ac702474u_si_002.pdf (1.16 MB)
Spectrally Resolved Fluorescence Correlation Spectroscopy Based on Global Analysis
journal contribution
posted on 2008-05-01, 00:00 authored by Michael J. R. Previte, Serge Pelet, Ki Hean Kim, Christoph Buehler, Peter T. C. SoMulticolor fluorescence correlation spectroscopy has been
recently developed to study chemical interactions of
multiple chemical species labeled with spectrally distinct
fluorophores. In the presence of spectral overlap, there
exists a lower detectability limit for reaction products with
multicolor fluorophores. In addition, the ability to separate bound product from reactants allows thermodynamic
properties such as dissociation constants to be measured
for chemical reactions. In this report, we utilize a spectrally resolved two-photon microscope with single-photon
counting sensitivity to acquire spectral and temporal
information from multiple chemical species. Further, we
have developed a global fitting analysis algorithm that
simultaneously analyzes all distinct auto- and cross-correlation functions from 15 independent spectral channels. We have demonstrated that the global analysis
approach allows the concentration and diffusion coefficients of fluorescent particles to be resolved despite the
presence of overlapping emission spectra.