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Click-iT® EdU Alexa Fluor® 488 Imaging Kit

Click-iT ® EdU Alexa Fluor ® 488成像试剂盒

Company: Thermo Fisher Scientific
Catalog#: C10337
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Whole-mount Enteroid Proliferation Staining
[Abstract]  Small intestinal organoids, otherwise known as enteroids, have become an increasingly utilized model for intestinal biology in vitro as they recapitulate the various epithelial cells within the intestinal crypt (Mahe et al., 2013; Sato et al., 2009). Assessment of growth dynamics within these cultures is an important step to understanding how alterations in gene expression, treatment with protective and toxic agents, and genetic mutations alter properties essential for crypt growth and survival as well as the stem cell properties of the individual cells within the crypt. This protocol describes a method of visualization of proliferating cells within the crypt in three dimensions (Barrett et al., 2015). Whole-mount proliferation staining of enteroids ... [摘要]  小肠类器官,也称为肠袢,已经成为越来越多地用于肠道生物学的体外模型,因为它们重现了在肠隐窝内的各种上皮细胞(Mahe等人 ,2013; Sato et al。,2009)。在这些培养物中评估生长动力学是理解基因表达的改变,用保护性和毒性试剂处理以及遗传突变改变隐窝生长和存活所必需的性质以及细胞内单个细胞的干细胞性质的重要步骤地穴。该方案描述了在隐窝内三维的增殖细胞的可视化方法(Barrett等人,2015)。使用EdU掺入的肠衣的整体增殖染色使得研究者能够观察到肠内的所有增殖细胞,而不是如在包埋和切片中所见到的在薄切片中获得生长信息,确保来自干细胞区室的增殖的真实表现到隐窝的终末分化细胞。

EdU Based DNA Synthesis and Cell Proliferation Assay in Maize Infected by the Smut Fungus Ustilago maydis
[Abstract]  The basidiomycetous smut fungus Ustilago maydis (U. maydis) infects all aerial parts of its host plant maize (Zea mays L.). Infection is seen in the form of prominent tumorous symptoms after the establishment of a biotrophic interaction with the host, usually around 5-6 days after infection. The fungus colonizes the various developmentally distinct aerial organs at different stages of development. Formation of tumors is coupled with the induction of host cell division. Activation of cell division can be understood as a measure of DNA synthesis which is triggered to induce rapid divisions in host cell. This developed protocol helps in tracking tumor induction in U. maydis by monitoring of DNA synthesis in planta. Infected leaves were treated ... [摘要]  落叶青霉菌(Basidiomycetous smut fungus)Ustilago maydis( U。maydis )感染其宿主植物玉米的所有地上部分( Zea mays 感染在与宿主生物营养相互作用建立后通常在感染后约5-6天以突出的肿瘤症状的形式见到。真菌在不同发育阶段定殖各种发育不同的气生器官。肿瘤的形成与宿主细胞分裂的诱导相结合。细胞分裂的激活可以理解为DNA合成的量度,其被触发以诱导宿主细胞中的快速分裂。这种开发的协议有助于跟踪肿瘤的诱导。 maydis通过监测植物中的DNA合成来实现。 在幼苗叶中感染的几个阶段用5-乙炔基-2-脱氧尿苷(EdU)处理感染的叶并标记。通过连接荧光标签显现S期细胞中的EdU掺入,并用碘化丙啶(PI)染色未分裂的玉米核。这个协议有助于了解肿瘤的发展。 maydis 通过共聚焦激光扫描显微术(Kelliher和Walbot,2011; Redkar等人,2015)

Detection of the Cell Proliferation Zone in Leaves by Using EdU
[Abstract]  Visualization of nuclei in S-phase cells in tissues is important for not only cell cycle research but also developmental research because morphogenesis is usually achieved by a combination of cell proliferation and cell expansion. Recently, DNA labeling with 5-ethynyl-2′-deoxyuridine (EdU), which is an analog of thymidine, has been used to visualize nuclei in S-phase cells to assess the activity of cell proliferation during development of plants. EdU is efficiently incorporated into newly synthesized DNA, and detection of EdU is based on the covalent reaction between EdU and Alexa Fluor® dye, which is one of useful fluorescent dyes; this allows us to use mild conditions for the assay without any DNA denaturation. This method could be easily applicable, and, indeed, has been ... [摘要]  组织中S期细胞中核的可视化对于不仅细胞周期研究而且对发育研究是重要的,因为形态发生通常通过细胞增殖和细胞扩增的组合来实现。 最近,用5-乙炔基-2'-脱氧尿苷(EdU)(其是胸苷的类似物)的DNA标记已用于在S期细胞中显现细胞核以评估植物发育过程中细胞增殖的活性。 EdU有效地并入新合成的DNA中,并且EdU的检测基于EdU和作为有用的荧光染料之一的Alexa Fluor 染料之间的共价反应; 这允许我们使用温和条件进行测定而没有任何DNA变性。 该方法可以容易地应用,并且实际上已经用于各种模型和非模型植物物种。 在这里,我们已经描述了一个协议开发用于检测叶中S期细胞中的核。