Wemyss Alan M, Chmel Nikola P, Lobo Daniela P, Sutherland John A, Dafforn Timothy R, Rodger Alison
Department of Chemistry and MOAC Doctoral Training Centre, University of Warwick, Coventry, UK.
Department of Chemistry and Physics, Augusta University, Augusta, Georgia, USA.
Chirality. 2018 Mar;30(3):227-237. doi: 10.1002/chir.22795. Epub 2018 Jan 3.
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.
荧光检测通常会提高对荧光分析物的灵敏度和选择性。在这项工作中，通过测量流动取向的与DNA结合的配体和M13噬菌体发出的荧光，探索了在正常线性二色性实验配置中结合荧光检测与样品流动取向的可能性。给出了溴化乙锭、Hoechst 33258和4,6-二脒基-2-苯基吲哚的数据。还给出了该技术在固定直流模式（这是圆二色性分光偏振计的正常操作模式）以及固定高压模式下运行的仪器的理论基础。还探讨了到达探测器的杂散光的作用，这种杂散光在固定直流模式下导致光谱形状类似于线性二色性光谱的形状，而不是预期的降低的线性二色性。
