title: Vectorized data acquisition and fast triple-correlation integrals for Fluorescence Triple Correlation Spectroscopy.
abstract: Fluorescence Correlation Spectroscopy (FCS) is widely used to quantitate reaction rates and concentrations of moleculesin vitroandin vivo. We recently reported Fluorescence Triple Correlation Spectroscopy (F3CS), which correlates three signals together instead of two. F3CS can analyze the stoichiometries of complex mixtures and detect irreversible processes by identifying time-reversal asymmetries. Here we report the computational developments that were required for the realization of F3CS and present the results as the Triple Correlation Toolbox suite of programs. Triple Correlation Toolbox is a complete data analysis pipeline capable of acquiring, correlating and fitting large data sets. Each segment of the pipeline handles error estimates for accurate error-weighted global fitting. Data acquisition was accelerated with a combination of off-the-shelf counter-timer chips and vectorized operations on 128-bit registers. This allows desktop computers with inexpensive data acquisition cards to acquire hours of multiple-channel data with sub-microsecond time resolution. Off-line correlation integrals were implemented as a two delay time multiple-tau scheme that scales efficiently with multiple processors and provides an unprecedented view of linked dynamics. Global fitting routines are provided to fit FCS and F3CS data to models containing up to ten species. Triple Correlation Toolbox is a complete package that enables F3CS to be performed on existing microscopes.
authors.affiliation: Dept. of Molecular Biology, The Scripps Research Institute, 10550 N. Torrey Pines Rd., La Jolla CA 92037, USA ; Dept. of Chemistry, The Scripps Research Institute, 10550 N. Torrey Pines Rd., La Jolla CA 92037, USA ; The Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 N. Torrey Pines Rd., La Jolla CA 92037, USA.
authors.forename: William K
authors.initials: WK
authors.lastname: Ridgeway
authors.forename: David P
authors.initials: DP
authors.lastname: Millar
authors.forename: James R
authors.initials: JR
authors.lastname: Williamson
keywords: Data Acquisition; F3CS; FCS; Fluorescence; Fluorescence Triple Correlation Spectroscopy; Multiple-tau