| Quantitative Analysis of Exosome Secretion Rates of Single Cells
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Author:
Date:
2017-02-20
[Abstract] To study the inhomogeneity within a cell population including exosomes properties such as exosome secretion rate of cells and surface markers carried by exosomes, we need to quantify and characterize those exosomes secreted by each individual cell. Here we develop a method to collect and analyze exosomes secreted by an array of single cells using antibody-modified glass slides that are position-registered to each single cell. After each collection, antibody-conjugated quantum dots are used to label exosomes to allow counting and analysis of exosome surface proteins. Detailed studies of exosome properties related to cell behaviors such as responses to drugs and stress at single cell resolution can be found in the publication (Chiu et al., 2016).
[摘要] 为了研究细胞群体内的不均匀性，包括外来体特征，如外来体外分泌细胞分泌速度和外来体携带的表面标志物，我们需要量化和表征每个细胞分泌的那些外来体。在这里，我们开发收集和分析由单个细胞阵列分泌的外来体的方法，使用位置登记到每个单细胞的抗体修饰的玻片。每次收集后，使用抗体缀合的量子点来标记外来体以允许外来体表面蛋白的计数和分析。在出版物（Chiu等人，2016）中可以找到与细胞行为相关的外来物质性质的详细研究，例如对药物的反应和单细胞分辨率的压力。
背景 ...
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| Extraction of Small RNA and qPCR Validation of miRNAs in Vigna mungo
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Author:
Date:
2015-03-05
[Abstract] Small RNAs like microRNAs (miRNAs), small interfering RNAs (siRNAs) and other noncoding RNAs including snRNA and snoRNA have tremendous impact on eukaryotic gene regulation. Extraction of high quality small RNAs is an important prerequisite for experimental analyses of miRNAs. This will prevent RNA degradation and remove associated contaminations including polyphenols, polysaccharides and other secondary metabolites. In this protocol we describe a simple way to isolate small RNAs from the leaf tissues of Vigna mungo combining the protocols of two commercially available kits with some modifications.
[摘要] 小RNA如微小RNA（miRNA），小干扰RNA（siRNA）和其他非编码RNA（包括snRNA和snoRNA）对真核基因调控具有巨大的影响。 高质量小RNA的提取是miRNA的实验分析的重要先决条件。 这将防止RNA降解和去除相关的污染物，包括多酚，多糖和其他次生代谢物。 在这个协议中，我们描述了一种简单的方法，从猕猴桃的叶组织中分离小RNAs，结合两个商业上可用的试剂盒的协议与一些修改。
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