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时间分辨荧光中的仪器响应标准

Instrument response standard in time-resolved fluorescence.

作者信息

Luchowski Rafal, Gryczynski Zygmunt, Sarkar Pabak, Borejdo Julian, Szabelski Mariusz, Kapusta Peter, Gryczynski Ignacy

机构信息

Department of Molecular Biology and Immunology, Center for Commercialization of Fluorescence Technologies, UNTHSC, Fort Worth, Texas 76107, USA.

出版信息

Rev Sci Instrum. 2009 Mar;80(3):033109. doi: 10.1063/1.3095677.

Abstract

The fluorescence of LDS 798 dye in aqueous solution has a very short lifetime of 24 ps, independent of excitation wavelength. The time response of common photon counting detectors depends on the wavelength of the registered photon. In lifetime measurements, the instrument response function (IRF) is usually approximated by the temporal profile of the scattered excitation light. Because lambda(Exc) is typically much shorter than lambda(Em), a systematic error may be present in these measurements. We demonstrate that the fluorescence decay of LDS 798 is a better approximation of IRF, in particular, for avalanche photodiodes used in the near infrared spectral region.

摘要

LDS 798染料在水溶液中的荧光寿命非常短,仅为24皮秒,且与激发波长无关。普通光子计数探测器的时间响应取决于所记录光子的波长。在寿命测量中,仪器响应函数(IRF)通常由散射激发光的时间分布近似表示。由于激发波长(lambda(Exc))通常比发射波长(lambda(Em))短得多,因此这些测量中可能存在系统误差。我们证明,LDS 798的荧光衰减是IRF的更好近似,特别是对于近红外光谱区域中使用的雪崩光电二极管而言。

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