Probability distribution analysis (PDA) method for the analysis of fluorescence resonance energy transfer (FRET) signals to determine with high precision the originating value of a shot-noise-limited signal distribution. PDA theoretical distributions are calculated explicitly including crosstalk, stochastic variations, and background and represent the minimum width that a FRET distribution must have. In this way an unambiguous distinction is made between shot-noise distributions and distributions broadened by heterogeneities.
Simultaneously and effectively extracts highly resolved information from FRET distributions.
Gaussian distribution of Donor-Acceptor distances is modeled in the PDA method.
Corresponding fit routines for single state, N states, N Gaussian distributed states, Model Free distributed states (Maximum of Entropy) and two states with dynamics are realized.
Rate constants can be recovered.
Steady state anisotropy is defined with very high precision from single molecule data.
