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BIFL Data Analyzer (BIFLDA) is intended for sliding analysis of fluorescence decays and correlation functions built from BIFL data (B&H SPC, PicoQuant TimeHarp 200, PicoHarp 300 and Symphotime-64bit) using Maximum Likelihood (ML) method. Inter arrival time distribution (IATD) coincidence analysis is also supported.

Data Management

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  • Import of B&H SPC data (up to four channels)

  • Import of PicoQuant TimeHarp 200, PicoHarp 300 and Symphotime-64bit (up to four channels)

  • Calculation of absolute photon arrival times when possible

  • Macro & micro time processing

  • Automatic and manual Burst selection in the intensity and time lag plot

  • Sliding by either selected bursts or constant time or constant amount of photons

  • Calculation of fluorescence decays and anisotropy

  • Calculation of autocorrelation functions (with and without the normalization) and inter arrival time distribution in linear and quasi-logarithmic time scales

  • Binning of fluorescence decays

  • Calculation and plotting Energy Transfer Efficiency and Fractional Intensity for two color data

  • Calculation and plotting Steady State Polarization and Anisotropy for polarized data

  • Selecting user-defined time regions, start and end analysis channels with cursors

  • Displaying fluorescence and triplet lifetimes histogram

  • Export data to ASCII file

  • Export autocorrelation functions to ConfoCor formatted file

Analysis Methods

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  • Sliding analysis (Maximum Likelihood with multinomial statistics and Levenberg-Marquardt optimization algorithm)

  • Analysis of selected dataset

  • Automatic shift detection

  • Fit with and without convolution

  • Correction for the background

  • Automatically generated initial guesses

  • Parameter fixing and constraints

  • Confidence intervals by standard errors

  • Quality of fit: MLE criterion value and visual inspection of residuals and autocorrelation of residuals

  • FCS analysis

  • IATD coincidence analysis

Analysis Models

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   Fluorescence decay analysis

where:
d is time shift;
b is level of dark noise;
g is background multiplication factor;
с time-uncorrelated background;|
irf(t) is measured instrument response function;
bg(t) is measured background (optional);

Anisotropy analysis

where:
rinf is anisotropy when t → ∞;
rj are pre-exponential factors;
jj are rotational correlation times;
M is number of exponents.
 

Analysis of autocorrelation function for immobilized molecules

where:
cj are pre-exponential factors;
tj are triplet-state lifetimes;
K is number of exponents.
 

Analysis of autocorrelation function for molecules in solution (FCS)

where:
F and t are triplet-state fraction and lifetime;
N is an average number of molecules in observation volume;
Fj are fractions of corresponding diffusion components;
TDj are diffusion times of corresponding diffusion components;
 

Model library is easily extendable