Measurement of the Diffusion Coefficients of Fluorescence Beads and Quantum Dots by Using Fluorescence Correlation Spectroscopy

FCS (fluorescence correlation spectroscopy) is a technique used to determine the dynamic characteristics of particles in solution and became common in the field of biophysics with the development of the confocal microscope, the high-speed photo-detector, and real-time data acquisition systems. In this study, the FCS system was composed of a commercial fluorescence microscope, a He-Ne laser (632.8 nm), a data. acquisition board, and a software correlator written with LabVIEW. Autocorrelation functions were obtained using the measured fluorescence fluctuations of fluorescent beads and Q-dots (quantum dots) coated with carboxylate in distilled water. The diffusion coefficients of the beads and the Q-dots in distilled water and PBS (phosphate buffered saline) solution were obtained using the viscosity of water and the bead size. Also, using the bead size, we calculated the viscosity of the PBS solution, and we compared the Q-dots in water and in the PBS solution. The result showed that the viscosity of the PBS solution was 2.5 times greater than that of water, and that the sizes of Q-dots in water and PBS solution were one-third and one-sixth smaller than the known values, respectively, due to the pH variation in the solutions.