我们感谢西尔维·为该协议的的关键的读数的勒萨热和的所有劳顾会委员。这项工作被受支持的由一个从的作者:卫生研究所（CIHR）的的加拿大研究院的的授出日期（澳门币-77545）。纳萨莉拉布雷克被由一个FRSQ的高级奖学金的的支持，，和MELISSA的Mathieu收到1 NSERC亚历山大·格雷厄姆·加拿大贝尔公司研究生奖学金。

1。手术前的准备工作根据加拿大动物护理准则理事会处理小鼠。 <ol><li> anesthetise小鼠注射氯胺酮/甲苯​​噻嗪（150/300毫克/千克，IP）或异​​氟醚吸入（2％，1L氧气）。如果可以，异氟醚应该使用，因为它允许为更好地控制麻醉时间和深度。此外，氧管理的同时，支持动物的生理功能。此外，异氟醚直接消除肺部，因此并不需要肝，肾代谢。 </li><li>注入丁丙诺啡（0.05 - 0.1毫克/千克，SC）或另一种止​​痛药。 </li><li>适用于鼠标的眼睛上的药膏，手术前，以避免眼睛干涩（如果使用异氟醚）。 </li><li>确认鼠标是睡着了捏它的爪子，滚它的尾巴。如果鼠标没有反应，进行手术。 </li></ol> 2。手术<p类="jove_content"&gt;很可能浅表淋巴结（LN），通过手术移除4:2腹股沟和2臂丛。 LN的隔离更容易在年轻瘦老鼠，因为在老年小鼠的脂肪组织的存在（6-8周）可与LN隔离干扰。然而，这一限制被克服，反复练习。为的小鼠舒适，我们收获一次最多2淋巴结，当我们执行时间课程实验，我们收获LNS每隔一天。此外，为防止感染，抗生素可以给鼠标，但我们不这样做，伤口愈合得很好。一个需要注意，根据实验设计，使用抗生素，可能会影响观察的结果（例如，如果细菌的反应测量）。请注意，所有仪器每次使用后消毒和无菌条件下保持整个程序。 <ol><li>鼠标放在其副作用。 </li><li>本地化的地区，在那里你会执行incisioN，这取决于你想收获的法律公告。 （ 图1）。 </li><li>洗必泰，或另一种消毒剂，适用于该地区。 </li><li>请用锋利的剪刀（约5毫米），无论是在前肢背部或后肢腹部附近（ 图1）以上的微小切口。 </li><li>删除松散的头发，用钳子切断。如果你愿意，你可以刮胡子鼠标，然后才作出的切口。但是，一个较长的过程，我们发现，这是没有必要以来，如下所示，它不会改善愈合，减少感染，这都是非常罕见的。 </li><li> 2钳拉伸切口才能看到的LN。切口可能达到10毫米。号法律公告出现灰褐色或暗比周围脂肪。 </li><li>捏用一个镊子的LN上的筋膜（薄膜覆盖的脂肪和组织），轻轻拉，不破坏周围组织。 </li><li>将第二个钳尽可能underneath的LN。第一镊子，打破筋膜和删除号法律公告。如果您已成功收获的LN，染色的血液应该出现在其中的LN以前所在的位置。需要注意的是等渗溶液中的LN汇。这个简单的测试，让您验证您已提取的一种法律公告，而不是脂肪组织。 </li><li>确认没有毛发在伤口。 </li><li>坚持2部分皮肤（皮肤内），装订剪辑，关闭切口。我们使用应用镊子一些米​​歇尔夹。安装1或2个片段，切口的大小而定。大多数情况下，剪辑时脱落的组织愈合。 </li></ol> 3。术后护理<ol><li>笼底部添加一些湿的食物，手术后容易使老鼠能养活。 </li><li>手术后约6至8小时，检查，以确保他们没有经历过痛苦的小鼠。仔细观察他们的行为（姿势，步态，毛皮的外观与同类和相互作用）。如果小鼠的整体外观差，管理的丁丙诺啡第二剂量。由老鼠第二天应该恢复其正常的行为，否则，继续丁丙诺啡的管理。小鼠从未达到先前确定的，由动物保健理事会接受的终点。你可能想寻求咨询和帮助你照顾动物饲养箱技术员。 </li></ol> 4。淋巴结处理<ol><li>在实验室中，获得单细胞悬液，游离为您常​​用的程序的法律公告。我们LN和对磨砂幻灯片按常规获得1-2×10 6细胞每号法律公告。另外，我们准备RNA提取的LN。 </li><li>细胞染色为你平时的协议。你应该获得预期的LN流式细胞仪配置文件。 </li></ol> 5。代表结果例如前部和侧散射带够很多经手术或从牺牲的鼠标（ 图2）从麻醉鼠标收获淋巴结。在现场淋巴细胞门的形象和细胞的百分比相同，表明法律公告手术是一个合适的技术，收获细胞。 <img alt="图1" src="/files/ftp_upload/3444/3444fig1.jpg" /> 图1。腹股沟和鼠标臂LN的本地化。 A.腹股沟号法律公告位于略高于后肢，稍往肚子。 B.臂丛LN是位于前腿后面。 <img alt="图2" src="/files/ftp_upload/3444/3444fig2.jpg" /> LN的手术后得到图2。流量淋巴细胞流式细胞仪轮廓。前部和侧散射剖面显示从麻醉收获的淋巴结（一）经手术或从牺牲的鼠标（二）etized鼠标。淋巴结收获，分离和流式细胞仪分析。细胞在活的淋巴细胞门的百分比显示。

<ol>
	<li>Zehn, D., Lee, S. Y., Bevan, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Complete+but+curtailed+T-cell+response+to+very+low-affinity+antigen.">Complete but curtailed T-cell response to very low-affinity antigen.</a> Nature. 458, 211-214 (2009).</li><li>Vezys, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Memory+CD8+T-cell+compartment+grows+in+size+with+immunological+experience.">Memory CD8 T-cell compartment grows in size with immunological experience.</a> Nature. 457, 196-199 (2009).</li><li>Zhou, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differentiation+and+persistence+of+memory+CD8(+)+T+cells+depend+on+T+cell+factor+1.">Differentiation and persistence of memory CD8(+) T cells depend on T cell factor 1.</a> Immunity. 33, 229-240 (2010).</li><li>Wirth, T. 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Immunol. 182, 6195-6206 (2009).</li><li>Rooke, R., Waltzinger, C., Benoist, C., Mathis, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Targeted+complementation+of+MHC+class+II+deficiency+by+intrathymic+delivery+of+recombinant+adenoviruses.">Targeted complementation of MHC class II deficiency by intrathymic delivery of recombinant adenoviruses.</a> Immunity. 7, 123-134 (1997).</li><li>Witherden, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tetracycline-controllable+selection+of+CD4(+)+T+cells:+half-life+and+survival+signals+in+the+absence+of+major+histocompatibility+complex+class+II+molecules.">Tetracycline-controllable selection of CD4(+) T cells: half-life and survival signals in the absence of major histocompatibility complex class II molecules.</a> J. Exp. 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没有利益冲突的声明。

我们已经描述了LN的手术协议，它可以适用于许多实验系统。虽然非常有益的免疫反应或免疫细胞的平衡，这种技术具有一些限制。首先，只有四个LNS可以收获。二，免疫反应的研究必须是系统性的，或发生在腹股沟浅淋巴结或臂丛（皮肤排出）。最后，收获细胞数号法律公告搬迁的质量，技术的限制，这可能有助于实验变异时需要绝对细胞数的影响。然而，对靶细胞的百分比，对自己，可以提供相关和准确的信息，特别是当LN的大小并不各种实验条件各不相同。幸运的是，一旦技术以及掌握，LN的去除率提高质量，从而在每个LN的高度重复性的手机号码。我们相信，尽管这几年的局限性，法律公告手术提供了重要的优势，研究免疫反应的进展。首先，随着时间的推移的免疫反应，可以在一个特定的鼠标跟踪，减少实验的变异性以及使用的老鼠数量。二，如淋巴细胞稳态或免疫反应开始发生在LNS的事件，可直接影响。另一个主要优点是可以从比从延长细胞的特性，允许血液淋巴结恢复，更多的细胞。该技术也适用于其他用途，如转基因小鼠的基因分型或研究通过淋巴结，如B细胞，树突状细胞和巨噬细胞的其他细胞群体居住或贩运。 我们的实验室和其他已使用了好几年这项技术，我们从来没有观察7-9,12,13的免疫反应过程中的任何干扰免疫稳态5-7。休息野生型C57BL / 6小鼠的序列号法律公告手术没有影响的比例和数量的CD4 + </s5-7起来&gt;和CD8 + T细胞。此外，它也不会影响这些参数中残留的淋巴结。此外，在免疫反应过程中，浅表性LN的拆除并不影响T细胞的反应，也不产生记忆性T细胞8,9,12,13动力学。此外，方方T细胞的反应比较之后要么顺序号法律公告手术或处死小鼠没有表现出任何差异（未发表的结果和图2）在我们的手中。因此，序列号法律公告手术是合适的研究T细胞的动态平衡和响应。

<table border="1"><tbody><tr><td> 试剂名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论 </td></tr><tr><td>格雷夫精细模式高级钳</td><td>哈佛仪器</td><td> 52-2144 </td><td>强烈弯曲，10厘米（4英寸），0.8毫米尖</td></tr><tr><td>米歇尔剪辑应用和删除钳</td><td>哈佛仪器</td><td> 52-3779 </td><td> 12.5厘米（5） </td></tr><tr><td>米歇尔剪辑100 </td><td>哈佛仪器</td><td> 52-3746 </td><td> 7.5-1.75毫米</td></tr><tr><td> temgesic（盐酸丁丙诺啡） </td><td>先灵葆雅</td><td></td><td></td></tr><tr><td> vetalar（盐酸氯胺酮） </td><td> boniche动物保健</td><td></td><td>根据DIN:01989529 </td></tr><tr><td> rompun（二甲苯胺噻嗪） </td><td>拜耳</td> &lt;TD&gt; </td><td>根据DIN:02169592 </td></tr><tr><td>异氟醚</td><td>雅培</td><td></td><td>根据DIN:02032384 </td></tr><tr><td> hypotears眼膏</td><td>诺华Ophtalmics </td><td></td><td>根据DIN:02133288 </td></tr><tr><td> baxedin（Clorhexidine葡萄糖2％，异丙醇70％） </td><td>欧米茄</td><td> L0000017 </td><td>根据DIN:02251477 </td></tr></tbody></table>

murine superficial lymph node surgery

在免疫学领域，了解进展对疫苗，感染或肿瘤的免疫反应，反应之后往往随着时间的推移。同样，淋巴细胞稳态的研究需要时间课程实验。这些研究是在相同的鼠标理想，以减少实验的变异，以及使用的老鼠数量。抽血时间课程实验允许的性能，但它只能提供约外周血淋巴细胞的信息，并提供数量有限的细胞1-4。由于通过人体循环和居住在淋巴结中的淋巴细胞有不同的特性，重要的是要检查这两个地方。淋巴结顺序切除手术提供了一个独特的机会，后续的免疫反应或免疫细胞膨胀，随着时间的推移相同的鼠标。此外，这种技术的产生，这是足够的PE 1-2X10 6个淋巴结细胞之间rform表型特征和/或功能分析。顺序淋巴结手术或淋巴结清扫已成功地应用于我们和其他国家5-11。在这里，我们描述如何可以通过一个小切口，在皮肤一个anesthetised鼠标删除臂丛和腹股沟淋巴结肿大。由于手术是肤浅的，并迅速完​​成，鼠标恢复速度非常快，愈合好，不会遇到过多的疼痛。每隔一天，有可能收获一个或两个淋巴结允许时间课程实验。因此，该技术是适合学习节点居住淋巴细胞淋巴结的特点，随着时间的推移。这种方法适用于各种实验设计，我们相信，许多实验室将受益于执行顺序淋巴结手术。

Watch this Scientific Journal Video about Murine Superficial Lymph Node Surgery at JoVE.com

Murine Superficial Lymph Node Surgery

遵循多时间内相同的鼠标免疫反应的进展，淋巴结可以依次通过手术切除。在这里，我们描述了这种技术可以如何执行。

小鼠浅表淋巴结手术

In the field of immunology, to understand the progression of an immune response against a vaccine, an infection or a tumour, the response is often ...

Nanyang Technological University

Research

JoVE Journal

Immunology and Infection

42.1K 次观看.  Maisonneuve-Rosemont Hospital Research Center. 遵循多时间内相同的鼠标免疫反应的进展，淋巴结可以依次通过手术切除。在这里，我们描述了这种技术可以如何执行。

视频: Murine Superficial Lymph Node Surgery

Intravital Microscopy of the Inguinal Lymph Node

Lymph nodes (LN's), located throughout the body, are an integral component of the immune system. They serve as a site for induction of adaptive immune response and therefore, the development of effector cells. As such, LNs are key to fighting invading pathogens and maintaining health. The choice of LN to study is dictated by accessibility and the desired model; the inguinal lymph node is well situated and easily supports studies of biologically relevant models of skin and genital mucosal infection.
The inguinal LN, like all LNs, has an extensive microvascular network supplying it with blood. In general, this microvascular network includes the main feed arteriole of the LN that subsequently branches and feeds high endothelial venules (HEVs). HEVs are specialized for facilitating the trafficking of immune cells into the LN during both homeostasis and infection. How HEVs regulate trafficking into the LN under both of these circumstances is an area of intense exploration. The LN feed arteriole, has direct upstream influence on the HEVs and is the main supply of nutrients and cell rich blood into the LN. Furthermore, changes in the feed arteriole are implicated in facilitating induction of adaptive immune response. The LN microvasculature has obvious importance in maintaining an optimal blood supply to the LN and regulating immune cell influx into the LN, which are crucial elements in proper LN function and subsequently immune response. 
The ability to study the LN microvasculature in vivo is key to elucidating how the immune system and the microvasculature interact and influence one another within the LN. Here, we present a method for in vivo imaging of the inguinal lymph node. We focus on imaging of the microvasculature of the LN, paying particular attention to methods that ensure the study of healthy vessels, the ability to maintain imaging of viable vessels over a number of hours, and quantification of vessel magnitude. Methods for perfusion of the microvasculature with vasoactive drugs as well as the potential to trace and quantify cellular traffic are also presented. 
Intravital microscopy of the inguinal LN allows direct evaluation of microvascular functionality and real-time interface of the direct interface between immune cells, the LN, and the microcirculation. This technique potential to be combined with many immunological techniques and fluorescent cell labelling as well as manipulated to study vasculature of other LNs.

A technique for performing intravital microscopy of the inguinal lymph node (LN) is outlined. Such technique allows for real-time, in vivo study of the lymph node microvasculature and structure both during homeostasis and infection. This technique can be adapted to cell trafficking studies and to other lymph node sites.

腹股沟淋巴结的活体显微镜

Lymph nodes (LN's), located throughout the body, are an integral component of the immune system. They serve as a site for induction of adaptive immune ...

科研

免疫与感染

Intralymphatic Immunotherapy and Vaccination in Mice

Vaccines are typically injected subcutaneously or intramuscularly for stimulation of immune responses. The success of this requires efficient drainage of vaccine to lymph nodes where antigen presenting cells can interact with lymphocytes for generation of the wanted immune responses. The strength and the type of immune responses induced also depend on the density or frequency of interactions as well as the microenvironment, especially the content of cytokines. As only a minute fraction of peripherally injected vaccines reaches the lymph nodes, vaccinations of mice and humans were performed by direct injection of vaccine into inguinal lymph nodes, i.e.&#160;intralymphatic injection. In man, the procedure is guided by ultrasound. In mice, a small (5-10 mm) incision is made in the inguinal region of anesthetized animals, the lymph node is localized and immobilized with forceps, and a volume of 10-20 &#956;l of the vaccine is injected under visual control. The incision is closed with a single stitch using surgical sutures. Mice were vaccinated with plasmid DNA, RNA, peptide, protein, particles, and bacteria as well as adjuvants, and strong improvement of immune responses against all type of vaccines was observed. The intralymphatic method of vaccination is especially appropriate in situations where conventional vaccination produces insufficient immunity or where the amount of available vaccine is limited.

Prophylactic and therapeutic vaccination often fails to stimulate strong immune responses due to week drainage of the vaccine to lymph nodes and consequently poor involvement of immune cells. By direct injection of vaccine to lymph nodes, so-called intralymphatic injection, vaccine efficacy can be strongly improved and vaccine doses can be reduced.

淋巴免疫治疗和疫苗接种小鼠

Vaccines are typically injected subcutaneously or intramuscularly for stimulation of immune responses. The success of this requires efficient drainage of ...

Surgical Technique for Superior Cervical Ganglionectomy in a Murine Model

Growing evidence suggests that the sympathetic nervous system plays an important role in cancer progression. Adrenergic innervation regulates salivary gland secretion, circadian rhythm, macular degeneration, immune function, and cardiac physiology. Murine surgical sympathectomy is a method for studying the effects of adrenergic innervation by allowing for complete, unilateral adrenergic ablation while avoiding the need for repeated pharmacologic intervention and the associated side effects. However, surgical sympathectomy in mice is technically challenging because of the small size of the superior cervical ganglion. This study describes a surgical technique for reliably identifying and resecting the superior cervical ganglion to ablate the sympathetic nervous system. The successful identification and removal of the ganglion are validated by imaging the fluorescent sympathetic ganglia using a transgenic mouse, identifying post-resection Horner's syndrome, staining for adrenergic markers in the resected ganglia, and observing diminished adrenergic immunofluorescence in the target organs following sympathectomy. This model enables future studies of cancer progression as well as other physiological processes regulated by the sympathetic nervous system.

The present protocol describes a mouse model of the ablation of adrenergic innervation by identifying and resecting the superior cervical ganglion.

小鼠模型中上颈神经节切除术的手术技术

Growing evidence suggests that the sympathetic nervous system plays an important role in cancer progression. Adrenergic innervation regulates salivary ...

癌症研究

Modified Laparoscopic Inguinal Lymphadenectomy Using an Abdominal Subcutaneous Approach

Inguinal lymph node metastases significantly impact the prognosis of patients with penile cancer. Therefore, timely inguinal lymph node dissection is essential for the comprehensive treatment of penile cancer. Compared with the traditional open inguinal lymphadenectomy, laparoscopic inguinal lymphadenectomy offers similar tumor control with fewer complications. The current techniques for the laparoscopic surgical approach include L-lymphoid clearance sequence and preservation of the great saphenous vein. In this study, a transabdominal subcutaneous anterograde approach was employed to improve laparoscopic inguinal lymph node dissection and conserve the great saphenous vein, resulting in favorable outcomes. Furthermore, only 2 out of 21 patients experienced wound infections, and only 1 exhibited lymphatic leakage from the drainage orifice. These findings indicate that the use of an innovative subcutaneous transperitoneal retrograde approach is safe for abdominal endoscopic common iliac plexus dissection with fewer complications in patients with penile cancer compared with traditional open surgery. Notably, the postoperative survival rate of penile cancer patients is significantly influenced by the presence or absence of inguinal lymph node metastasis and the extent of metastasis. Timely inguinal lymph node dissection is essential since it significantly impacts the treatment of penile cancer. Besides, laparoscopic inguinal lymphadenectomy offers comparable tumor control to open surgery with significantly reduced complications. Notably, standardized approaches for laparoscopic surgery, cleaning procedures, and preservation of the saphenous vein are crucial for inguinal lymph node dissection. The laparoscopic inguinal lymphadenectomy technique can be improved by employing the abdominal subcutaneous anterograde approach. This article provides a comprehensive account of the procedures and technical improvements associated with the modified laparoscopic inguinal lymphadenectomy using the abdominal subcutaneous approach.

The efficacy of laparoscopic inguinal lymph node dissection is comparable to that of open surgery. It also significantly decreases the incidence of complications. In this study, a modified laparoscopic method was used for inguinal lymph node dissection through the abdominal subcutaneous approach for penile cancer treatment.

使用腹部皮下入路的改良腹腔镜腹股沟淋巴结清扫术

Inguinal lymph node metastases significantly impact the prognosis of patients with penile cancer. Therefore, timely inguinal lymph node dissection is ...

Laparoscopic Technique for Serial Collection of Liver and Mesenteric Lymph Nodes in Macaques

The mesenteric lymph nodes (MLN) and the liver are exposed to microbes and microbial products from the gastrointestinal (GI) tract, making them immunologically unique. The GI tract and associated MLN are sites of early viral replication in human immunodeficiency virus (HIV) infection and the MLN are likely important reservoir sites that harbor latently-infected cells even after prolonged antiretroviral therapy (ART). The liver has been shown to play a significant role in immune responses to lentiviruses and appears to play a significant role in clearance of virus from circulation. Nonhuman primate (NHP) models for HIV and Acquired Immunodeficiency Syndrome (AIDS) closely mimic these aspects of HIV infection and serial longitudinal sampling of primary sites of viral replication and the associated immune responses in this model will help to elucidate critical events in infection, pathogenesis, and the impact of various intervention strategies on these events. Current published techniques to sample liver and MLN together involve major surgery and/or necropsy, which limits the ability to investigate these important sites in a serial fashion in the same animal. We have previously described a laparoscopic technique for collection of MLN. Here, we describe a minimally invasive laparoscopic technique for serial longitudinal sampling of liver and MLN through the same two port locations required for the collection of MLN. The use of the same two ports minimizes the impact to the animals as no additional incisions are required. This technique can be used with increased sampling frequency compared to major abdominal surgery and reduces the potential for surgical complications and associated local and systemic inflammatory responses that could complicate interpretation of results. This procedure has potential to facilitate studies involving NHP models while improving animal welfare.

Here, we describe a minimally invasive laparoscopic technique for serial sampling of liver and mesenteric lymph nodes (MLN) in macaques that allows for increased sampling frequency, and reduces the potential for surgical complications when compared to performing a laparotomy.

腹腔镜技术在猕猴肝和肠系膜淋巴结的串行采集

The mesenteric lymph nodes (MLN) and the liver are exposed to microbes and microbial products from the gastrointestinal (GI) tract, making them ...

医学

Blocking Lymph Flow by Suturing Afferent Lymphatic Vessels in Mice

Lymphatic vessels are critical in maintaining tissue fluid balance and optimizing immune protection by transporting antigens, cytokines, and cells to draining lymph nodes (LNs). Interruption of lymph flow is an important method when studying the function of lymphatic vessels. The afferent lymphatic vessels from the murine footpad to the popliteal lymph nodes (pLNs) are well-defined as the only routes for lymph drainage into the pLNs. Suturing these afferent lymphatic vessels can selectively prevent lymph flow to the pLNs. This method allows for interference in lymph flow with minimal damage to the lymphatic endothelial cells in the draining pLN, the afferent lymphatic vessels, as well as other lymphatic vessels around the area. This method has been used to study how lymph impacts high endothelial venules (HEV) and chemokine expression in the LN, and how lymph flows through the adipose tissue surrounding the LN in the absence of functional lymphatic vessels. With the growing recognition of the importance of lymphatic function, this method will have broader applications to further unravel the function of lymphatic vessels in regulating the LN microenvironment and immune responses.

A protocol to block lymph flow by surgical suturing of afferent lymphatic vessels is presented.

通过在小鼠中吸液性淋巴血管阻止淋巴流

Lymphatic vessels are critical in maintaining tissue fluid balance and optimizing immune protection by transporting antigens, cytokines, and cells to ...

A Revised Method for Inducing Secondary Lymphedema in the Hindlimb of Mice

Animal models are of paramount importance in the research of lymphedema in order to understand the pathophysiology of the disease but also to explore potential treatment options. This mouse model allows researchers to induce significant lymphedema lasting at least 8 weeks. Lymphedema is induced using a combination of fractioned radiotherapy and surgical ablation of lymphatics. This model requires that the mice get a dose of 10 Gray (Gy) radiation before and after surgery. The surgical part of the model involves ligation of three lymph vessels and extraction of two lymph nodes from the mouse hindlimb. Having access to microsurgical tools and a microscope is essential, due to the small anatomical structures of mice. The advantage of this model is that it results in statistically significant lymphedema, which provides a good basis for evaluating different treatment options. It is also a great and easily available option for microsurgical training. The limitation of this model is that the procedure can be time consuming, especially if not practiced in advance. The model results in objectively quantifiable lymphedema in mice, without causing severe morbidity and has been tested in three separate projects.

This animal model enables researchers to induce statistically significant secondary lymphedema in the hindlimb of mice, lasting at least 8 weeks. The model can be used to study the pathophysiology of lymphedema and to investigate novel treatment options.

小鼠下肢诱导继发性淋巴水肿的修订方法

Animal models are of paramount importance in the research of lymphedema in order to understand the pathophysiology of the disease but also to explore ...

评估淋巴结转移过程中抗原特异性 CD8+ T 细胞

Draining Lymph Node Metastasis Model for Assessing the Dynamics of Antigen-Specific CD8+ T Cells During Tumorigenesis

Tumor antigen-specific CD8+ T cells from draining lymph nodes gain an accumulating importance in mounting anti-tumor immune response during tumorigenesis. However, in many cases, cancer cells form metastatic loci in lymph nodes before further metastasizing to distant organs. To what extent the local and systematic CD8+ T cell responses were influenced by LN metastasis remains obscure. To this end, we set up a murine LN metastasis model combined with a B16F10-GP melanoma cell line expressing the surrogate neoantigen derived from lymphocytic choriomeningitis virus (LCMV), glycoprotein (GP), and P14 transgenic mice harboring T cell receptors (TCRs) specific to GP-derived peptide GP33-41 presented by the class I major histocompatibility complex (MHC) molecule H-2Db. This protocol enables the study of antigen-specific CD8+ T cell responses during LN metastasis. In this protocol, C57BL/6J mice were subcutaneously implanted with B16F10-GP cells, followed by adoptive transfer with naive P14 cells. When the subcutaneous tumor grew to approximately 5 mm in diameter, the primary tumor was excised, and B16F10-GP cells were directly injected into the tumor draining lymph node (TdLN). Then, the dynamics of CD8+ T cells were monitored during the process of LN metastasis. Collectively, this model has provided an approach to precisely investigate the antigen-specific CD8+ T cell immune responses during LN metastasis.

作者聚焦：研究 LN 转移期间 CD8+ T 细胞全身和局部动力学的模型

The experimental design presented here provides a useful reproductive model for the studies of antigen-specific CD8+ T cells during lymph node (LN) metastasis, which excludes the perturbation of bystander CD8+ T cells.

用于评估肿瘤发生过程中抗原特异性 CD8+ T 细胞动力学的引流淋巴结转移模型

Tumor antigen-specific CD8+ T cells from draining lymph nodes gain an accumulating importance in mounting anti-tumor immune response during tumorigenesis. ...

小鼠浅表淋巴结手术

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此视频中的章节

腹股沟淋巴结的活体显微镜

淋巴免疫治疗和疫苗接种小鼠

小鼠模型中上颈神经节切除术的手术技术

使用腹部皮下入路的改良腹腔镜腹股沟淋巴结清扫术

腹腔镜技术在猕猴肝和肠系膜淋巴结的串行采集

通过在小鼠中吸液性淋巴血管阻止淋巴流

小鼠下肢诱导继发性淋巴水肿的修订方法

用于评估肿瘤发生过程中抗原特异性 CD8⁺ T 细胞动力学的引流淋巴结转移模型

用于评估肿瘤发生过程中抗原特异性 CD8⁺ T 细胞动力学的引流淋巴结转移模型

用于评估肿瘤发生过程中抗原特异性 CD8⁺ T 细胞动力学的引流淋巴结转移模型