Global analysis of fluorescence fluctuation data.
Over the last decade the number of applications of fluorescence correlation spectroscopy (FCS) has grown rapidly. Here we describe the development and application of a software package, FCS Data Processor, to analyse the acquired correlation curves. The algorithms combine strong analytical power with flexibility in use. It is possible to generate initial guesses, link and constrain fit parameters to improve the accuracy and speed of analysis. A global analysis approach, which is most effective in analysing autocorrelation curves determined from fluorescence fluctuations of complex biophysical systems, can also be implemented. The software contains a library of frequently used models that can be easily extended to include user-defined models. The use of the software is illustrated by analysis of different experimental fluorescence fluctuation data sets obtained with Rhodamine Green in aqueous solution and enhanced green fluorescent protein in vitro and in vivo.
Skakun
Victor V
VV
Department of Systems Analysis, Belarusian State University, Minsk 220050, Belarus.
Hink
Mark A
MA
Digris
Anatoli V
AV
Engel
Ruchira
R
Novikov
Eugene G
EG
Apanasovich
Vladimir V
VV
Visser
Antonie J W G
AJ
D000465:Algorithms; D000818:Animals; D001703:Biophysics / Q000379:methods*; D002138:Calibration; D004023:Dictyostelium / Q000378:metabolism; D004058:Diffusion; D005456:Fluorescent Dyes / Q000494:pharmacology; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D015233:Models, Statistical; D008962:Models, Theoretical; D010710:Phosphates / Q000737:chemistry; D012235:Rhodamines / Q000494:pharmacology; D012984:Software; D013223:Statistics as Topic / Q000379:methods*; D013997:Time Factors
D005456:Fluorescent Dyes; D010710:Phosphates; D012235:Rhodamines; D049452:Green Fluorescent Proteins