Fluorescence-Based Assays for Measuring Doxorubicin in Biological Systems.
Fluorescence-Based Assays for Measuring Doxorubicin in Biological Systems.
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DOI：
10.20455/ros.2016.873
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发表时间：
2016-01-01
期刊：
影响因子：
--
通讯作者：
Li, Y Robert
中科院分区：
文献类型：
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作者：
Kauffman, Melinda K;Kauffman, Megan E;Li, Y Robert
阿霉素在包括体液、细胞和组织在内的生物系统中的检测和测量，有助于了解这种广泛使用的药物在治疗癌症以及引起不良反应方面的作用机制。在这篇文章中，我们首次描述了基于荧光的技术的使用，包括荧光光谱、显微镜和流式细胞术在测量和/或检测生物系统中的阿霉素，包括细胞裂解物和培养的完整细胞。阿霉素的最大激发波长和发射波长分别为470 nm和560 nm。在缓冲液和细胞裂解液中，荧光光谱分析的检测灵敏度小于0.1微米。荧光显微镜显示，无论是绿色荧光还是红色荧光，都能很容易地检测到阿霉素在培养细胞中的浓度依赖积累，但绿色荧光的检测灵敏度更高。流式细胞术也显示阿霉素在细胞悬液中以浓度依赖的方式聚集的敏感检测。上述三种基于荧光的技术简便、灵敏地测量和检测阿霉素，对于研究不同实验条件下阿霉素在癌细胞和正常细胞中的细胞动力学具有重要意义。
Detection and measurement of doxorubicin in biological systems, including body fluids, cells, and tissues, are instrumental in understanding the mechanisms of action of this widely used drug in treating cancer as well as in causing adverse effects. In this article, we, for the first time, characterized the use of fluorescence-based techniques, including fluorescence spectrometry, microscopy, and flow cytometry in measuring and/or detecting doxorubicin in biological systems, including cell lysates and cultured intact cells. We showed that doxorubicin has a maximum excitation and emission wavelength of 470 and 560 nm, respectively. The detection sensitivity by fluorescence spectrometry is less than 0.1 muM in buffers and cell lysates. Fluorescence microscopy demonstrated the readily detection of concentration-dependent accumulation of doxorubicin in cultured cells via either green or red fluorescence, but with green fluorescence showing a higher sensitivity of detection. Flow cytometry also revealed sensitive detection of doxorubicin accumulation in cell suspensions in a concentration-dependent manner. The readily and sensitive measurement and detection of doxorubicin by the above three fluorescence-based techniques has important implications in studying the cellular dynamics of doxorubicin in both cancer and normal cells under various experimental conditions.