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Author:
Date:
2020-12-05
[Abstract] During immune responses, B cells home to lymph nodes (LN), where they encounter antigens. Homing starts with capture and L-selectin-dependent rolling on the activated endothelium of high endothelial venules (HEV). After recognition of chemokines presented on HEV, activation of B cell integrins occurs mediating firm arrest. Subsequently, B cells crawl to the spot of extravasation to enter the LN. Extravasation can be visualized and quantified in vivo by intravital microscopy (IVM) of the inguinal LN. Here, we describe an established protocol that permits detailed in vivo analysis of B cell recruitment to LN under sterile inflammatory conditions. We describe data acquisition, exportation, quantification, and statistical analysis using specialized software. IVM of LN is a powerful technique ...
[摘要] [摘要]在免疫反应期间，B细胞归巢于淋巴结（LN ），在那里它们遇到抗原。归巢始于在高内皮小静脉（HEV）的活化内皮细胞上进行捕获和L-选择蛋白依赖性滚动。在识别出存在于HEV上的趋化因子后，B细胞整合素的激活发生了介导的牢固停滞。随后，B细胞爬到外渗点进入LN。外渗可通过腹股沟淋巴结的活体显微镜检查（IVM）进行体内可视化和定量。在这里，我们描述了一个已建立的协议，该协议允许在无菌炎症条件下对LN的B细胞募集的详细体内分析。我们描述了数据采集，出口，定量，并使用专门的软件进行统计分析。LN的IVM是一种强大的技术，可以更好地了解体内炎症过程中B细胞的迁移行为。
[背景]次级LN在战术上定位为通过淋巴系统排出抗原，因此在人体不同部位的免疫反应中起着基本作用。这意味着原位免疫细胞的募集，抗原的遭遇以及免疫反应的增强（Tan and Watanabe，2010; Tavares et ...
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Author:
Date:
2020-11-20
[Abstract] Several studies suggest an important role of lipid metabolism in regulating longevity of Caenorhabditis elegans. Therefore, assays to quantify lipids have enormous value in understanding aging and pathologies associated with it. Approximately 70% of lipid metabolism genes in the nematode have orthologs in humans. Amenability of C. elegans to genetic manipulations has allowed investigations into the role of specific genetic factors in lipid metabolism. Here, we describe a protocol to quantify total triglycerides in C. elegans, which can be extended to studies of the effects of altered environmental and genetic factors on stored fats. This protocol quantifies the picomoles of the triglycerides, in whole worm lysate. Due to the sensitivity of the assay, it could ...
[摘要] [摘要]多项研究表明脂质代谢在调节秀丽隐杆线虫的寿命中具有重要作用。因此，定量脂质的测定在理解衰老和与其相关的病理学方面具有巨大价值。线虫中大约70％的脂质代谢基因在人类中具有直系同源物。秀丽隐杆线虫对基因操作的适应性允许调查特定遗传因素在脂质代谢中的作用。在这里，我们描述了一个协议，以定量总甘油三酯在秀丽隐杆线虫，这CA n可以扩展到环境和遗传因素改变对储存脂肪的影响的研究。该协议量化了整个蠕虫裂解物中甘油三酸酯的皮摩尔。由于测定的敏感性，它可以帮助鉴定总存储脂肪的细微变化，这是显微镜技术无法识别的。
[背景]多余的卡路里通常以三酰基甘油或糖原的形式存储（Cohen，2018年）。甘油三酸酯（TAGs）的异常水平与人类的心脏病，胰腺炎和动脉粥样硬化有关（Cameron等人，1974; Lee等人，2003; Walther和Farese，2012; Onal等人，2017）。TAG被包装到细胞质中的脂滴中。脂质液滴存在于真核细胞中，大小从20 nm到100 µm不等（Stobart等，1986； ...
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