Name: immunostaining to visualize murine enteric nervous system development
Uploaded: 2015-04-29T13:00:00.000+00:00
Duration: 7 min 54 s
Description: Watch this Scientific Journal Video about Immunostaining to Visualize Murine Enteric Nervous System Development at JoVE.com

这项工作是支持的美国bythe小儿外科协会基金会奖（AG）和健康K08DK098271全国学院（AG）。

威斯康星州的动物护理和使用委员会的批准大学的所有程序。 1.准备解决方案<ol><li>使用1×磷酸盐缓冲盐水（PBS），为解剖缓冲液和冲洗液。 </li><li>由30克蔗糖和地点到瓶中制备30％的蔗糖。添加99毫升1×PBS中并加入1毫升10％的叠氮化钠。调匀，直到所有的蔗糖溶解。直到需要保存在4℃。 </li><li>制备通过混合的1×PBS，3％牛血清白蛋白（BSA）和0.1％的Triton-X-100的封闭溶液。调匀，直到需要在冰箱里储存。 </li><li>制备在1×PBS中的8％的多聚甲醛（PFA）溶液通过称重的PFA的适量的1×PBS中，然后孵育在65℃直到它完全溶解。商店在25毫升等份在-20℃冷冻，直到需要。稀释至4％PFA中的1×PBS在使用的当天。 </li></ol> 2.胚胎和肠道解剖离子<ol><li>按照机构动物护理和使用委员会批准的方案，安乐死定时怀孕小鼠和子宫转移到60毫米培养皿中含有冰冷的1×PBS。 </li><li>在使用锋利的剪刀解剖镜下，切开子宫壁并暴露出胚胎。除去从子宫和地点胚胎成含冰冷的1×PBS中的清洁60毫米培养皿。 </li><li>安乐死在冰冷的1X PBS每个胚胎被斩首。如果您使用的是含有荧光蛋白转基因小鼠，在荧光照明，标识正转基因胚胎。 </li><li>解剖使用解剖显微镜每个胚GI道。用细镊子，定向，使得所述左侧朝上，右侧是针对培养皿底部的胚胎。从胚胎中取出上体壁以暴露内部器官。将背体壁和内脏之间的细镊子。铬oss骨粉对对方的镊子在剪刀状切割动作，从胚胎取出内脏。 </li><li>从周围器官进一步分解剖GI道远，然后将每个GI道到含有冰冷的1×PBS 1.5ml微量离心管中。 </li></ol> 3.固定GI大片<ol><li>冲洗每个GI道3次，用冰冷的1×PBS，然后替换用4％PFA。固定在摇摆平台在RT 1.5小时在1.5 ml微量离心管在胃肠道。冲洗胃肠道3次，每次5分钟，RT，然后进行1小时的摇动平台上。在此阶段，存储在胃肠道，在4℃下在30％蔗糖中含有1x PBS中的0.1％的叠氮化钠，直到需要。 注意:可替换地，存储在30％蔗糖的胚胎为长达一年而不对组织的完整性的任何影响。在30％的蔗糖样品的存储允许的样品无论是对免疫染色或进入十月为低温sectioni的购买处理NG。另外，继续进行下面详细介绍的免疫协议。 </li></ol> 4.免疫染色协议<ol><li>如果样本已储存在30％的蔗糖，冲洗他们在摇摆平台上在1×PBS中3次20分钟。 </li><li>放置在胃肠道进入封闭溶液在摇摆平台上1小时，在室温。 </li><li>除去封闭液孵育胃肠道与稀释于封闭为任一4小时，在室温或O / N在4℃下在摇摆平台上的解决方案的主要抗体的适当的量。使用1:1000稀释的人抗胡抗体（血清自患者获得），1:1000稀释的鸡抗 - 绿色荧光蛋白（GFP）的抗体和1:100稀释的山羊抗胆碱抗体。 注意:我们利用了从患者局部获得的人抗胡抗体，然而，抗胡抗体是可商购的，例如，小鼠抗胡，（使用1:500稀释）。 </li><li>在冲洗1X PBS胃肠道3次，每次5分钟，然后进行1小时，在室温在摇摆平台上。 </li><li>在封闭溶液在摇摆平台上为4小时，在室温或O / N在4℃下500:取代的1×PBS稀释1的二次抗体。使用驴抗人胺反应性染料如Dylight，驴抗鸡的Cy2和驴抗山羊Cy5的。如果使用小鼠抗胡抗体然后用1:500稀释的驴抗小鼠胺反应性染料405。 注:内源性GFP和tdTomato表达和免疫染色随发育年龄的清晰度的强度。我们通常免疫染色胚胎胆单独在0.2ml管中加入150μl的染色溶液，以减少所使用的抗体的量，同时确保组织的有效染色。 </li><li>冲洗在1×PBS中的3倍胃肠道5分钟，然后进行1小时，在室温在摇摆平台上。 </li><li>放置几滴荧光封固剂用DAPI的载玻片上。沉浸GI TRA克拉到荧光封固剂并直接在组织上添加一个玻璃盖。荧光封固剂-G是一种水溶性化合物，它提供了半永久性密封。 注:使用荧光安装介质如的Vectashield这是基于安装介质，防止褪色和抗体的漂白甘油。这两种产品使组织被存储的时间，在4℃下长期使用。 </li><li>捕捉利用共聚焦显微镜荧光每个人的图片。使用激发波长为荧光团和过滤器使用的表3中。 </li><li>使用根据用于捕捉图像共焦的类型适当的软件执行计算机辅助图像分析。 </li></ol>

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The authors have nothing to disclose.

我们的实验室和其他人已经表明，在HSCR肠缺陷不限于无神经节结肠但近侧延伸，连成神经节小肠5,15,16。这些改变包括改变在ENS神经元密度和神经递质的表型，并且可以考虑动力障碍已被观察到的患者HSCR。我们利用上述技术，我们在努力了解ENS形成的决定因素。具体地，这些技术已被用来可视化神经元一氧化氮合成酶（nNOS的）神经元，血管活性肠肽神经元（VIP），以及胆碱乙酰转移酶（C...

一个发挥作用的肠神经系统（ENS），控制运动，营养吸收和局部血流量，是生活中必不可少的1。该ENS是由神经嵴细胞（NCC）的增殖，迁移和定居的肠道，在那里他们分化成神经节含有神经元和神经胶质细胞形成。先天性巨结肠症（文中HSCR，在线孟德尔遗传），一个multigeneic先天性疾病与1 4000活产儿发病，可以被认为是原型病研究打乱ENS形成。在HSCR，NCC无法迁移并定居于远端肠2的可变长度。此外，其他常见的胃肠道（GI）中的儿科人群发育缺陷，如肛门直肠畸形，肠atresias，和运动障碍是与在基本ENS功能紊乱有关，并有可能与微妙，低估，解剖变化和功能性变化有关在ENS 3-6。因此，技术，使我们能够理解的ENS形成发展的决定因素可能会阐明其发病机制及小儿胃肠道疾病的潜在治疗轻。 以下迁移和殖民化，NCC分化为神经元特异性神经递质的表型标记。胆碱能神经元包括肠神经元7的约60％，并且可以通过染色胆碱乙酰转移酶（ChAT的），合成酶的兴奋性神经递质乙酰胆碱来检测。在历史上，试图可视胆碱能神经元是由不同的针对中枢神经系统的抗体的抗原特异性混淆（CNS）的ChAT的与外周神经系统（PNS）的ChAT的8-10。然而，针对胎盘胆碱抗体识别中枢和外周胆碱11-13，和我们最近所描述的技术，允许visuaENS胆碱能神经元的高灵敏度补肾中药较早发展比已取得的ChAT记者线14。 在这里，我们提出了解剖，固定小鼠胚胎胃肠道和免疫可视化ENS神经递质表达的神经元的技术。在这些研究中，我们已经利用交配R26R聊天的Cre小鼠:floxSTOP:tdTomato动物生产聊天的Cre; R26R:floxSTOP:tdTomato小鼠（整个手稿定义 ​​为聊天的Cre tdTomato）。这些动物，然后用纯合子聊天GFP报告小鼠交配，获得表达的小鼠都来检测胆碱表达荧光14记者。这两个记者动物是在C57BL / 6J背景和市售（Jackson实验室，巴港，缅因州）。

<table><tbody><tr><td>Phosphate buffered saline</td><td>Oxoid</td><td>BR0014G</td><td></td></tr><tr><td>Sucrose</td><td>Fisher</td><td>S2</td><td></td></tr><tr><td>Sodium azide</td><td>Fisher</td><td>BP9221</td><td></td></tr><tr><td>Bovine serum albumin</td><td>Fisher</td><td>BP1605</td><td></td></tr><tr><td>Triton X-100</td><td>Sigma</td><td>X100</td><td></td></tr><tr><td>Paraformaldehyde</td><td>Sigma</td><td>158127</td><td></td></tr><tr><td>60 mm Petri dishes</td><td>Fisher</td><td>FB0875713A</td><td></td></tr><tr><td>Fluorescence microscope</td><td>Nikon</td><td>SMZ-18 stereoscope</td><td></td></tr><tr><td>Dissection microscope</td><td>Nikon</td><td>SMZ-18 stereoscope</td><td></td></tr><tr><td>Fine forceps</td><td>Fine science tools</td><td>11252-20</td><td></td></tr><tr><td>1.5 ml Eppendorf tubes</td><td>VWR</td><td>20170-038</td><td></td></tr><tr><td>Fluoromount-G</td><td>SouthernBiotech, Birmingham, AL</td><td>0100-01</td><td></td></tr><tr><td>Glass slides</td><td>Fisher</td><td>12-550-15</td><td></td></tr><tr><td>Cover glass</td><td>VWR</td><td>16004-330</td><td></td></tr><tr><td>Confocal microscope</td><td>Nikon</td><td>Nikon A1</td><td></td></tr><tr><td>Nikon Elements</td><td>Nikon</td><td></td><td></td></tr></tbody></table>

immunostaining to visualize murine enteric nervous system development

The enteric nervous system is formed by neural crest cells that proliferate, migrate and colonize the gut. Following colonization, neural crest cells must then differentiate into neurons with markers specific for their neurotransmitter phenotype. Cholinergic neurons, a major neurotransmitter phenotype in the enteric nervous system, are identified by staining for choline acetyltransferase (ChAT), the synthesizing enzyme for acetylcholine. Historical efforts to visualize cholinergic neurons have been hampered by antibodies with differing specificities to central nervous system versus peripheral nervous system ChAT. We and others have overcome this limitation by using an antibody against placental ChAT, which recognizes both central and peripheral ChAT, to successfully visualize embryonic enteric cholinergic neurons. Additionally, we have compared this antibody to genetic reporters for ChAT and shown that the antibody is more reliable during embryogenesis. This protocol describes a technique for dissecting, fixing and immunostaining of the murine embryonic gastrointestinal tract to visualize enteric nervous system neurotransmitter expression.

我们先前描述了小鼠表达GFP和tdTomato荧光记者，检测ChAT的表达14的产生。简单地说， 聊天的Cre小鼠交配R26R:floxSTOP:tdTomato动物生产聊天的Cre; R26R:floxSTOP:tdTomato小鼠（称为聊天的Cre tdTomato）。这些动物处死后交配的纯合的ChAT-GFP报道小鼠。胚胎中分离和组织之前被固定进行解剖并如上述进行免疫染色，并在表1中列出的抗体。远端小肠（SI）和近端...

Watch this Scientific Journal Video about Immunostaining to Visualize Murine Enteric Nervous System Development at JoVE.com

免疫染色可视化小鼠肠神经系统发育发育生物学|视频

Immunostaining to Visualize Murine Enteric Nervous System Development

The enteric nervous system is formed by neural crest cells that proliferate, migrate and colonize the gut. Neural crest cells differentiate into neurons with markers specific for their neurotransmitter phenotype. This protocol describes a technique for dissecting, fixing and immunostaining of the murine embryonic gastrointestinal tract to visualize enteric nervous system neurotransmitter expression.

免疫组化可视化小鼠肠神经系统的发展

The enteric nervous system is formed by neural crest cells that proliferate, migrate and colonize the gut. Following colonization, neural crest cells must ...

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11.3K 次观看.  University of Wisconsin School of Medicine and Public Health. The enteric nervous system is formed by neural crest cells that proliferate, migrate and colonize the gut. Neural crest cells differentiate into neurons with markers specific for their neurotransmitter phenotype. This protocol describes a technique for dissecting, fixing and immunostaining of the murine embryonic gastrointestinal tract to visualize enteric nervous system neurotransmitter expression.

视频: Immunostaining to Visualize Murine Enteric Nervous System Development

Multimodal Imaging of Stem Cell Implantation in the Central Nervous System of Mice

During the past decade, stem cell transplantation has gained increasing interest as primary or secondary therapeutic modality for a variety of diseases, both in preclinical and clinical studies. However, to date results regarding functional outcome and/or tissue regeneration following stem cell transplantation are quite diverse. Generally, a clinical benefit is observed without profound understanding of the underlying mechanism(s)1. Therefore, multiple efforts have led to the development of different molecular imaging modalities to monitor stem cell grafting with the ultimate aim to accurately evaluate survival, fate and physiology of grafted stem cells and/or their micro-environment. Changes observed in one or more parameters determined by molecular imaging might be related to the observed clinical effect. In this context, our studies focus on the combined use of bioluminescence imaging (BLI), magnetic resonance imaging (MRI) and histological analysis to evaluate stem cell grafting.
BLI is commonly used to non-invasively perform cell tracking and monitor cell survival in time following transplantation2-7, based on a biochemical reaction where cells expressing the Luciferase-reporter gene are able to emit light following interaction with its substrate (e.g. D-luciferin)8, 9. MRI on the other hand is a non-invasive technique which is clinically applicable10 and can be used to precisely locate cellular grafts with very high resolution11-15, although its sensitivity highly depends on the contrast generated after cell labeling with an MRI contrast agent. Finally, post-mortem histological analysis is the method of choice to validate research results obtained with non-invasive techniques with highest resolution and sensitivity. Moreover end-point histological analysis allows us to perform detailed phenotypic analysis of grafted cells and/or the surrounding tissue, based on the use of fluorescent reporter proteins and/or direct cell labeling with specific antibodies.
In summary, we here visually demonstrate the complementarities of BLI, MRI and histology to unravel different stem cell- and/or environment-associated characteristics following stem cell grafting in the CNS of mice. As an example, bone marrow-derived stromal cells, genetically engineered to express the enhanced Green Fluorescent Protein (eGFP) and firefly Luciferase (fLuc), and labeled with blue fluorescent micron-sized iron oxide particles (MPIOs), will be grafted in the CNS of immune-competent mice and outcome will be monitored by BLI, MRI and histology (Figure 1).

This article describes an optimized sequence of events for multimodal imaging of cellular grafts in rodent brain using: (i) in vivo bioluminescence and magnetic resonance imaging, and (ii) post mortem histological analysis. Combining these imaging modalities on a single animal allows cellular graft evaluation with high resolution, sensitivity and specificity.

在中央小鼠中枢神经系统的干细胞移植的多模态成像

During the past decade, stem cell transplantation has gained increasing interest as primary or secondary therapeutic modality for a variety of diseases, ...

科研

神经科学

Whole Mount Immunofluorescent Staining of the Neonatal Mouse Retina to Investigate Angiogenesis In vivo

Angiogenesis is the complex process of new blood vessel formation defined by the sprouting of new blood vessels from a pre-existing vessel network. Angiogenesis plays a key role not only in normal development of organs and tissues, but also in many diseases in which blood vessel formation is dysregulated, such as cancer, blindness and ischemic diseases. In adult life, blood vessels are generally quiescent so angiogenesis is an important target for novel drug development to try and regulate new vessel formation specifically in disease. In order to better understand angiogenesis and to develop appropriate strategies to regulate it, models are required that accurately reflect the different biological steps that are involved. The mouse neonatal retina provides an excellent model of angiogenesis because arteries, veins and capillaries develop to form a vascular plexus during the first week after birth. This model also has the advantage of having a two-dimensional (2D) structure making analysis straightforward compared with the complex 3D anatomy of other vascular networks. By analyzing the retinal vascular plexus at different times after birth, it is possible to observe the various stages of angiogenesis under the microscope. This article demonstrates a straightforward procedure for analyzing the vasculature of a mouse retina using fluorescent staining with isolectin and vascular specific antibodies.

Whole Mount Immunofluorescent Staining of the Neonatal Mouse Retina to Investigate Angiogenesis In vivo

The neonatal murine retina provides a well characterized physiological model of angiogenesis, which permits investigations of the roles of different genes or drugs that modulate angiogenesis in an in vivo context. Immunofluorescent staining to accurately visualize the vascular plexus is pivotal to the success of these types of studies.

整装免疫荧光染色新生小鼠视网膜血管生成调查<em&gt;在体内</em

Angiogenesis  is the complex process of new blood vessel formation defined by the sprouting  of new blood vessels from a pre-existing vessel network. ...

An In-vitro Preparation of Isolated Enteric Neurons and Glia from the Myenteric Plexus of the Adult Mouse

The enteric nervous system is a vast network of neurons and glia running the length of the gastrointestinal tract that functionally controls gastrointestinal motility. A procedure for the isolation and culture of a mixed population of neurons and glia from the myenteric plexus is described. The primary cultures can be maintained for over 7 days, with connections developing among the neurons and glia. The longitudinal muscle strip with the attached myenteric plexus is stripped from the underlying circular muscle of the mouse ileum or colon and subjected to enzymatic digestion. In sterile conditions, the isolated neuronal and glia population are preserved within the pellet following centrifugation and plated on coverslips. Within 24-48 hr, neurite outgrowth occurs and neurons can be identified by pan-neuronal markers. After two days in culture, isolated neurons fire action potentials as observed by patch clamp studies. Furthermore, enteric glia can also be identified by GFAP staining. A network of neurons and glia in close apposition forms within 5 - 7 days. Enteric neurons can be individually and directly studied using methods such as immunohistochemistry, electrophysiology, calcium imaging, and single-cell PCR. Furthermore, this procedure can be performed in genetically modified animals. This methodology is simple to perform and inexpensive. Overall, this protocol exposes the components of the enteric nervous system in an easily manipulated manner so that we may better discover the functionality of the ENS in normal and disease states.

An In-vitro Preparation of Isolated Enteric Neurons and Glia from the Myenteric Plexus of the Adult Mouse

We demonstrate a cell culture protocol for the direct study of neuronal and glial components of the enteric nervous system. A neuron/glia mixed culture on coverslips is prepared from the myenteric plexus of adult mouse providing the ability to examine individual neuron and glia function by electrophysiology, immunohistochemical, etc.

一个<em在体外</em从成年小鼠的肌间神经丛制备隔离肠道神经元和神经胶质

The enteric nervous system is a vast network of neurons and glia running the length of the gastrointestinal tract that functionally controls ...

A Quantitative Cell Migration Assay for Murine Enteric Neural Progenitors

Neural crest cells (NCC) are a transient and multipotent cell population that originates from the dorsal neural tube and migrates extensively throughout the developing vertebrate embryo. In addition to providing peripheral glia and neurons, NCC generate melanocytes as well as most of the cranio-facial skeleton. NCC migration and differentiation is controlled by a combination of their axial origin along the neural tube and their exposure to regionally distinct extracellular cues. Such contribution of extracellular ligands is especially evident during the formation of the enteric nervous system (ENS), a complex interconnected network of neural ganglia that locally controls (among other things) gut muscle movement and intestinal motility. Most of the ENS is derived from a small initial pool of NCC that undertake a long journey in order to colonize - in a rostral to caudal fashion - the entire length of the prospective gut. Among several signaling pathways known to influence enteric NCC colonization, GDNF/RET signaling is recognized as the most important. Indeed, spatiotemporally controlled secretion of the RET ligand GDNF by the gut mesenchyme is chiefly responsible for the attraction and guidance of RET-expressing enteric NCC to and within the embryonic gut. Here, we describe an ex vivo cell migration assay, making use of a transgenic mouse line possessing fluorescently labeled NCC, which allows precise quantification of enteric NCC migration potential in the presence of various growth factors, including GDNF.

We present an ex vivo cell migration assay that allows precise quantification of enteric neural crest cell migration potential in the presence of various growth factors.

定量细胞迁移检测方法的鼠肠神经前体细胞

Neural crest cells (NCC) are a transient and multipotent cell population that originates from the dorsal neural tube and migrates extensively throughout ...

In Situ Ca2+ Imaging of the Enteric Nervous System

Reflex behaviors of the intestine are controlled by the enteric nervous system (ENS). The ENS is an integrative network of neurons and glia in two ganglionated plexuses housed in the gut wall. Enteric neurons and enteric glia are the only cell types within the enteric ganglia. The activity of enteric neurons and glia is responsible for coordinating intestinal functions. This protocol describes methods for observing the activity of neurons and glia within the intact ENS by imaging intracellular calcium (Ca2+) transients with fluorescent indicator dyes. Our technical discussion focuses on methods for Ca2+ imaging in whole-mount preparations of the myenteric plexus from the rodent bowel. Bulk loading of ENS whole-mounts with a high-affinity Ca2+ indicator such as Fluo-4 permits measurements of Ca2+ responses in individual neurons or glial cells. These responses can be evoked repeatedly and reliably, which permits quantitative studies using pharmacological tools. Ca2+ responses in cells of the ENS are recorded using a fluorescence microscope equipped with a cooled charge-coupled device (CCD) camera. Fluorescence measurements obtained using Ca2+ imaging in whole-mount preparations offer a straightforward means of characterizing the mechanisms and potential functional consequences of Ca2+ responses in enteric neurons and glial cells.

In Situ Ca2+ Imaging of the Enteric Nervous System

The enteric nervous system (ENS) is a network of neurons and glia located in the gut wall that controls intestinal reflexes. This protocol describes methods for recording the activity of enteric neurons and glia in live preparations of ENS using Ca2+ imaging.

在肠神经系统的原位 钙成像

Reflex behaviors of the intestine are controlled by the enteric nervous system (ENS). The ENS is an integrative network of neurons and glia in two ...

胚胎小鼠脑混合细胞培养物研究感染和先天免疫

Establishing Mixed Neuronal and Glial Cell Cultures from Embryonic Mouse Brains to Study Infection and Innate Immunity

Models of the central nervous system (CNS) must recapitulate the complex network of interconnected cells found in vivo. The CNS consists primarily of neurons, astrocytes, oligodendrocytes, and microglia. Due to increasing efforts to replace and reduce animal use, a variety of in vitro cell culture systems have been developed to explore innate cell properties, which allow the development of therapeutics for CNS infections and pathologies. Whilst certain research questions can be addressed by human-based cell culture systems, such as (induced) pluripotent stem cells, working with human cells has its own limitations with regard to availability, costs, and ethics. Here, we describe a unique protocol for isolating and culturing cells from embryonic mouse brains. The resulting mixed neural cell cultures mimic several cell populations and interactions found in the brain in vivo. Compared to current equivalent methods, this protocol more closely mimics the characteristics of the brain and also garners more cells, thus allowing for more experimental conditions to be investigated from one pregnant mouse. Further, the protocol is relatively easy and highly reproducible. These cultures have been optimized for use at various scales, including 96-well based high throughput screens, 24-well microscopy analysis, and 6-well cultures for flow cytometry and reverse transcription-quantitative polymerase chain reaction (RT-qPCR) analysis. This culture method is a powerful tool to investigate infection and immunity within the context of some of the complexity of the CNS with the convenience of in vitro methods.

作者聚焦:从胚胎小鼠大脑中建立混合神经元和神经胶质细胞培养物以研究感染和先天免疫  

This protocol presents a unique way of generating central nervous system cell cultures from embryonic day 17 mouse brains for neuro(immuno)logy research. This model can be analyzed using various experimental techniques, including RT-qPCR, microscopy, ELISA, and flow cytometry.

从胚胎小鼠大脑中建立混合神经元和神经胶质细胞培养物以研究感染和先天免疫

Models of the central nervous system (CNS) must recapitulate the complex network of interconnected cells found in vivo. The CNS consists primarily of ...

Using a Whole-mount Immunohistochemical Method to Study the Innervation of the Biliary Tract in Suncus murinus

This work describes the whole-mount immunohistochemistry staining method in detail, using neurofilament protein antibody to label the innervation of the biliary tract in Suncus murinus (S. murinus&#160;). First, the specimen was dissected from S. murinus and fixed in 4% paraformaldehyde (PFA). Second, an enzymatic treatment and potential endogenous peroxidase inactivation were performed. The specimen was then exposed to the primary antibody, anti-neurofilament protein antibody, for 3-6 days. It was then incubated with the secondary antibody conjugated with horseradish peroxidase. The color reaction was revealed by reacting the specimen with a 3,3&#39;-diaminobenzidine (DAB) substrate. This method can be applied to analyze the innervation of all visceral organs. Furthermore, this protocol can also be adapted to test other neuronal antibodies, but optimization of the antibodies should be done first. This method was originally introduced by Kuratani and Tanaka1,2,3.

Using a Whole-mount Immunohistochemical Method to Study the Innervation of the Biliary Tract in Suncus murinus

A whole-mount immunohistochemical approach, to visualize neurofilament protein expression in the extrahepatic biliary tract in Suncus murinus. is presented here. This protocol can be used to analyze the innervation of all visceral organs in S. murinus or other species.

使用全身免疫组织化学方法研究胆道的神经支配<em&gt; Suncus murinus</em

This work describes the whole-mount immunohistochemistry staining method in detail, using neurofilament protein antibody to label the innervation of the ...

生物学

Immunofluorescence Staining for Visualizing Proliferated Neural Stem Cells in Mouse Brain Sections

Source: Quintanilla, L. J., et al. <a href="https://app-jove-com.bdigitaluss.remotexs.co/v/59237">Assaying Circuit Specific Regulation of Adult Hippocampal Neural Precursor Cells.</a> J. Vis. Exp. (2019)
This video demonstrates the procedure for staining newly proliferated neural stem cells in brain tissue sections using EdU incorporation, followed by immunofluorescence to visualize specific proteins, enhancing the understanding of cell dynamics and proliferation.

免疫荧光染色用于观察小鼠脑切片中增殖的神经干细胞

All procedures involving animal samples have been reviewed and approved by the appropriate animal ethical review committee.
1. Immunohistochemistry

	...

免疫组化可视化小鼠肠神经系统的发展

摘要

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在中央小鼠中枢神经系统的干细胞移植的多模态成像

整装免疫荧光染色新生小鼠视网膜血管生成调查

一个

定量细胞迁移检测方法的鼠肠神经前体细胞

在肠神经系统的原位 ^钙成像

从胚胎小鼠大脑中建立混合神经元和神经胶质细胞培养物以研究感染和先天免疫

从胚胎小鼠大脑中建立混合神经元和神经胶质细胞培养物以研究感染和先天免疫

从胚胎小鼠大脑中建立混合神经元和神经胶质细胞培养物以研究感染和先天免疫

使用全身免疫组织化学方法研究胆道的神经支配

免疫荧光染色用于观察小鼠脑切片中增殖的神经干细胞

免疫组化可视化小鼠肠神经系统的发展

摘要

探索更多视频

此视频中的章节

在中央小鼠中枢神经系统的干细胞移植的多模态成像

整装免疫荧光染色新生小鼠视网膜血管生成调查

一个

定量细胞迁移检测方法的鼠肠神经前体细胞

在肠神经系统的原位 钙成像

从胚胎小鼠大脑中建立混合神经元和神经胶质细胞培养物以研究感染和先天免疫

从胚胎小鼠大脑中建立混合神经元和神经胶质细胞培养物以研究感染和先天免疫

从胚胎小鼠大脑中建立混合神经元和神经胶质细胞培养物以研究感染和先天免疫

使用全身免疫组织化学方法研究胆道的神经支配

免疫荧光染色用于观察小鼠脑切片中增殖的神经干细胞

在肠神经系统的原位 ^钙成像