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The new advanced FAST (Fluorescence Analysis Software Technology) package for the analysis of fluorescence kinetics is the result of a collaboration between Edinburgh Instruments and Alango.
FAST (Fluorescence Analysis Software Technology) performs advanced analysis of Fluorescence Kinetics. It effectively combines a user friendly, graphical interface with sophisticated mathematical algorithms for solving the associated ill-posed optimization problems.
FAST software is based on Alango proprietary and well proven computational algorithms providing fast and reliable automatic fitting. The two main proprietary algorithms used are: Global Least Squares Analysis (GLSA) minimization algorithm and Non-negative Truncated Singular Value Decomposition (NTSVD) approach.
GLSA minimization algorithm and software library were developed for the
treatment of unstable, non-linear least squares optimisation problems with
multiple local minima. Performance of traditional Marquardt-Levenberg algorithm
is generally inadequate for these types of problems that are typical for
multiset/multimodel fitting tasks.
Exceptional robustness of GLSA approach is achieved by:
|-||Ensuring that only reliable directions are considered on each iteration step. Singular value decomposition of the corresponding Jacobian matrix provides the necessary information|
|-||Adaptive step length strategy used for numerical differentiation while computing the Jacobian matrix|
|-||Singular Value Decomposition - the most effective way for stabilization of ill-posed mathematical problems|
|-||Non Negative Least Squares (NNLS) algorithm|
The Global Analysis routine implements a proprietary algorithm for reliable non-linear least squares data analysis with minimal dependence on initial conditions and minimal probability of resulting at a local minimum. This is the only commercial, general-purpose library that allows the simultaneous fitting of several data sets while some of optimized parameters are linked together. The classical, one data set, one model function fit is implemented as a particular case of a more general multi-model, multi-set problem. This functionality is achieved by using GLSA software library.
Discrete Components Analysis provides robust multi-exponential analysis (up to four components) with automatic choice of initial conditions. Robust convergence in ensured by using GLSA algorithm. Shift and background are estimated automatically or fixed to predefined values. The fitting channel range is easily changed if the full range is inadequate.
Robust, easy to use Distribution Analysis allows recovering of lifetime distribution function with up to 200 logarithmically or linear scaled distribution columns and simultaneous (optional) estimation of shift and background parameters. The functionality is based on NTSVD software library. The fitting channels range is easily changed. The optimal level or "regularization" is set automatically. A priori information about the distribution may be utilized by using a "resolution slider" that allows making other than default trade-off between the smoothness and fitting quality.
Six different anisotropy functions can be selected:
Anisotropy analysis is based on GLSA library providing simultaneous fitting of parallel and crossed polarization data with automatic estimation of G factor.
These functionalities will be added in the upcoming updates. Updates will be available for free download from our web site.
FAST includes many other new features that make it unique software for fluorescence kinetics analysis:
Besides simple ASCII formats, FAST supports native data formats of the following manufacturers: