Measurement of Fluorescence Correlation Function by Using Size and Concentration of Fluorescence Particles

Abstract

Measurement of Fluorescence Correlation Function by Using Size and Concentration of Fluorescence ParticlesThe concentration and hydrodynamic radius of nano-sized fluorescence particles diffusing in solution were compared by using fluorescence correlation spectroscopy (FCS), which can measure the variation of the correlation function of a fluorescence signal by size and number of particles. The used nano-sized fluorescence particles are Alex Fluor 647, quantum dots, and fluorescence beads, and three kinds of sample solutions with different concentrations were prepared by dilution to 1/10 and 1/100 with distilled water for each kind of particles. The effective focal volumes were calculated by using the known diffusion coefficient of Alexa Fluor 647 particles, and the diffusion time, number of particles in focal volume, and variation of concentration according to the dilution could be measured by the FCS system. Through this study, we determined that the concentrations of arbitrarily diluted sample solutions can be measured by a home-built FCS setup in the range of 0.1 nM ~ 10 nM and that the diffusion coefficient of the quantum dot was 27±1 µm 2 /s.