The software also provides for the convenient display of histograms of each fitted parameter, as well as correlation and error bar plots.
This allows particular features to be visually highlighted and structural, as well as lifetime, information to be represented in a single image. The fitting client enables users to generate false-colour maps of their selected model parameters, to adjust the range over which the colour map is applied for each parameter, and to produce a ‘merged’ image where the intensity of the false-colour, at each pixel, is modulated by the relative brightness of the original data. FLIMfit can also be applied to time-resolved fluorescence anisotropy data and in general to complex models for multichannel FLIM data. Global fitting can be applied across an arbitrary number of images or regions of interest, including across time series and/or arrays of fluorescence lifetime images.
to analyse FRET data with complex donor decay profiles). For global fitting the model parameters can be linked if required (e.g. FLIMfit can also provide global fitting (where the lifetime components are assumed to be spatially invariant across images or regions of interest and are fitted simultaneously by minimising a global χ² ). to define regions of interest) and can be used in conjunction with CellProfiler.įLIM data can be analysed on a pixelwise basis or utilising global binning (where all the pixels in a region of interest are combined to create a “single pixel” with more photons). Fitting FLIM dataįLIMfit is able to fit lifetime data to monoexponential decay profiles or to complex decay profiles with multiple exponential decay components. FLIM data previously imported to an rver is also supported, as described in Supported File Formats. FLIMfit now supports OME-TIFF and most types of FLIM files using the Bio-Formats library. FLIM data can also be imported directly from disk. Users can import raw FLIM data using OMERO, fit it to a fluorescence decay model using the FLIMfit OMERO client and then save the resultant images to an rver or to disk.
Please see Warren et al, 2013 for further details.ĭownload FLIMfit 5.1.1 for Windows or Mac This makes it useful to analyse time series or multiwell plate FLIM data. It can incorporate image segmentation and utilises a highly efficient algorithm that can globally fit FLIM data with modest photon numbers to complex decay models, across hundreds of fields of view, requiring only tens of seconds of processing time on a reasonable desktop computer. FLIMfit is a software tool and an OMERO client that is designed to facilitate analysis and visualisation of time-resolved data from FLIM (Fluorescence Lifetime Imaging) measurements including time-correlated single photon counting (TCSPC) and wide-field time-gated imaging.