroduodenal+motor+gradients+in+the+dog+after+pyloroplasty.">Gastroduodenal motor gradients in the dog after pyloroplasty.</a> Am. J. Physiol. 230, 389-397 (1976).</li><li>Fukuda, H., 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=Impaired+gastric+motor+activity+after+abdominal+surgery+in+rats.">Impaired gastric motor activity after abdominal surgery in rats.</a> Neurogastroenterol. Motil. 17 (2), 245-250 (2005).</li><li>Browning, K. N., Babic, T., Holmes, G. M., Swartz, E., Travagli, R. 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+critical+re-evaluation+of+the+specificity+of+action+of+perivagal+capsaicin.">A critical re-evaluation of the specificity of action of perivagal capsaicin.</a> J. Physiol. 591 (6), 1563-1580 (2013).</li></ol>

The authors declare that they have no competing financial interests. The suppliers listed in this manuscript are provided for reference only.

这里介绍的方法允许单个实验室制造灵敏微型应变计用于生物应用，包括但不限于，胃肠道蠕动在小的实验室动物。由于这些应变片的商业化生产已经停止，实验室研究胃肠功能仅限于可能不允许全方位的实验应用可用的其他技术。这个报告对先前所描述的技术15的更新和更详细的描述。文本和相应的视频专门解决方案，使我们的发展制造工艺的掌握和认识过程中常见的陷阱。 <p class="...

在体内胃肠道（GI）在多个实验条件蠕动的记录仍然是一个强大的工具，了解必要的营养平衡的基本正常和病理生理过程的实验研究。传统上，无数的实验方法，用一些相似的临床实践1，已经采用直接量化的改变胃肠道的收缩率2-5，腔内压力6，7，或不可吸收标志物8的胃肠道转运，发现9或稳定同位素10-12。每种技术都有其独特的优点和缺点，这在文献中先前已经解决。例如，气囊测压的效用量化的压力变化受到质疑由于气囊材料的固有遵守而不可吸收的标记物胃肠恢复需要安乐死试验阴性特质l对于单个数据点。最近，一个微型动脉压导管的应用和验证有报道称，提供了在大鼠和小鼠3监测胃收缩的非手术方法。而一个orogastrically放置压力传感器有效地避免侵入性手术过程中消除了对胃肠功能混杂变量，这种方法只适用于麻醉制剂。此外，由于缺乏视觉引导不允许胃的特定区域内的换能器的相一致的位置。因此，本申请中限定为胃或结肠因为可视化，再加上相对较硬的传感器导线，十二指肠内或回肠不是一个选项。 同样，生物磁交流电biosusceptometry（ACB）技术已被证实为胃肠道收缩分析4。而ACB技术提供了一种非侵入性的APproach测定胃肠道收缩，ACB患有中，类似的限制，利用摄入磁检测介质中，没有允许在胃肠道的特定区域的精确记录。这一限制可以通过对磁性标记的手术植入来克服。尽管如此，ACB技术的必要条件是该动物被麻醉，进行数据收集。 Ultrasonomicrometry已经用于一些GI，以便采取尺寸小，空间的优势研究13，第14，和压电晶体发射器/接收器的时间的优点。胃平滑肌收缩波都没有的高频事件和发生在大约3的速率 - 5个周期/分钟。因此，声纳微测量的时间的优点可能是不必要的，以证明费用。此外，尽管线性运动被准确地测量与声纳微测量，限制已经提出关于准确胃肠数据解释可能导致植入晶体14的数量不足的。 基于Bass和同事2,15的原创设计该协议的可视化更充分地记载了一步一步的制造和试验应用缩影，双晶应变计是具有高灵敏度和灵活性，在整个胃肠记录平滑肌收缩道。应变计元件的尺寸适合于任何啮齿类动物的应用，因为灵敏度成品应变计和尺寸是最依赖于硅薄片封装的元件。这些应变片很容易适应的麻醉和自由活动的实验动物模型急性和慢性的应用程序，从而提供量化平滑肌收缩单一技术。

<table><tbody><tr><td>Strain gage element</td><td>Micro-Measurements (Vishay Product Group)</td><td>EA-06-031-350</td><td>&amp;nbsp;Linear pattern, foil, stress analysis strain gage (2 required)&lt;br/&gt; &lt;a href=&quot;http://www.vishaypg.com/micro-measurements/&quot;&gt;www.vishaypg.com/micro-measurements/&lt;/a&gt;&lt;br/&gt; or&lt;br/&gt; &lt;a href=&quot;http://www.vishaypg.com/docs/11070/031ce.pdf&quot;&gt;http://www.vishaypg.com/docs/11070/031ce.pdf&lt;/a&gt;</td></tr><tr><td>Epoxy-phenolic adhesive</td><td>M-bond 610</td><td>General purpose adhesive for bonding strain gage elements</td><td>&lt;a href=&quot;http://www.vishaypg.com/docs/11024/wirecable.pdf&quot;&gt;http://www.vishaypg.com/docs/11024/wirecable.pdf&lt;/a&gt;</td></tr><tr><td>3 conductor insulated wire</td><td>336-FTE</td><td>Fine gage, flexible general purpose wire</td><td>&lt;a href=&quot;http://www.vishaypg.com/docs/11024/wirecable.pdf&quot;&gt;http://www.vishaypg.com/docs/11024/wirecable.pdf&lt;/a&gt;</td></tr><tr><td>Flux and rosin solvent kit</td><td>FAR-2 M-Flux AR kit</td><td>Liquid solder flux</td><td>&lt;a href=&quot;http://www.vishaypg.com/docs/11023/soldacce.pdf&quot;&gt;http://www.vishaypg.com/docs/11023/soldacce.pdf&lt;/a&gt;</td></tr><tr><td>Solder</td><td>361A-20R-25</td><td>Optimized and recommended for strain gage applications</td><td>&lt;a href=&quot;http://www.vishaypg.com/docs/11023/soldacce.pdf&quot;&gt;http://www.vishaypg.com/docs/11023/soldacce.pdf&lt;/a&gt;</td></tr><tr><td>Gold socket connector</td><td>PlasticsOne</td><td>E363/0</td><td>Socket contact for electrode pedestal&lt;br/&gt; &lt;a href=&quot;http://www.plastics1.com/PCR/Catalog/Item.php?item=407&quot;&gt;http://www.plastics1.com/PCR/Catalog/Item.php?item=407&lt;/a&gt;</td></tr><tr><td>Electrode pedestal</td><td>MS363</td><td>Secure platform for wire contacts</td><td>&lt;a href=&quot;http://www.plastics1.com/PCR/Catalog/Item.php?item=499&quot;&gt;http://www.plastics1.com/PCR/Catalog/Item.php?item=499&lt;/a&gt;</td></tr><tr><td>6-wire cable</td><td>363 PLUG W/VINYL SL/6</td><td>Pre-fabricated vinyl-coated cable (in customized lengths) with plug adaptor to match electrode pedestal and tinned solder lugs on terminal end</td><td></td></tr><tr><td>Silicone rubber casting compound</td><td>EIS electrical products</td><td>Elan Tron E211</td><td>Potting medium for gage/wire solder joints&lt;br/&gt; &lt;a href=&quot;http://www.eis-inc.com/&quot;&gt;http://www.eis-inc.com&lt;/a&gt;</td></tr><tr><td>HOTweezers</td><td>Meisei Corporation</td><td>Model 4B</td><td>Wire insulation strippers&lt;br/&gt; &lt;a href=&quot;http://www.impexron.us/&quot;&gt;http://www.impexron.us&lt;/a&gt;</td></tr><tr><td>Soldering station</td><td>Weller (Apex Tool Group)</td><td>WES 51</td><td>High quality soldering equipment&lt;br/&gt; &lt;a href=&quot;http://www.apexhandtools.com/weller/index.cfm&quot;&gt;http://www.apexhandtools.com/weller/index.cfm&lt;/a&gt;</td></tr><tr><td>Available through http://www.eis-inc.com or http://www.amazon.com</td><td></td><td></td><td></td></tr><tr><td>Silicone sheet</td><td>Trelleborg Sealing Solutions Northborough-Life Sciences</td><td>Pharmelast 20-20</td><td>Encapsulating strain gage elements&lt;br/&gt; 10 B Forbes Road Northborough, MA 01532 (800) 634-2000</td></tr><tr><td>Amplifier</td><td>Experimetria Ltd</td><td>AMP-01-SG</td><td>&lt;br/&gt; &lt;a href=&quot;http://experimetria.com/Biological_amplifiers.php&quot;&gt;http://experimetria.com/Biological_amplifiers.php&lt;/a&gt;</td></tr></tbody></table>

fabrication and implantation of miniature dual-element strain gages for measuring in vivo gastrointestinal contractions in rodents.

Gastrointestinal dysfunction remains a major cause of morbidity and mortality. Indeed, gastrointestinal (GI) motility in health and disease remains an area of productive research with over 1,400 published animal studies in just the last 5 years. Numerous techniques have been developed for quantifying smooth muscle activity of the stomach, small intestine, and colon. In vitro and ex vivo techniques offer powerful tools for mechanistic studies of GI function, but outside the context of the integrated systems inherent to an intact organism. Typically, measuring in vivo smooth muscle contractions of the stomach has involved an anesthetized preparation coupled with the introduction of a surgically placed pressure sensor, a static pressure load such as a mildly inflated balloon or by distending the stomach with fluid under barostatically-controlled feedback. Yet many of these approaches present unique disadvantages regarding both the interpretation of results as well as applicability for in vivo use in conscious experimental animal models. The use of dual element strain gages that have been affixed to the serosal surface of the GI tract has offered numerous experimental advantages, which may continue to outweigh the disadvantages. Since these gages are not commercially available, this video presentation provides a detailed, step-by-step guide to the fabrication of the current design of these gages. The strain gage described in this protocol is a design for recording gastric motility in rats. This design has been modified for recording smooth muscle activity along the entire GI tract and requires only subtle variation in the overall fabrication. Representative data from the entire GI tract are included as well as discussion of analysis methods, data interpretation and presentation.

从Thiobutabarbital-麻醉大鼠的代表性数据示于图2中的顶部曲线表示的促甲状腺激素释放激素脑干给药（TRH，100皮摩尔），一种已知的能动性增强肽3,19期间从大鼠胃体收缩。它示出了现有的增加，相位胃平滑肌活性的基线收缩。注:在胃的收缩这些峰分析跟踪方法由安庶庇和低音20原配方运动指数=（N + 1×1）+（N 2×2）+（N 3×4）+（N 4个 x8） <...

Watch this Scientific Journal Video about Fabrication and Implantation of Miniature Dual-element Strain Gages for Measuring In Vivo Gastrointestinal Contractions in Rodents. at JoVE.com

Fabrication and Implantation of Miniature Dual-element Strain Gages for Measuring In Vivo Gastrointestinal Contractions in Rodents.

The in vivo measurement of smooth muscle contractions along the gastrointestinal tract of laboratory animals remains a powerful, though underutilized, technique. Flexible, dual element strain gages are not commercially available and require fabrication. This protocol describes the construction of reliable, inexpensive strain gages for acute or chronic implantation in rodents.

的微型双元应变计测量加工和植入<em&gt;体内</em&gt;胃肠收缩鼠害。

Gastrointestinal dysfunction remains a major cause of morbidity and mortality. Indeed, gastrointestinal (GI) motility in health and disease remains an ...

fabrication-implantation-miniature-dual-element-strain-gages-for

Universidad de Cartagena UDEC

Research

JoVE Journal

Bioengineering

Fabrication and Implantation of Miniature Dual-element Strain Gages for Measuring In Vivo Gastrointestinal Contractions in Rodents.

10.3K 次观看.  Penn State University College of Medicine. The in vivo measurement of smooth muscle contractions along the gastrointestinal tract of laboratory animals remains a powerful, though underutilized, technique. Flexible, dual element strain gages are not commercially available and require fabrication. This protocol describes the construction of reliable, inexpensive strain gages for acute or chronic implantation in rodents.

视频: Fabrication and Implantation of Miniature Dual-element Strain Gages for Measuring In Vivo Gastrointestinal Contractions in Rodents.

An in vivo Rodent Model of Contraction-induced Injury and Non-invasive Monitoring of Recovery

Muscle strains are one of the most common complaints treated by physicians. A muscle injury is typically diagnosed from the patient history and physical exam alone, however the clinical presentation can vary greatly depending on the extent of injury, the patient's pain tolerance, etc. In patients with muscle injury or muscle disease, assessment of muscle damage is typically limited to clinical signs, such as tenderness, strength, range of motion, and more recently, imaging studies. Biological markers, such as serum creatine kinase levels, are typically elevated with muscle injury, but their levels do not always correlate with the loss of force production. This is even true of histological findings from animals, which provide a "direct measure" of damage, but do not account for all the loss of function. Some have argued that the most comprehensive measure of the overall health of the muscle in contractile force. Because muscle injury is a random event that occurs under a variety of biomechanical conditions, it is difficult to study. Here, we describe an in vivo animal model to measure torque and to produce a reliable muscle injury. We also describe our model for measurement of force from an isolated muscle in situ. Furthermore, we describe our small animal MRI procedure.

An in vivo Rodent Model of Contraction-induced Injury and Non-invasive Monitoring of Recovery

An in vivo animal model of injury is described. The method takes advantage of the subcutaneous position of the fibular nerve. Velocity, timing of muscle activation, and arc of motion are all pre-determined and synchronized using commercial software. Post injury changes are monitored in vivo using MR imaging/spectroscopy.

一个在体内收缩引起的损伤，非侵入性的监测恢复啮齿动物模型

Muscle strains are one of the most  common complaints treated by physicians. A muscle injury is typically diagnosed from the patient history and  physical ...

科研

医学

Physiological Experimentation with the Crayfish Hindgut: A Student Laboratory Exercise

The purpose of the report is to describe dissection techniques for preparing the crayfish hindgut and to demonstrate how to make physiological recordings with a force transducer to monitor the strength of contraction. In addition, we demonstrate how to visually monitor peristaltic activity, which can be used as a bioassay for various peptides, biogenic amines and neurotransmitters. This preparation is amenable to student laboratories in physiology and for demonstrating pharmacological concepts to students. This preparation has been in use for over 100 years, and it still offers much as a model for investigating the generation and regulation of peristaltic rhythms and for describing the mechanisms underlying their modulation. The pharmacological assays and receptor sub-typing that were started over 50 years ago on the hindgut still contribute to research today. This robust preparation is well suited to training students in physiology and pharmacology.

In this report we demonstrate techniques that can be used to investigate the biology of the crayfish hindgut. We show how to dissect a crayfish abdomen and study the associated anatomy, physiology and modulation of activity. The peristaltic activity and strength of contractions are measured using a force transducer.

生理实验与小龙虾肠：学生实验室的工作

The purpose of the report is to describe dissection techniques for preparing the crayfish hindgut and to demonstrate how to make physiological recordings ...

神经科学

Intrauterine Telemetry to Measure Mouse Contractile Pressure In Vivo

A complex integration of molecular and electrical signals is needed to transform a quiescent uterus into a contractile organ at the end of pregnancy. Despite the discovery of key regulators of uterine contractility, this process is still not fully understood. Transgenic mice provide an ideal model in which to study parturition. Previously, the only method to study uterine contractility in the mouse was ex vivo isometric tension recordings, which are suboptimal for several reasons. The uterus must be removed from its physiological environment, a limited time course of investigation is possible, and the mice must be sacrificed. The recent development of radiometric telemetry has allowed for longitudinal, real-time measurements of in vivo intrauterine pressure in mice. Here, the implantation of an intrauterine telemeter to measure pressure changes in the mouse uterus from mid-pregnancy until delivery is described. By comparing differences in pressures between wild type and transgenic mice, the physiological impact of a gene of interest can be elucidated. This technique should expedite the development of therapeutics used to treat myometrial disorders during pregnancy, including preterm labor.

Intrauterine Telemetry to Measure Mouse Contractile Pressure In Vivo

This manuscript provides a protocol for implanting telemeters in the mouse for the purpose of measuring intrauterine pressures during pregnancy.

宫内遥测来衡量鼠标收缩压力<em&gt;在体内</em

A complex integration of molecular and electrical signals is needed to transform a quiescent uterus into a contractile organ at the end of pregnancy. ...

Wireless Intramuscular EMG System for Locomotion Studies in Stroke Model Rats

A Real-Time Wearable Electromyography Measurement System for Small Animals

The intramuscular electromyography (EMG) measurement method for experimental animals has been implemented in various ways. Among these methods, tethering cables to external measurement devices can restrict the movement of experimental animals, while implantable devices may cause unwanted side effects due to the constant presence of a device with considerable size and weight. To address these issues, we propose a low-cost, wireless, detachable EMG measurement system and experimental procedure. This article focuses on the surgical installation of intramuscular wire electrodes with small connectors and the development of the wireless system. Notably, in this system, only the wire electrodes are inserted into the animal's body. Using this system, EMG measurements can be easily performed by attaching the circuit system to a connector installed on the animal's back, with real-time monitoring achievable on a laptop. The proposed method is explained in a detailed, step-by-step manner, followed by a demonstration involving the insertion of intramuscular electrodes into the hindlimbs of a rat. A treadmill experiment is conducted for a locomotion study, and the resulting electrophysiological signals are subsequently obtained and analyzed.

A protocol is presented for the installation and preparation of intramuscular electrodes, along with an animal-mountable, miniaturized measurement device for electromyography (EMG) analysis in locomotion studies. This system enables the wireless transmission of real-time EMG data for investigating locomotion recovery in stroke model rats.

The intramuscular electromyography (EMG) measurement method for experimental animals has been implemented in various ways. Among these methods, tethering ...

工程学

Gastrointestinal Motility Monitor (GIMM)

The Gastrointestinal Motility Monitor (GIMM; Catamount Research and Development; St. Albans, VT) is an in vitro system that monitors propulsive motility in isolated segments of guinea pig distal colon. The complete system consists of a computer, video camera, illuminated organ bath, peristaltic and heated water bath circulating pumps, and custom GIMM software to record and analyze data. Compared with traditional methods of monitoring colonic peristalsis, the GIMM system allows for continuous, quantitative evaluation of motility. The guinea pig distal colon is bathed in warmed, oxygenated Krebs solution, and fecal pellets inserted in the oral end are propelled along the segment of colon at a rate of about 2 mm/sec. Movies of the fecal pellet proceeding along the segment are captured, and the GIMM software can be used track the progress of the fecal pellet. Rates of propulsive motility can be obtained for the entire segment or for any particular region of interest. In addition to analysis of bolus-induced motility patterns, spatiotemporal maps can be constructed from captured video segments to assess spontaneous motor activity patterns. Applications of this system include pharmacological evaluation of the effects of receptor agonists and antagonists on propulsive motility, as well as assessment of changes that result from pathophysiological conditions, such as inflammation or stress. The guinea pig distal colon propulsive motility assay, using the GIMM system, is straightforward and simple to learn, and it provides a reliable and reproducible method of assessing propulsive motility.

Gastrointestinal Motility Monitor GIMM

Evaluation of colonic motility in the guinea pig distal colon with the Gastrointestinal Motility Monitor (GIMM) is a straightforward and simple to learn approach to quantitatively evaluate propulsive motility in the gastrointestinal tract.

肠胃蠕动监视器（GIMM）

The Gastrointestinal Motility Monitor (GIMM; Catamount Research and Development; St. Albans, VT) is an in vitro system that monitors propulsive motility ...

生物学

Using a Microfluidics Device for Mechanical Stimulation and High Resolution Imaging of C. elegans

One central goal of mechanobiology is to understand the reciprocal effect of mechanical stress on proteins and cells. Despite its importance, the influence of mechanical stress on cellular function is still poorly understood. In part, this knowledge gap exists because few tools enable simultaneous deformation of tissue and cells, imaging of cellular activity in live animals, and efficient restriction of motility in otherwise highly mobile model organisms, such as the nematode Caenorhabditis elegans. The small size of C. elegans makes them an excellent match to microfluidics-based research devices, and solutions for immobilization have been presented using microfluidic devices. Although these devices allow for high-resolution imaging, the animal is fully encased in polydimethylsiloxane (PDMS) and glass, limiting physical access for delivery of mechanical force or electrophysiological recordings. Recently, we created a device that integrates pneumatic actuators with a trapping design that is compatible with high-resolution fluorescence microscopy. The actuation channel is separated from the worm-trapping channel by a thin PDMS diaphragm. This diaphragm is deflected into the side of a worm by applying pressure from an external source. The device can target individual mechanosensitive neurons. The activation of these neurons is imaged at high-resolution with genetically-encoded calcium indicators. This article presents the general method using C. elegans strains expressing calcium-sensitive activity indicator (GCaMP6s) in their touch receptor neurons (TRNs). The method, however, is not limited to TRNs nor to calcium sensors as a probe, but can be expanded to other mechanically-sensitive cells or sensors.

Using a Microfluidics Device for Mechanical Stimulation and High Resolution Imaging of C. elegans

New tools for mechanobiology research are needed to understand how mechanical stress activates biochemical pathways and elicits biological responses. Here, we showcase a new method for selective mechanical stimulation of immobilized animals with a microfluidic trap allowing high-resolution imaging of cellular responses.

利用微流体装置进行机械刺激和高分辨率成像的研究C.线虫

One central goal of mechanobiology is to understand the reciprocal effect of mechanical stress on proteins and cells. Despite its importance, the ...

Studying Murine Small Bowel Mechanosensing of Luminal Particulates

Gastrointestinal (GI) motility is critical for normal digestion and absorption. In the small bowel, which absorbs nutrients, motility optimizes digestion and absorption. For this reason, some of the motility patterns in the small bowel include segmentation for mixing of luminal contents and peristalsis for their propulsion. Physical properties of luminal contents modulate the patterns of small bowel motility. The mechanical stimulation of GI mechanosensory circuits by transiting luminal contents and underlying gut motility initiate and modulate complex GI motor patterns. Yet, the mechanosensory mechanisms that drive this process remain poorly understood. This is primarily due to a lack of tools to dissect how the small bowel handles materials of different physical properties. To study how the small bowel handles particulates of varying sizes, we have modified an established in vivo method to determine small bowel transit. We gavage live mice with fluorescent liquid or tiny fluorescent beads. After 30 minutes, we dissect out the bowels to image the distribution of fluorescent contents across the entirety of the GI tract. In addition to high-resolution measurements of the geometric center, we use variable size binning and spectral analysis to determine how different materials affect small bowel transit. We have explored how a recently discovered "gut touch" mechanism affects small bowel motility using this approach.

To study how the small bowel handles particulates of varying sizes, we have modified an established in vivo method to determine small bowel transit.

研究小鼠小肠对管腔颗粒的机械传感

Gastrointestinal (GI) motility is critical for normal digestion and absorption. In the small bowel, which absorbs nutrients, motility optimizes digestion ...

<meta charset="utf8"></head><body>通过小鼠经颅钙成像研究肠脑相互作用

Real-time Analysis of Gut-brain Neural Communication: Cortex wide Calcium Dynamics in Response to Intestinal Glucose Stimulation

Communication between the gastrointestinal tract and the brain after nutrient absorption plays an essential role in food preference, metabolism, and feeding behaviors. Particularly concerning specific nutrients, many studies have elucidated that the assimilation of glucose within gut epithelial cells instigates the activation of many signaling molecules. Hormones such as glucagon-like peptide-1 are renowned as quintessential signaling mediators. Since hormones predominantly influence the brain through circulatory pathways, they slowly modulate brain activity.
However, recent studies have shown two expeditious gut-brain pathways facilitated by the autonomic nervous system. One operates via the spinal afferent neural pathway, while the vagus nerve mediates the other. Consequently, brain responses following glucose assimilation in the gastrointestinal tract are complicated. Moreover, as intestinal stimulation finally induces diverse cortical activities, including sensory, nociceptive, reward, and motor responses, it is necessary to employ methodologies that facilitate the visualization of localized brain circuits and pan-cortical activities to comprehend gut-brain neural transmission fully. Some studies have indicated precipitous alterations in calcium ion (Ca2+) concentrations within the hypothalamus and ventral tegmental area independently through different pathways after intestinal stimulation. However, whether there are changes in cerebral cortex activity has not been known.
To observe cerebral cortex activity after intragastric glucose injection, we developed an imaging technique for real-time visualization of cortex wide Ca2+ dynamics through a fully intact skull, using transgenic mice expressing genetically encoded Ca2+ indicators. This study presents a comprehensive protocol for a technique designed to monitor intestinal stimulation-induced transcranial cortex wide Ca2+ imaging following intragastric glucose injection via an implanted catheter. The preliminary data suggest that administering glucose solution into the gut activates the frontal cortex, which remains unresponsive to water administration.

<meta charset="utf8"></head><body>肠脑神经通讯实时分析:响应肠道葡萄糖刺激的皮层宽钙动力学

Gut-brain communication, facilitated by the vagus nerve, is crucial for communication between the gastrointestinal endocrine system and the brain. However, whether intragastric glucose injection can change cortical activity is still not understood. Here, we offer a comprehensive protocol to observe changes in cortical activity after glucose injection into the duodenum.

肠脑神经通讯实时分析:响应肠道葡萄糖刺激的皮层宽钙动力学

Communication between the gastrointestinal tract and the brain after nutrient absorption plays an essential role in food preference, metabolism, and ...

的微型双元应变计测量加工和植入

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研究小鼠小肠对管腔颗粒的机械传感

肠脑神经通讯实时分析:响应肠道葡萄糖刺激的皮层宽钙动力学

肠脑神经通讯实时分析:响应肠道葡萄糖刺激的皮层宽钙动力学

肠脑神经通讯实时分析:响应肠道葡萄糖刺激的皮层宽钙动力学