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Trypan Blue Solution


Company: Thermo Fisher Scientific
Catalog#: 15250061
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Qualitative in vivo Bioluminescence Imaging
[Abstract]  Bioluminescence imaging (BLI) technology is an advanced method of carrying out molecular imaging on live laboratory animals in vivo. This powerful technique is widely-used in studying a variety of biological processes, and it has been an ideal tool in exploring tumor growth and metastatic spread in real-time. This technique ensures the optimal use of laboratory animal resources, particularly the ethical principle of reduction in animal use, given its non-invasive nature, ensuring that ongoing biological processes can be studied over time in the same animal, without the need to euthanize groups of mice at specific time points. In this protocol, the luciferase imaging technique was developed to study the effect of co-inoculating pericytes (contractile, αSMA+ mesenchymal ... [摘要]  生物发光成像(BLI)技术是一种在体内实验室动物上进行分子成像的先进方法。 这种强大的技术广泛应用于研究各种生物过程,是实时探索肿瘤生长和转移扩散的理想工具。 该技术确保实验室动物资源的最佳利用,特别是减少动物使用的伦理原则,考虑到其非侵入性,确保可以在同一动物中随时间研究正在进行的生物过程,而无需安乐死 小鼠在特定的时间点。 在该方案中,开发了荧光素酶成像技术以研究共同接种周细胞(收缩性,αSMA + 间充质干细胞样细胞,位于微血管内的细胞)对生长和转移性扩散的影响。 卵巢癌使用侵袭性卵巢癌细胞系-OVCAR-5-作为例子。

【背景】生物发光成像(BLI)的原理是基于相对简单的生化过程的发光特性,即,荧光素酶介导的分子底物荧光素氧化产生光。在癌症研究中,BLI是一种流行的工具(Contag et ...

Activation of Fibroblast Contractility via Cell-Cell Interactions and Soluble Signals
[Abstract]  The collagen contraction assay is an in vitro, three-dimensional method to determine the factor(s) affecting the contractile behavior of activated cells such as fibroblasts in either physiological or pathological scenarios. The collagen lattices/hydrogels are seeded with fibroblasts to mimic the interactions between these cells and their surrounding extracellular matrix proteins in the connective tissue. This method is an important platform to assess components as potential therapeutic targets to prevent pathologies such as fibrosis, which are manifestations of hyperactivated fibroblasts. We have described a basic version of this collagen contraction assay, which is amenable to customization using different cell types under diverse experimental conditions. [摘要]  胶原收缩测定是体外三维方法,用于确定影响生理或病理场景中活化细胞如成纤维细胞的收缩行为的因子。 胶原蛋白晶格/水凝胶用成纤维细胞接种,以模拟这些细胞与其周围细胞外基质蛋白在结缔组织中的相互作用。 该方法是评估组分作为潜在治疗靶标的重要平台,以预防纤维化等病症,这些病症是过度活化的成纤维细胞的表现。 我们已经描述了这种胶原收缩测定的基本版本,其适于在不同实验条件下使用不同细胞类型进行定制。

【背景】细胞外基质的组织收缩和重塑是许多生理条件(例如伤口愈合)中的基本过程。这两种现象的核心是成纤维细胞,它不仅产生和分泌细胞外基质蛋白,而且还可以通过机械相互作用重组它们。有趣的是,这些细胞行为通常在诸如纤维化的病理条件下被夸大(Desmoulière et al。,2005),从而说明需要理解这些过程的分子调节。虽然人们早就知道,胶原蛋白是细胞外基质的主要成分之一,是组织收缩的主要参与者(Bell et al。,1979),对机械细节的透彻理解。这个过程仍然难以捉摸。对体外成纤维细胞胶原基质体外收缩的研究使研究人员能够识别导致组织收缩的新型运动员(Ngo et al。,2006; Su and Chen, 2015年)。基于该测定,可溶性因子如TGFβ(Levi-Schaffer 等,1999)和免疫细胞(Garbuzenko et al。,2002; ...

Measurement of TLR4 and CD14 Receptor Endocytosis Using Flow Cytometry
[Abstract]  After recognizing extracellular bacterial lipopolysaccharide (LPS), the toll-like receptor 4 (TLR4)-CD14 signaling complex initiates two distinct signaling pathways–one from the plasma membrane and the other from the signaling endosomes (Kagan et al., 2008). Understanding the early stages of TLR4 signal transduction therefore requires a robust and quantitative method to measure LPS-triggered TLR4 and CD14 receptor endocytosis, one of the earliest events of LPS detection. Here, we describe a flow cytometry-based method that we used recently to study the role of the ion channel TRPM7 in TLR4 endocytosis (Schappe et al., 2018). The assay relies on stimulating the cells with LPS and measuring the cell surface levels of TLR4 (or CD14) at various time points using flow ... [摘要]  在识别细胞外细菌脂多糖(LPS)后,Toll样受体4(TLR4)-CD14信号传导复合物启动两种不同的信号传导途径 - 一种来自质膜,另一种来自信号传导内体(Kagan 等。,2008)。因此,了解TLR4信号转导的早期阶段需要一种稳健且定量的方法来测量LPS触发的TLR4和CD14受体内吞作用,这是LPS检测中最早发生的事件之一。在这里,我们描述了一种基于流式细胞术的方法,我们最近用它来研究离子通道TRPM7在TLR4内吞作用中的作用(Schappe et al。,2018)。该测定依赖于用LPS刺激细胞并使用流式细胞术在不同时间点测量TLR4(或CD14)的细胞表面水平。尽管我们详细描述了来自鼠骨髓来源的巨噬细胞的TLR4和CD14的方法,但它可以很容易地适应于在各种其他信号传导环境中评估受体内吞作用。

【背景】先天免疫细胞,包括巨噬细胞和树突细胞,使用各种模式识别受体(PRR)来调查其环境中的危险和病原体相关分子模式。来自各种亚细胞区室的PRR的贩运和信号传导实现了更广泛的免疫监视,并且已成为先天免疫的重要设计原则(Brubaker et al。,2015)。细菌内毒素LPS的检测高度依赖于TLR4及其共同受体CD14。 TLR4复合物的内吞作用需要CD14,并且对于LPS诱导的巨噬细胞活化是必需的(Zanoni 等人,2011; Tan ...