for a key.
Software for confocal Multiparameter Fluorescence Imaging and Spectroscopy (MFIS) is further developed and is provided as separate software package which includes AnI-3SF, Margarita and Kristine programs.
As in version 2014, in AnI-3SF each pixel of Multiparameter Fluorescence Image corresponds to “single burst” in solution experiments and all MFD type analysis are applied. Intensity, lifetime, anisotropy, correlation times, correlation amplitudes or any MFD parameter images can be reconstructed [1,2].
New feautures in Ani-3SF are:
- Particle analysis (particle size, particle size distribution, particle center location and surface area measurements) is added to AnI-3SF program since particle sizing and distribution analysis helps in particles aggregation applications.
- In addition, ROI integrated FLIM is possible. Large number of image files could be analyzed as batch task.
- Raster image correlation spectroscopy (RICS) as a new and novel technique for measuring molecular dynamics and concentrations from fluorescence confocal images is also incorporated into AnI-3SF program.
- The intensity images generated from photons number weighted by component FLIM filter values in AnI-3SF program are referred as filtered FLIM images.
- Colocalization analysis used in FRET nanoscope to localize Donor and Acceptor dyes positions on single molecules by STED microscopy using repeated scan of the same image line with donor excitation and then acceptor excitation laser.
Analysis results are saved in formats defined for Margarita program (software used in MFD analysis for 1D and 2D histograms generation and sub-populations selection) and further selection of pixels with similar spectroscopic properties is possible. Selections later are applicable to the images directly in AnI-3SF via Pixel numbers or Selected ROIs.
Kristine program includes few more special functions for fitting RDA distances distributions.