Quantifying Cellular Internalization with a Fluorescent Click Sensor,ACS Sensors

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Quantifying Cellular Internalization with a Fluorescent Click Sensor
ACS Sensors ( IF 8.2 ) Pub Date : 2018-04-20 00:00:00 , DOI: 10.1021/acssensors.8b00219
Laura I. Selby _{1,

2} , Luigi Aurelio ₁ , Daniel Yuen ₁ , Bim Graham ₁ , Angus P. R. Johnston _{1,

2}

Affiliation

The ability to determine the amount of material endocytosed by a cell is important for our understanding of cell biology and in the design of effective carriers for drug delivery. To quantify internalization by fluorescence, the signal from material remaining on the cell surface must be differentiated from endocytosed material. Sensors for internalization offer advantages over traditional methods for achieving this as they exhibit improved sensitivity, allow for multiple fluorescent markers to be used simultaneously, and are amenable to high-throughput analysis. We have developed a small fluorescent internalization sensor, similar in size to a standard fluorescent dye, that can be conjugated to proteins and uses the rapid and highly specific bio-orthogonal reaction between a tetrazine and a trans-cyclooctene group to switch off the surface signal. The sensor can be attached to a variety of materials using simple chemistry and is compatible with flow cytometry and fluorescence microscopy, making it a useful tool to study the uptake of material into cells.

中文翻译：

用荧光点击传感器定量细胞内在化

确定细胞内吞物质的数量的能力对于我们对细胞生物学的理解以及在设计有效的药物递送载体中很重要。为了通过荧光定量内在化，必须将细胞表面残留物质的信号与内吞物质区分开来。与传统方法相比，用于内部化的传感器具有优势，因为它们显示出更高的灵敏度，允许同时使用多个荧光标记并且适合进行高通量分析。我们开发了一种小型荧光内在化传感器，大小与标准荧光染料相似，可以与蛋白质结合，并使用四嗪与反式之间的快速且高度特异性的生物正交反应-环辛烯基团可关闭表面信号。该传感器可以使用简单的化学方法连接到多种材料，并且与流式细胞仪和荧光显微镜兼容，使其成为研究材料向细胞吸收的有用工具。

更新日期：2018-04-20

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