| Using xCELLigence RTCA Instrument to Measure Cell Adhesion
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Author:
Date:
2017-12-20
[Abstract] Cell adhesion to neighbouring cells and to the underlying extracellular matrix (ECM) is a fundamental requirement for the existence of multicellular organisms. As such, the formation, stability and dissociation of cell adhesions are subject to tight control in space and time and perturbations within the sophisticated adhesion machinery are associated with a variety of human pathologies. Here, we outline a simple protocol to monitor alterations in cell adhesion to the ECM, for example, following genetic manipulations or overexpression of a protein of interest or in response to drug treatment, ...
[摘要] 细胞与相邻细胞和基础细胞外基质（ECM）的粘附是多细胞生物存在的基本要求。 因此，细胞粘附的形成，稳定和解离在时间和空间上受到严格的控制，复杂的粘附机制内的干扰与各种人类病理有关。 在这里，我们概述了一个简单的协议，以监测细胞粘附到ECM的变化，例如，在遗传操作或目标蛋白的过表达或响应药物治疗后，使用xCELLigence实时细胞分析（RTCA）系统。
【背景】负责细胞粘附到下面的ECM的主要分子是称为整联蛋白的跨膜异二聚体受体家族。整合素的激活和与ECM的结合触发向整合素胞质尾部募集大量信号传导，支架和细胞骨架蛋白。总之，这些粘附成分代表了负责调节许多重要细胞过程（包括细胞增殖，存活，迁移和分化）的复杂且高度动态的机制。与维持正常生理功能的重要角色一致，整合素介导的粘附和信号传导失调是许多人类疾病（包括出血性疾病，心血管疾病和癌症）发病的先导（Giancotti and Ruoslahti，1999;Bökeland Brown，2002; Huveneers and Danen，2009; Legate等人，2009; Bouvard等人，2013; Calderwood等人，2013; Horton等人，等，2015; Seguin等，，2015）。因此，整合素依赖性细胞 - 细胞外基质粘附的研究是一个研究热点，也是许多生物学领域广泛关注的课题。 ...
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| Synaptoneurosome Preparation from C57BL/6 Striata
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Author:
Date:
2016-02-20
[Abstract] Activity-dependent local mRNA translation endows synapses to remodel their structure and function (Bramham and Wells, 2007). This process is tightly controlled by the state of phosphorylation of several components of the translational machinery including initiation factors and ribosomal proteins (Buffington et al., 2014). The present protocol describes a method to prepare striatal synaptoneurosomes, from adult mice, containing both pre- and postsynaptic elements in which the level of synaptic phospho-proteins can be quantified (Biever et al., 2015).
[摘要] 活动依赖的局部mRNA翻译赋予突触重塑其结构和功能（Bramham和Wells，2007）。 该过程由包括起始因子和核糖体蛋白在内的翻译机理的几个组分的磷酸化状态来严格控制（Buffington等，2014）。 本方案描述了一种从成年小鼠制备纹状体synaptoneurosomes的方法，其含有可以量化突触磷酸蛋白水平的突触前和突触后元件（Biever等，2015）。
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| Protocol for T-cell Adhesion Strength on Tumor Cells under Flow Conditions
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Author:
Date:
2013-10-20
[Abstract] This method allows evaluating the relative adhesion strength between T lymphocytes and specific adherent target cells using a shear force in flow chambers. It is based on the measure of the resistance of conjugates formed between T cells and adherent tumor cells to shear stress in a microfluidic system. For this purpose, T cells, stained with a CellTracker probe, are added into flow channels containing a monolayer of adherent target cells and their progressive detachment under a constant shear stress is then recorded using a fluorescent microscope.
[摘要] 该方法允许使用流动室中的剪切力评估T淋巴细胞和特异性粘附靶细胞之间的相对粘附强度。 其基于在微流体系统中T细胞和粘附的肿瘤细胞之间形成的缀合物对剪切应力的抗性的测量。 为此，将用CellTracker探针染色的T细胞加入含有单层粘附靶细胞的流动通道中，然后使用荧光显微镜记录在恒定剪切应力下的其逐渐分离。
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