Analysis of protein aggregation in sub-femtoliter volume by two-photon-based fluorescence correlation spectroscopy
Journal
Journal of Bionanoscience
Journal Volume
2
Journal Issue
2
Pages
97-101
Date Issued
2008
Author(s)
Abstract
We presented a fluorescence correlation spectroscopy (FCS) based on two-photon microscopic system to identify physical parameters that give rise to fluctuations in fluorescence signal. The excitation volume is a function of the overall instrumental setting, that include the utilized laser power, the laser wavelength, the magnification, and the power of the objectives and other optics for the microscope. For a given excitation volume, the diffusion coefficient of molecules due to Brownian Movement into or out of this excitation volume can be determined. Furthermore, based on the autocorrelation analysis that is dependent on the diffusion coefficients of molecules, the fraction of bound species from measured samples containing the dye labeled anti-rabbit IgG and rabbit IgG can be separated. The mean standard deviation of the diffusion coefficients measured in our two-photon-based system is about 0.064. Moreover, the maximal binding fraction of kinetics mode and end mode between anti-rabbit IgG and rabbit IgG is 60% and 73%, respectively. Copyright © 2008 American Scientific Publishers All rights reserved.
SDGs
Type
journal article