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Software | Fluorescence Correlation Spectroscopy

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Package FCS (570 Mbyte), Installation notes

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Kristine software-correlator and fitting program yields full correlation curves covering more than 12 orders of magnitude in time [1]. An advanced time-correlated single photon counting (TCSPC) technique simultaneously provides traditional autocorrelation fluorescence correlation (FCS) or cross correlation (FCCS) from selected molecules (Burst IDs) with and without color specific TAC gates which makes analysis more powerful.

Software gives opportunity for all potential applications of technique using pulsed or cw laser excitation: antibunching effects, rotational diffusion and fluorescence resonance energy transfer (FRET) as are discussed.

The combination of FCS and fluorescence lifetime (FLCS) [2] is implemented into software correlator. Utilizing advantage of MFD data (color, time and polarization resolved data) more FRET state sensitive filters from stacked 4 decay histograms of green parallel-green perpendicular-red parallel-red perpendicular is possible to generate and curves of different FRET species cross correlation could be calculated. We refer to this technique as filtered FCS later [3].

Direct generation of necessary theoretical filters for lifetime, polarization or lifetime and polarization (anisotropy) dependent signals is possible based on build into software fluorescence decay generator (for single color only).

Kristine software contains around 116 (defined with up to 32 parameters and 32 constants) fitting functions.

Global fit of multiple correlation curves with different fitting functions for each curve is possible.

 References

 [1] Felekyan S., et al., Rev. Sci. Instr. 76, 083104 (2005)

 [3] Felekyan S., et al., ChemPhysChem. 13, 1036-1053 (2012)

 [2] Böhmer M., et al., Chem. Phys. Lett. 353, 439 (2002)

 
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