Global analysis in fluorescence correlation spectroscopy and fluorescence lifetime microscopy.
Methods Enzymol
; 518: 145-73, 2013.
Article
em En
| MEDLINE
| ID: mdl-23276539
ABSTRACT
Fluorescence correlation spectroscopy (FCS) and related fluctuation spectroscopy and microscopy methods have become important research tools that enable detailed investigations of the chemical and physical properties of molecules and molecular systems in a variety of complex environments. Information recovery via curve fitting of fluctuation data can present complicating challenges due to limited resolution and/or problems with fitting model verification. We discuss a new approach to data analysis called τFCS that couples multiple modes of signal acquisition, here specifically FCS and fluorescence lifetimes, with global analysis. We demonstrate enhanced resolution using τFCS, including the capability to recover the concentration of both molecular species in a two-component mixture even when the species have identical diffusion coefficients and molecular brightness values, provided their fluorescent lifetimes are distinct. We also demonstrate how τFCS provides useful tools for model discrimination in FCS curve fitting.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Espectrometria de Fluorescência
/
Microscopia de Fluorescência
Idioma:
En
Revista:
Methods Enzymol
Ano de publicação:
2013
Tipo de documento:
Article
País de afiliação:
Estados Unidos