| In vitro Fluorescent Matrix Degradation Assay for Entamoeba histolytica
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Author:
Date:
2016-03-20
[Abstract] Fluorescent matrix degradation assay is a popular and widely used assay in the field of invadopodium biology (Artym et al., 2009). Matrix remodeling and degradation can be observed under both physiological and pathological conditions. Cancer cells extensively remodel and degrade the underlying matrix by employing actin-rich protrusive structures called invadosomes. Similar structures are formed by the protozoan parasite Entamoeba histolytica (E. histolytica), upon coming in contact with fibronectin, a major component of the host (extracellular matrix) ECM. Here, we describe a similar assay to measure matrix degradation by Entamoeba histolytica.
[摘要] 荧光基质降解测定是在invadopodium生物学领域中广泛使用的测定（Artym等人，2009）。 可以在生理和病理条件下观察基质重塑和降解。 癌细胞通过采用称为invadosomes的富含肌动蛋白的突出结构广泛地重塑和降解基础矩阵。 在与纤维连接蛋白（宿主（细胞外基质）ECM的主要组分）接触时，由原生动物寄生虫（即溶组织内阿米巴）（ histolytica ）形成类似的结构。 在这里，我们描述了通过溶组织内阿米巴测量基质降解的类似测定法。
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| Invadopodia Detection and Gelatin Degradation Assay
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Author:
Date:
2013-12-20
[Abstract] This protocol is designed to quantify invadopodia formation and activity. Invadopodia are protrusive structures elaborated by cancer cells that mediate cell attachment and remodeling of the extracellular matrix. These structures contribute to the ability of cancer cells to invade and metastasize. In this protocol, both the presence of invadopodia and their activity is simultaneously assessed and quantified by a fluorescent microscopy-based assay.
[摘要] 该协议旨在量化侵袭过程的形成和活性。 Invadopodia是介导细胞附着和细胞外基质重塑的癌细胞阐述的突出结构。 这些结构有助于癌细胞入侵和转移的能力。 在该方案中，通过基于荧光显微镜的测定法同时评估和定量侵袭多巴的存在及其活性。
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| Immunolabelling of Thin Slices of Mouse Descending Colon and Jejunum
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Author:
Date:
2013-10-20
[Abstract] This protocol describes a method for efficient immunolabelling of thin tissue slices containing a few rows of intact intestinal crypts, which yields large numbers of them being oriented favorably for recording stacks of optical sections aligned with the crypt long axis (Bellis et al., 2012). The latter can then be used for cell positional analysis, 3D-reconstruction and -analysis. The simple epithelium lining the small intestine is organized into contiguous crypts of Lieberkühn (Potten, 1998; Barker et al., 2012; De Mey and Freund, 2013) several of which making up a crypt/villus unit. Each crypt is a multicellular proliferation unit with a tight hierarchical organization. Under steady state conditions, the epithelium is continuously and rapidly renewed, driven by ...
[摘要] 该方案描述了一种用于有效免疫标记含有几行完整肠隐窝的薄组织切片的方法，其产生大量的定向以有利地记录与隐窝长轴对齐的光学部分的叠层（Bellis等人，2012） 。后者可用于细胞位置分析，3D重建和分析。小肠内衬的简单上皮组织为Lieberkühn（Potten，1998; Barker等，2012; De Mey和Freund，2013）的连续隐窝，其中几个构成了一个隐窝/绒毛单元。每个隐窝都是一个具有严密等级组织的多细胞增殖单元。在稳态条件下，上皮连续快速更新，由隐窝基底附近的多能肠SC分开，细胞从绒毛尖端移除。用于分析隐窝组织的技术在该领域起着重要的作用。通过使用从共聚焦扫描获得的光学部分和/或通过纵向隐窝轴的中心的诺瓦斯基光学器件获得最大效率，以将隐窝看作是从位于或靠近隐窝基底的细胞开始的分层谱系的两个细胞列。这使得能够对某些细胞能力进行位置分析，例如进行有丝分裂和细胞凋亡的DNA合成（Caldwell等人，2007; Fleming等，2007; Quyn等人，2010），对损伤作出反应（Potten等，1997 ）或表达基因（Barker等，2012; Bjerknes等，2012; ...
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