Fluorescence detected linear dichroism spectroscopy: A selective and sensitive probe for fluorophores in flow-oriented systems.

Fluorescence detected linear dichroism spectroscopy: A selective and sensitive probe for fluorophores in flow-oriented systems.
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荧光检测线性二色性光谱:流动导向系统中荧光团的选择性和灵敏探针。

DOI:
10.1002/chir.22795
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发表时间:
2018
期刊:
影响因子:
2
通讯作者:
Wemyss AM
Wemyss AM
中科院分区:
化学4区
文献类型:
--
作者:
Wemyss AM

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荧光检测通常增强荧光分析物的灵敏度和选择性。在这项工作中,通过测量流动定向DNA结合配体和M13噬菌体发出的荧光,探索了在线性二色性实验的正常配置中将荧光检测与样品的流动定向相结合的可能性。列出了溴化乙锭、Hoechst 33258和4,6-二脒基-2-苯基吲哚的数据。该技术的理论基础也为在固定直流模式(圆二色性分光偏振仪的正常操作模式)和固定高压电压模式下运行的仪器提供。到达检测器的杂散光的作用,在固定的直流模式下,类似于线性二色性光谱的形状,而不是预期的减少线性二色性的光谱形状的结果,也进行了探讨。
Fluorescence detection typically enhances sensitivity and selectivity for fluorescent analytes. The potential for combining fluorescence detection with flow orientation of the sample in the normal configuration of linear dichroism experiments is explored in this work by measuring the fluorescence emitted from flow‐orientated DNA‐bound ligands and M13 bacteriophage. Data for ethidium bromide, Hoechst 33258, and 4,6‐diamidino‐2‐phenyindole are presented. The theoretical basis of the technique is also presented for instruments running in both the fixed direct‐current mode, which is the normal operation mode of circular dichroism spectropolarimeters, and also in fixed high‐tension voltage mode. The role of the stray light reaching the detector that results in a spectral shape in fixed direct current mode that resembles the shape of a linear dichroism spectrum, rather than the expected reduced linear dichroism, is also explored.
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