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荧光相关谱测量技术研究
Studies on a New Technique of Fluorescence Correlation Spectroscopy
【摘要】 荧光相关谱(fluorescence correlation spectroscopy,FCS)是对处于热平衡态条件下的荧光分子发出的荧光强度涨落进行时间相关处理的一种单分子检测方法,能够直接测量分子在溶液里的扩散系数和浓度。影响FCS测量扩散系数准确性的因素有分子量子效率,测量时间,样本折射率和温度偏差等。用FCS分别测量溶有荧光染料罗丹明6G(rhodamine 6G,Rh.6G)和青色素Cy5甘油水溶液的粘滞系数,实验结果表明:荧光分子的量子效率是影响测量准确性的重要因素,要求其每秒发射的光子数目(photon counts per second,cps)至少达到1 000(photons/s)。
【Abstract】 Fluorescence correlation spectroscopy(FCS),a kind of single molecule technique,performs temporally correlative analysis of spontaneous intensity fluctuation of fluorescence from thermal equilibrium with direct measurements of the diffusion coefficient and the concentration of fluorescence molecules in aqueous solution.Some factors,such as the quantum coefficient,the measurement time,the refractive index of the sample and the temperature deviation,make effects on the measurement accuracy.In the thesis,FCS is used to measure the viscosity of the glycerin-water solution with Rhodamine 6G and Cy5, respectively and the experiment results show that the quantum coefficient plays an important role in the measurement accuracy with 1 000 photons/sec as the minimal level.
【Key words】 fluorescence correlation spectroscopy(FCS); single molecule technique; viscosity; the quantum coefficient; the refractive index;
- 【文献出处】 激光生物学报 ,Acta Laser Biology Sinica , 编辑部邮箱 ,2007年02期
- 【分类号】Q6-33
- 【被引频次】3
- 【下载频次】350