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利用基于流动的微流控装置中的生物发光快速定量活细胞受体。

Rapid quantification of live cell receptors using bioluminescence in a flow-based microfluidic device.

机构信息

Department of Biomedical Engineering, 9 Engineering Drive 1, Block EA, #03-12, National University of Singapore, Singapore, 117575.

出版信息

Small. 2015 Feb 25;11(8):943-51. doi: 10.1002/smll.201401674. Epub 2014 Oct 21.

Abstract

The number of receptors expressed by cells plays an important role in controlling cell signaling events, thus determining its behaviour, state and fate. Current methods of quantifying receptors on cells are either laborious or do not maintain the cells in their native form. Here, a method integrating highly sensitive bioluminescence, high precision microfluidics and small footprint of lensfree optics is developed to quantify cell surface receptors. This method is safe to use, less laborious, and faster than the conventional radiolabelling and near field scanning methods. It is also more sensitive than fluorescence based assays and is ideal for high throughput screening. In quantifying β(1) adrenergic receptors expressed on the surface of H9c2 cardiomyocytes, this method yields receptor numbers from 3.12 × 10(5) to 9.36 × 10(5) receptors/cell which are comparable with current methods. This can serve as a very good platform for rapid quantification of receptor numbers in ligand/drug binding and receptor characterization studies, which is an important part of pharmaceutical and biological research.

摘要

细胞表达的受体数量在控制细胞信号事件中起着重要作用,从而决定其行为、状态和命运。目前用于定量细胞受体的方法要么繁琐,要么不能保持细胞的天然形态。在这里,开发了一种将高灵敏度的生物发光、高精度的微流控和无透镜光学的小足迹结合起来的方法,用于定量细胞表面受体。这种方法使用安全、劳动强度低、速度比传统的放射性标记和近场扫描方法快。它也比基于荧光的测定法更灵敏,是高通量筛选的理想选择。在定量 H9c2 心肌细胞表面表达的 β(1)肾上腺素能受体时,该方法得到的受体数量为 3.12×10(5)至 9.36×10(5)受体/细胞,与目前的方法相当。这可以作为一个很好的平台,用于快速定量配体/药物结合和受体特性研究中的受体数量,这是药物和生物学研究的重要组成部分。

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