这项工作是从宾夕法尼亚州卫生署的拨款支持。 米格尔加西亚托斯卡诺博士是坦普尔大学的​​博士后研究员，在这项研究中的阿方马丁Escudero基​​金会资助。 我们要感谢的视频制作Iliya Yordanov和凯文Kotredes。

1。作为人类脓毒症小鼠模型的盲肠结扎穿孔对于此过程的C57BL / 6小鼠（7-9周龄）。 <ol><li> 1:1氯胺酮（75mg/kg）和甲苯噻嗪（按15mg/kg）的解决方案由腹腔注射麻醉鼠标。作为参考，一个重达20克的小鼠注入30μL1:1的解决方案。另外，可以使用麻醉蒸发器吸入异氟醚麻醉的小鼠。 </li><li>剃须鼠标腹部和消毒面积首次申请优碘用70％酒精棉签（1）在下面的图片序列擦拭的解决方案。洗必泰可作为替代消毒剂。可选：无菌的悬垂性，可用于维护该地区的清洁。 </li><li>在无菌条件下，实行1至2厘米的中线剖腹探查术和揭露与毗邻大肠盲肠（2和3）。 </li><li> 6.0丝线缝合（6-0普理灵，8680G;爱惜康）在其下面的回肠盲肠的阀门基地，是紧紧盲肠结扎穿孔一次或两次用19号针头（4和5）在同一一侧盲肠。请注意结扎盲肠结扎点从盲肠末端的距离定义的长度，将决定其严重程度。距离&gt;1厘米生产高档败血症≤1厘米的距离而产生中旬低档败血症。另外请注意，使用方法，这里显示的盲肠穿孔通过，并通过穿刺技术（通过盲肠针）的标准是不同的。这两种方法可靠地产生同样的败血症的结果。然而，通过，并通过技术的一个缺点是，它不便于控制的粪便量，盲肠被​​挤压时挤压。 </li><li>盲肠，然后轻轻挤压，挤出穿孔网站（6）少量的粪便。盲肠是返回到腹腔和腹膜6.0丝线缝合关闭。 （7）。 </li><li>皮肤反射7毫米剪辑（RS - 9258，Roboz手术器械）或米歇尔伤口剪辑（7毫米，RS - 9270）（8）关闭。 </li><li>复苏小鼠注射皮下注射1毫升预热0.9％生理盐水，使用25G针的解决方案。这种液体复苏措施，将引起败血症的高动力的阶段。这关键的一步是在讨论“部分进一步描述。 </li><li>可选：皮下注入丁丙诺啡（为0.05mg /公斤体重）或手术后镇痛的曲马多（20mg/kg体重）。被告知，这些阿片类药物的可以抑制呼吸和运动，这些影响可以被曲解为败血症的迹象。 </li><li>动物被暂时放在加热垫，或接触到的150W的红外线取暖灯或者立即返回到一个笼子里，直到他们从麻醉中恢复过来。恢复时间从30分钟至1小时。 </li><li>免费提供食物和水（水凝胶），放在笼底。 </li><li>小鼠进行监测，每12小时为一到两周的生存，或在不同的时间点不同的参数分析安乐死。 </li></ol>深水动物实验设计控制，将遵循不结扎和穿刺剖腹探查技术。 6至12小时后，手术过程中小鼠将成为昏昏欲睡和发展的发烧，立毛，腹泻，huddling，和不适，所有的败血症症状。非常严重的败血症小鼠可以勉强移动死前表现出体温的急剧下降。在这一步小鼠应安乐死，避免长时间的痛苦和磨难。 <img src="/files/ftp_upload/2860/2860fig0.jpg" alt="图0" /> 2。最常见的分析参数为了评估程序的结果，不同的参数，可以分析在器官，细胞提取物或体液。样品可以在不同时间点收集从3个小时的外科手术后一周。 <ol><li>小鼠的存活率。 </li><li>血清，腹腔及器官提取的细胞因子和趋化因子。 <ol><li> 白细胞介素6：生产和单核细胞，树突状细胞，巨噬细胞，B细胞，T细胞，粒细胞，肥大细胞和多细胞类型释放。在急性期反应和炎症反应中起着重要作用。 </li><li> 肿瘤坏死因子-α：Pleiotrophic起着核心作用，在炎症和细胞凋亡的细胞因子，。公司生产的单核细胞，巨噬细胞，中性粒细胞，树突状细胞和成纤维细胞。 </li><li> nterleukin -1β：由单核细胞，NK细胞，树突状细胞，B细胞和T细胞的产生。引起发热和急性期蛋白的合成。 </li><li> 白细胞介素10：促进吞噬细胞的摄取和Th2反应，但抑制antigEN介绍和Th1细胞促炎症反应。 </li><li> 白细胞介素10：促进吞噬细胞的摄取和Th2反应，但抑制抗原提呈和Th1细胞促炎症反应。 </li><li> 单核细胞趋化蛋白-1（也被称为CCL2）：新兵单核细胞，记忆性T细胞，树突状细胞和组织损伤和感染的网站。 </li><li> KC（CXCL1）：巨噬细胞和血管内皮细胞产生。鼠标KC是一个强有力的中性粒细胞的诱食和激活。 </li><li> RANTES的：一个单核细胞趋化。它可以chemoattract未刺激的CD4 + / CD45RO +记忆性T细胞和刺激的CD4 +和CD8 + T细胞与NA VE和记忆体的表型。 </li><li> 干扰素γ：由Th1细胞，细胞毒性T细胞，树突状细胞和自然杀伤细胞（NK）分泌的。提高在巨噬细胞抗原提呈和催化活性，并抑制Th2细胞的活性。促进白细胞迁移所需的附着力和约束力，促进NK细胞活性。 </li><li> 高流动性B组- 1蛋白在单核细胞是核因子κB激活提前糖化终产物（RAGE）的细胞受体结合，激活在败血症的后期阶段释放促炎介质。 </li></ol></li><li>作为衡量中性粒细胞浸润的髓过氧化物酶测定机关。髓过氧化物酶（MPO）是一个最丰富的中性粒细胞粒目前的过氧化物酶。这是一个存储azurophilic颗粒的中性粒细胞的溶酶体蛋白。 MPO有血红素色素，从而导致富含中性粒细胞的分泌物，脓和粘液的一些形式，如绿色。 </li><li>细菌器官和血液流加载，作为衡量每毫升菌落形成单位的数量。 </li></ol> 3。代表性的成果电过程最初是在C57BL / 6小鼠品系。我们测试的几个参数来调节通过改变结扎的长度和厚度的针， 如在图1和2所示的严重性败血症。在这两个因素中，结扎的长度似乎比针的厚度，以改变％的生存更有效。 如图1所示，增加结扎的长度超过1cm挑起与小鼠≤1厘米结扎相比，增加100％的死亡率。增加针的厚度也下降，从100％（使用22G针）的％生存到55％（使用19G针）与两个穿孔。我们还测试了在C57BL / 6和129SvJ小鼠中电的影响，以确定是否显示不同的小鼠品系相似或不同的CLP诱导的脓毒症的易感性。图3显示，在相同条件下，129SvJ老鼠比C57BL / 6表示％，死亡率增加，更容易感染。 <img src="/files/ftp_upload/2860/2860fig1.jpg" alt="图1" /> 图1。动物的生存盲肠结扎长度的影响 。电在C57BL / 6小鼠盲肠结扎使用两个不同长度的。这种单一的参数显着影响，因为与结扎的面积在不到3天死亡1厘米以上的所有组中的动物时，一组动物有一个约1厘米的结扎面积相比，动物的生存。 （N = 8）。 <img src="/files/ftp_upload/2860/2860fig2.jpg" alt="图2" /> 图2。针上生存的厚度的影响。电C57BL / 6小鼠使用两个大小不同的针，19G和22G。与盲肠穿孔使用19G针的小鼠表现出55-60％的生存。与此相反，小鼠盲肠穿孔使用22G针显示100％的生存，虽然他们经历了在第3-4天，炎症的典型症状消失后4天（N = 8）。 <img src="/files/ftp_upload/2860/2860fig3.jpg" alt="图3" /> 图3。比较脓毒症易感性之间的两个不同的小鼠品系 。CLP模型129SvJ和C57BL / 6相同的条件下进行。在评估期结束后129SvJ应变％的存活率为25％，比在C57BL / 6株少，说明一个更高的易感性多种微生物感染引起的炎症。 （N = 6）。

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没有利益冲突的声明。<br /&gt;动物实验，根据实验动物护理和使用指南“国家卫生研究所，动物护理和使用坦普尔大学委员会批准。

在这里，我们详细显示如何执行电在小鼠模型，并调节强度等级。 相比其他动物模型的败血症，中电可以执行在任何不同的年龄和性别的小鼠品系。它是一个相对容易和便宜的外科手术。在这种模式下，强度等级有直接影响生存的百分比。结扎的长度，厚度的穿刺针和数量等参数，可以控制调节的严重性/电死亡率。因此，获得戏剧性的变化中电严重性，用户可以改变结扎的?...

<table border="1"><tbody><tr><th>试剂名称</th><th>公司</th><th>目录编号</th><th>评论</th></tr><tr><td>氯胺酮</td><td> Ketaset </td><td> 0856-2013-01 </td><td></td></tr><tr><td>甲苯噻嗪</td><td> AnaSed </td><td> NADA＃139-236 </td><td></td></tr><tr><td>胰岛素注射器29G </td><td>埃克塞尔</td><td> 26028 </td><td></td></tr><tr><td>普理灵6-0丝线缝合， </td><td>爱惜康</td><td> 8680G </td><td></td></tr><tr><td> 19G和25G针</td><td>屋宇署</td><td> 305186 </td><td></td></tr><tr><td>剃须刀</td><td>一般供应</td><td>一般供应</td><td></td></tr><tr><td>红外线加热灯</td><td>一般供应</td><td>一般供应</td><td></td></tr><tr><td>米歇尔伤口剪辑7毫米</td><td> Roboz手术INSTR </td><td> RS - 9270 </td><td></td></tr><tr><td>耳挂面膜</td><td> Fisher Scientific则</td><td> 19-130-4181 </td><td></td></tr><tr><td>解剖剪刀</td><td> Roboz手术INSTR </td><td> RS - 6702 </td><td></td></tr><tr><td>优碘的解决方案</td><td> VWR </td><td> 63410-992 </td><td></td></tr><tr><td>手术钳</td><td> Roboz手术INSTR </td><td> RS - 5135 </td><td></td></tr><tr><td> 70％异丙醇垫</td><td> Fisher Scientific则</td><td> 22-031-350 </td><td></td></tr><tr><td>丁丙诺啡</td><td>贝德福德实验室</td><td> 55390-100-10 </td><td></td></tr><tr><td>曲马多</td><td> Sigma - Aldrich公司</td><td> 42965 </td><td></td></tr></tbody></table>

cecal ligation puncture procedure

人力败血症的特点是密集和大规模的感染，没有在本地主机中设置的全身反应。目前，在十大死亡原因，在美国重症监护病房 1中，败血症的行列。在败血症，有可能重叠的两个既定的血流动力学阶段。初始阶段（高动力）被定义为大规模生产的炎性细胞因子和活性氧的巨噬细胞和中性粒细胞，影响血管通透性导致低血压，心功能，导致代谢的变化，最终导致组织坏死和器官功能衰竭。因此，最常见的原因的死亡率是急性肾损伤。第二阶段（hypodynamic）是一个消炎的过程，涉及改变单核细胞抗原提呈，减少了淋巴细胞增殖功能和细胞凋亡增加。这种状态称为免疫抑制或免疫抑制大幅增加nocosomial感染和最终死亡的风险。这些病理生理过程的机制没有得到很好的特点。因为败血症的两个阶段，可能会导致不可逆的和不可弥补的损害，它是必不可少的，以确定患者的免疫和生理状态。这是许多治疗药物失败的主要原因。同一药物在不同的败血症阶段，可能是治疗或其他有害或有没有效果 2,3。了解在各级败血症关键是有一个合适的和全面的动物模型，再现了疾病的临床过程。这一点很重要的特点败血症过程中发生的病理生理机制，控制潜在的治疗药物测试模型条件。 为了研究人类败血症研究者的病因有不同的动物模型。临床使用最广泛的的模型是盲肠结扎穿孔（CLP）。电模式，使粪便材料释放进入腹膜腔，产生一个加剧的多种微生物感染引起的免疫反应的盲肠穿孔。这种模式符合人类生存条件，是临床相关。在人类，小鼠进行液体复苏的中华第一（初）高动力的阶段，在时间的推移到第二次（晚）hypodynamic阶段。此外，细胞因子的配置文件是类似人类败血症看到那里是增加淋巴细胞凋亡（4,5检讨） 。由于涉及多个重叠的败血症机制，研究人员需要一个合适的脓毒症模型控制的严重性，以取得一致的和可重复性的结果。

Watch this Scientific Journal Video about Cecal Ligation Puncture Procedure at JoVE.com

Cecal Ligation Puncture Procedure

作为一个有价值的工具，为研究人类败血症盲肠结扎穿刺的小鼠模型。

盲肠结扎穿刺过程

Human sepsis is characterized by a set of systemic reactions in response to intensive and massive infection that failed to be locally contained by the ...

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54.3K 次观看.  Temple University. 作为一个有价值的工具，为研究人类败血症盲肠结扎穿刺的小鼠模型。

视频: Cecal Ligation Puncture Procedure

Coronary Artery Ligation and Intramyocardial Injection in a Murine Model of Infarction

Mouse models are a valuable tool for studying acute injury and chronic remodeling of the myocardium in vivo. With the advent of genetic modifications to the whole organism or the myocardium and an array of biological and/or synthetic materials, there is great potential for any combination of these to assuage the extent of acute ischemic injury and impede the onset of heart failure pursuant to myocardial remodeling. 
Here we present the methods and materials used to reliably perform this microsurgery and the modifications involved for temporary (with reperfusion) or permanent coronary artery occlusion studies as well as intramyocardial injections. The effects on the heart that can be seen during the procedure and at the termination of the experiment in addition to histological evaluation will verify efficacy. 
Briefly, surgical preparation involves anesthetizing the mice, removing the fur on the chest, and then disinfecting the surgical area. Intratracheal intubation is achieved by transesophageal illumination using a fiber optic light. The tubing is then connected to a ventilator. An incision made on the chest exposes the pectoral muscles which will be cut to view the ribs. For ischemia/reperfusion studies, a 1 cm piece of PE tubing placed over the heart is used to tie the ligature to so that occlusion/reperfusion can be customized. For intramyocardial injections, a Hamilton syringe with sterile 30gauge beveled needle is used. When the myocardial manipulations are complete, the rib cage, the pectoral muscles, and the skin are closed sequentially. Line block analgesia is effected by 0.25% marcaine in sterile saline which is applied to muscle layer prior to closure of the skin. The mice are given a subcutaneous injection of saline and placed in a warming chamber until they are sternally recumbent. They are then returned to the vivarium and housed under standard conditions until the time of tissue collection. At the time of sacrifice, the mice are anesthetized, the heart is arrested in diastole with KCl or BDM, rinsed with saline, and immersed in fixative. Subsequently, routine procedures for processing, embedding, sectioning, and histological staining are performed. 
Nonsurgical intubation of a mouse and the microsurgical manipulations described make this a technically challenging model to learn and achieve reproducibility. These procedures, combined with the difficulty in performing consistent manipulations of the ligature for timed occlusion(s) and reperfusion or intramyocardial injections, can also affect the survival rate so optimization and consistency are critical.

Numerous genetic manipulations and/or intramyocardial injections of genes, proteins, cells, and/or biomaterials are superimposed upon the dimension of time in studies of acute ischemia/ reperfusion injury and chronic remodeling in mice. This video illustrates the microsurgical procedures for ischemia/reperfusion, permanent coronary artery ligation, and intramyocardial injection studies.

结扎冠状动脉内注射在小鼠模型梗死

Mouse models are a valuable tool for studying acute injury and chronic remodeling of the myocardium in vivo.  With the advent of genetic modifications to ...

科研

医学

Modified Technique for Coronary Artery Ligation in Mice

Myocardial infarction (MI) is one of the most important causes of mortality in humans1-3. In order to improve morbidity and mortality in patients with MI we need better knowledge about pathophysiology of myocardial ischemia. This knowledge may be valuable to define new therapeutic targets for innovative cardiovascular therapies4. Experimental MI model in mice is an increasingly popular small-animal model in preclinical research in which MI is induced by means of permanent or temporary ligation of left coronary artery (LCA)5. In this video, we describe the step-by-step method of how to induce experimental MI in mice.
The animal is first anesthetized with 2% isoflurane. The unconscious mouse is then intubated and connected to a ventilator for artificial ventilation. The left chest is shaved and 1.5 cm incision along mid-axillary line is made in the skin. The left pectoralis major muscle is bluntly dissociated until the ribs are exposed. The muscle layers are pulled aside and fixed with an eyelid-retractor. After these preparations, left thoracotomy is performed between the third and fourth ribs in order to visualize the anterior surface of the heart and left lung. The proximal segment of LCA artery is then ligated with a 7-0 ethilon suture which typically induces an infarct size ~40% of left ventricle. At the end, the chest is closed and the animals receive postoperative analgesia (Temgesic, 0.3 mg/50 ml, ip). The animals are kept in a warm cage until spontaneous recovery.

The surgical procedure used to induce experimental myocardial infarction in mice begins with left thoracotomy between the third and the fourth ribs in order to visualize the anterior surface of the heart and left lung. The left coronary artery is ligated, the chest is closed and the mouse is allowed to recover spontaneously.

改性技术在小鼠冠状动脉结扎

Myocardial infarction (MI) is one of the most important causes of mortality in humans1-3. In order to improve morbidity and mortality in patients with MI ...

Bile Duct Ligation in Mice: Induction of Inflammatory Liver Injury and Fibrosis by Obstructive Cholestasis

In most vertebrates, the liver produces bile that is necessary to emulsify absorbed fats and enable the digestion of lipids in the small intestine as well as to excrete bilirubin and other metabolic products. In the liver, the experimental obstruction of the extrahepatic biliary system initiates a complex cascade of pathological events that leads to cholestasis and inflammation resulting in a strong fibrotic reaction originating from the periportal fields. Therefore, surgical ligation of the common bile duct has become the most commonly used model to induce obstructive cholestatic injury in rodents and to study the molecular and cellular events that underlie these pathophysiological mechanisms induced by inappropriate bile flow. In recent years, different surgical techniques have been described that either allow reconnection or reanastomosis after bile duct ligation (BDL), e.g., partial BDL, or other microsurgical methods for specific research questions. However, the most frequently used model is the complete obstruction of the common bile duct that induces a strong fibrotic response after 21 to 28 days. The mortality rate can be high due to infectious complications or technical inaccuracies. Here we provide a detailed surgical procedure for the BDL model in mice that induce a highly reproducible fibrotic response in accordance to the 3R rule for animal welfare postulated by Russel and Burch in 1959.

Disruption of bile flow results in severe inflammatory cholestatic liver injury with a characteristic time-dependent sequence of morphological alterations. Here we present a protocol for the surgical ligation of the common bile duct in mice that allows to induce a strong fibrotic response after 21 to 28 days.

胆管结扎小鼠：感应炎性肝损伤和纤维化的阻塞性胆汁淤积

In most vertebrates, the liver produces bile that is necessary to emulsify absorbed fats and enable the digestion of lipids in the small intestine as well ...

Murine Left Anterior Descending (LAD) Coronary Artery Ligation: An Improved and Simplified Model for Myocardial Infarction

Ischemic heart disease (IHD), or acute coronary syndrome (ACS), is one of the leading causes of death in the United States. IHD is characterized by reduced blood supply to the heart, resulting in the loss of oxygen to and the ensuing necrosis of the heart muscle. The MI model has gained popularity for its use as a short-term ischemia-reperfusion model and a long-term permanent ligation model. Below, we describe a reliable method for the permanent ligation of the LAD. With mouse genetic engineering technology becoming more advanced, and with an increasing availability of quality murine surgical instruments, the mouse has become a popular model for MI surgeries. Our surgical model incorporates the use of an easily reversible anesthetic for the rapid recovery of the mouse; a minimally invasive endotracheal intubation without involving a tracheotomy; and a thoracentesis through the original thoracotomy site without creating an additional incision in the chest, as is done in some other methods, to effectively remove excess blood and air from the chest cavity. This method is comparatively less invasive than other methods, which dramatically reduces surgical and post-surgical complications and mortality and improves reproducibility.

Murine Left Anterior Descending LAD Coronary Artery Ligation: An Improved and Simplified Model for Myocardial Infarction

We provide a reliable method for left anterior descending artery (LAD) ligation in a mouse model. This method is comparatively less invasive than other methods, involving endotracheal intubation, a left-sided thoracotomy approach, and thoracentesis. This method can be used as a model for both acute and chronic myocardial infarction (MI).

鼠左前降支（LAD）冠状动脉结扎：改进和简化模型的心肌梗死

Ischemic heart disease (IHD), or acute coronary syndrome (ACS), is one of the leading causes of death in the United States. IHD is characterized by ...

Rat Model of the Associating Liver Partition and Portal Vein Ligation for Staged Hepatectomy (ALPPS) Procedure

Recent clinical data support an aggressive surgical approach to both primary and metastatic liver tumors. For some indications, like colorectal liver metastases, the amount of liver tissue left behind after liver resection has become the main limiting factor of resectability of large or multiple liver tumors. A minimal amount of functional tissue is required to avoid the severe complication of post-hepatectomy liver failure, which has high morbidity and mortality. Inducing liver growth of the prospective remnant prior to resection has become more established in liver surgery, either in the form of portal vein embolization by interventional radiologists or in the form of portal vein ligation several weeks prior to resection. Recently, it was shown that liver regeneration is more extensive and rapid, when the parenchymal transection is added to portal vein ligation in a first stage and then, after only one week of waiting, resection performed in a second stage (Associating Liver Partition and Portal vein ligation for Staged hepatectomy = ALPPS). ALPPS has rapidly become popular across the world, but has been criticized for its high perioperative mortality. The mechanism of accelerated and extensive growth induced by this procedure has not been well understood. Animal models have been developed to explore both the physiological and molecular mechanisms of accelerated liver regeneration in ALPPS. This protocol presents a rat model that allows mechanistic exploration of accelerated regeneration.

Rat Model of the Associating Liver Partition and Portal Vein Ligation for Staged Hepatectomy ALPPS Procedure

Inducing rapid liver hypertrophy using Associating Liver Partition and Portal vein ligation for a Staged hepatectomy (ALPPS) has been proposed for resection of borderline resectable liver tumors. This model may elucidate mechanisms involved in rapid hypertrophy and allows testing of drugs that promote or block the acceleration of regeneration.

肝分型与门静脉结扎大鼠分期肝切除 (ALPPS) 手术模型的建立

Recent clinical data support an aggressive surgical approach to both primary and metastatic liver tumors. For some indications, like colorectal liver ...

Puncture-Induced Iris Neovascularization as a Mouse Model of Rubeosis Iridis

We describe a model of puncture-induced iris neovascularization as a general model for noninvasive evaluation of angiogenesis. The model is also relevant for targeting neovascular glaucoma, a sight-threatening complication of diabetic retinopathy. This method is based on the induction of iris vascular response by a series of self-sealing uveal punctures on BALB/c mice and takes advantage of the postpartum maturation of mouse ocular vasculature. Mouse pups undergo uveal punctures from postnatal day 12.5, when the pups naturally open their eyes, until postnatal day 24.5. Due to the transparency of the cornea, iris vasculature can be analyzed easily through time by noninvasive in vivo methods. Furthermore, the semitransparent iris of BALB/c mice can be flatmounted for detailed immunohistologic analysis with minimal non-specific background staining. In this model, angiogenesis is mainly driven by the inflammatory and plasminogen activating systems. The puncture-induced model is the first to induce iris neovascularization in small rodents, and has the advantage of allowing direct noninvasive in vivo analysis of the angiogenic process. Moreover, the model can be combined with angiogenic modulating substances, which highlights its potential in the study of angiogenesis with an in vivo perspective.

Iris neovascularization, a common complication of ischemic retinal disease, may lead to sight-threatening neovascular glaucoma. Here, we describe a murine protocol for inducing experimental iris neovascularization that may be used for noninvasive evaluation of angiogenesis-modulating substances.

穿刺诱导的虹膜新生血管虹膜虹膜小鼠模型的建立

We describe a model of puncture-induced iris neovascularization as a general model for noninvasive evaluation of angiogenesis. The model is also relevant ...

Subclavian Vein Puncture As an Alternative Method of Blood Sample Collection in Rats

Blood collection with enough blood volume is essential in animal experiments. Blood collection from the tail vein of rats is popular and less stressful compared to other more aggressive methods such as retro-orbital plexus sample collection. However, this blood collection method is sometimes limited by an unsatisfactory success rate. Here, we introduce a method for blood collection through the subclavian vein puncture. The subclavian vein is located just under the clavicle and this vein is large enough to fulfill the volume requirements of blood collection. Our results show that this method is safe and applicable for blood collection sampling with the required blood volume. Blood collection through the subclavian vein puncture could serve as an alternative blood collection method in case of failed tail vein blood sampling in rats.

Here, we present a protocol to collect blood samples from the rat subclavian vein.

锁骨下静脉穿刺作为大鼠采血的一种替代方法

Blood collection with enough blood volume is essential in animal experiments. Blood collection from the tail vein of rats is popular and less stressful ...

Blood Collection Through Subclavian Vein Puncture in Mice

The mouse is the foremost mammalian model for studying human disease and human health. However, blood sample collection from mice is challenging in research work. Tail blood collection is a popular method when a small amount of blood sample is needed. Orbital artery could be considered if a large amount of blood is needed but this blood collection method has ethical issues. Formerly, we demonstrated the feasibility and safety of blood sample collection through subclavian vein puncture in rats, and here we investigate whether this method could be used in mice. We report that this method is safe and practical for blood collection in mice. Blood collection through the subclavian vein puncture in mice can be a convenient method in daily research works.

Here, we present a protocol to take blood samples from the subclavian vein of mice.

小鼠锁骨下静脉穿刺采血

The mouse is the foremost mammalian model for studying human disease and human health. However, blood sample collection from mice is challenging in ...

Left Coronary Artery Ligation: A Surgical Murine Model of Myocardial Infarction

Ischemic heart disease and subsequent myocardial infarction (MI) is one of the leading causes of mortality in the United States and around the world. In order to explore the pathophysiological changes after myocardial infarction and design future treatments, research models of MI are required. Permanent ligation of the left coronary artery (LCA) in mice is a popular model to investigate cardiac function and ventricular remodeling post MI. Here we describe a less invasive, reliable, and reproducible surgical murine MI model by permanent ligation of the LCA. Our surgical model comprises of an easily reversible general anesthesia, endotracheal intubation that does not require a tracheotomy, and a thoracotomy. Electrocardiography and troponin measurement should be performed to ensure MI. Echocardiography at day 28 after MI will discern heart function and heart failure parameters. The degree of cardiac fibrosis can be evaluated by Masson's trichrome staining and cardiac MRI. This MI model is useful for studying the pathophysiological and immunological alterations after MI.

Presented here is a surgical procedure for permanent ligation of the left coronary artery in mice. This model can be used to investigate the pathophysiology and associated inflammatory response after myocardial infarction.

左冠状动脉结扎术：心肌梗死的外科小鼠模型

Ischemic heart disease and subsequent myocardial infarction (MI) is one of the leading causes of mortality in the United States and around the world. In ...

Tail Vein Puncture Without Anesthesia

To begin, secure the rat in a plastic restraining cone, enclose the large end of the cone around the base of the tail. Ensure that the animal is comfortable and breathing is unrestricted throughout the procedure. Dip the tail in warm water for about one minute to dilate the vein, and dry the tail with a paper towel.In the restrainer, position the animal facing down with the tail lying on a heating pad. To select the appropriate tail vein for sampling, rotate the entire animal to the right or left side. Use the terminal third of the tail for blood vessel puncturing since the vessels become more superficial in this zone.Wipe the tail with 70%ethanol wipes at the puncture site and place it on the edge of the heating pad, creating an angle in the terminal third of the tail. This brings the vein to the surface and creates more space for taking the sample. For blood sampling, move the plunger back and forth in the syringe several times to smoothen the withdrawal and then pull on the plunger to create negative pressure in the syringe.Secure the tail flat on the heating pad using the non-dominant index and middle finger. Place the middle finger proximally and the index finger distally with the puncture site between these two fingers. Slide the insulin syringe against the index finger with the eye of the needle pointing upward until it is inserted into the vein.This creates an angle of 35 degrees between the needle and the tail. Once the needle has entered the vein, blood will flow into the syringe. Slowly withdraw the plunger of the syringe at a steady rate to collect the desired volume.Remove the syringe from the tail and allow a blood drop to form on the puncture side of the tail. Aspirate this blood to collect a few more microliters of blood, in case of a non-sterile procedure. Apply pressure to the puncture site to stop the bleeding and wipe the area with a new alcohol wipe.After removing the rat from the cone, return it to its cage. The success rates with the tail vein puncture methods were compared with the success rate of the penile vein puncture method under anesthesia. Interestingly, under anesthesia, the tail vein became very unreliable and had a success rate of only 25%The success rates showed almost similar results for the tail vein puncture in conscious rats and the penile vein puncture under anesthesia.The blood sampling was performed in less than six minutes in the tail vein puncture without anesthesia, and the penile vein puncture method under anesthesia, whereas the tail vein puncture method under anesthesia took more than eight minutes due to multiple failures.