| 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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| Sodium Current Measurements in HEK293 Cells
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Author:
Date:
2013-08-20
[Abstract] This protocol is used to measure sodium currents from heterologously transfected cells lines, such as HEK293 cells. Standard whole cell patch clamp technique is used to assess ion channel function. This protocol has been used to study cardiac type Nav1.5 sodium channel, and in particular to compare wild-type and mutant channels related to Brugada Syndrome (BrS). Mutations related to BrS provoke a loss of function of the cardiac sodium channel. This is evidenced by a reduction of sodium current density and/or alterations of the activation or inactivation kinetic parameters, such as a slow recovery from inactivation. These effects could explain the alteration of the cardiac action potential that leads to the characteristic ST elevation observed in the electrocardiogram of the ...
[摘要] 该方案用于测量异源转染的细胞系如HEK293细胞的钠电流。标准全细胞膜片钳技术用于评估离子通道功能。该方案已经用于研究心脏型Na v 1.5钠通道，特别是用于比较与Brugada综合征（BrS）相关的野生型和突变型通道。与BrS相关的突变引起心脏钠通道功能的丧失。这通过钠电流密度的降低和/或活化或失活动力学参数的改变，例如从失活缓慢恢复来证明。这些效应可以解释导致在Brugada综合征患者的心电图中观察到的特征性ST升高的心脏动作电位的改变。该方案具有一些变化，适合于研究与心脏类型钠通道功能的改变相关的其它心律失常以及用于研究其它电压依赖性钠通道。
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