| Analysis of Chromosome Condensation/Decondensation During Mitosis by EdU Incorporation in Nigella damascena L. Seedling Roots
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Author:
Date:
2018-02-05
[Abstract] To investigate the chromosome dynamics during mitosis, it is convenient to mark the discrete chromosome foci and then analyze their spatial rearrangements during prophase condensation and telophase decondensation. To label the chromosome regions in plant chromosomes, we incorporated the synthetic nucleotide, 5-ethynyl-2’-deoxyuridine (EdU), which can be detected by click-chemistry, into chromatin during replication. Here, we described a protocol of a method based on the application of semi-thin sections of Nigella damascena L. roots embedded in LR White acrylic resin. The thickness of semi-thin (100-250 nm) sections is significantly lower than that of optical sections even if a confocal microscope was used. This approach may also be suitable for work with any tissue fragments or ...
[摘要] 为了研究有丝分裂期间的染色体动力学，可以方便地标记离散的染色体焦点，然后分析它们在前期缩合和末期解聚期间的空间重排。 为了在植物染色体上标记染色体区域，我们将可以通过点击化学检测的合成核苷酸5-乙炔基-2'-脱氧尿苷（EdU）掺入染色质中。 在这里，我们描述了一种方法的协议，该方法基于嵌入在LR白色丙烯酸树脂中的
【背景】关于染色体组织的大多数数据是从少数模式生物的研究中获得的，其中大部分是哺乳动物。在具有大基因组的植物中，染色体显着大于迄今为止研究的动物染色体。为了研究有丝分裂期间的染色体动力学，有必要标记离散的染色体焦点，然后分析它们在前期凝结和末期解聚过程中的空间重排。为了标记染色体区域，我们引入了合成的核苷酸5-乙炔基-2'-脱氧尿苷（EdU）（Kuznetsova等人，2017）。 EdU的检测基于点击反应，其是叠氮化物和炔烃之间的铜催化反应。 EdU含有可与含叠氮化物的检测试剂反应的炔。
在S期后期并入EdU的细胞中可看到标记区域（即，分离的标记的点）的最合适的分布。短脉冲标记了所有S期细胞，并且EdU标记的有丝分裂图的初始外观因此表示在S期晚期标记的细胞穿过有丝分裂所需的时间。 ...
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| Immunoelectron Microscopy in Chlamydomonas Cells
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Author:
Date:
2013-03-05
[Abstract] The method of immunoelectron microscopy is intended for localization of proteins inside the cells of Chlamydomonas reinhardtii or other microalgae and cyanobacteria. This protocol was used to study localization of carbonic anhydrase Cah3 with antibodies raised in rabbit, though it can be used to localize any other abundant protein. Primary rabbit antibodies are recommended because they react quickly and specifically with proteins of C. reinhardtii. If primary antibodies other than rabbit are used, the blocking procedure and time of incubation with primary and secondary antibodies should be adjusted.
[摘要] 免疫电子显微镜法的目的是将蛋白质定位在莱茵衣藻或其它微藻和蓝细菌的细胞内。 该方案用于研究碳酸酐酶Cah3与在兔中产生的抗体的定位，尽管其可以用于定位任何其他丰富的蛋白质。 建议使用初级兔抗体，因为它们能快速和特异性地与C的蛋白反应。 reinhardtii 。 如果使用除兔以外的第一抗体，应当调整与第一和第二抗体孵育的封闭程序和时间。
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| Plastic Embedding and Sectioning of Plant Tissues
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Author:
Date:
2012-12-05
[Abstract] Plastic (resin) embedding provides exclusively improvements to cellular definition compared to paraffin embedding. The combination of strongly cross-linking paraformaldehyde with glutaraldehyde and post fixed with OsO4 is the fixative of choice for high-resolution light microscopy and electron microscopy. For this reason, this method is an ideal tool for visualizing plant cellular morphology and phenotype.
[摘要] 与石蜡包埋相比，塑料（树脂）包埋仅提供细胞定义。 强交联的多聚甲醛与戊二醛的组合以及用OsO 4后固定的组合是用于高分辨率光学显微镜和电子显微镜的固定剂。 因此，这种方法是可视化植物细胞形态和表型的理想工具。
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