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Dead-time distortion in fluorescence correlation measurements

机译:荧光相关性测量中的死区失真

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摘要

The dead time of the detector significantly distorts the fluorescence correlation function of fluorescent particles in solution. This distortion of the correlation function is similar to the saturation effect of the correlation function in a high-power excitation region. The correlation amplitude is significantly reduced by the dead time. The deviations in the number of molecules and the diffusion time are empirically given by the deviation of the fluorescence intensity linearity. The empirical curves of the deviations can be applied to the systematic error estimation of the parameters. The proportionality of the number of molecules to the concentration of fluorophores is no longer maintained with a large dead time, although almost all of the proportionality of the diffusion time to the inverse diffusion constant remains. This fact makes the dead-time effect different from the saturation effect, which is due to photokinetics. In practice, these distortions can be reduced by use of a smaller excitation power in which the proportionality of the fluorescence intensity is maintained.
机译:检测器的死区时间会严重扭曲溶液中荧光颗粒的荧光相关功能。相关函数的这种失真类似于高功率激励区域中相关函数的饱和效应。停滞时间会大大降低相关幅度。分子数量和扩散时间的偏差由荧光强度线性的偏差根据经验给出。偏差的经验曲线可以应用于参数的系统误差估计。尽管几乎没有扩散时间与逆扩散常数成比例的所有关系,但是在死时间较长的情况下,分子数目与荧光团浓度的成比例关系不再保持。这个事实使空载时间效应不同于饱和效应,这是由于光动力学引起的。实际上,可以通过使用较小的激发功率来减少这些畸变,在该较小的激发功率中,荧光强度的比例得以保持。

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