An ImageJ plugin for intensity-based three-filter set (ratiometric) FRET which works with unknown varying donor/accepter ratios, corrects for channel crosstalk and instrument calibration, and yields quantitative FRET E values.
- Download the latest version of Fiji.
- Enable the FRET Imaging update site.
- Once installed, go to Plugins > FRET Imaging > RiFRET.
This software is based on a publication. If you use it in your work, please cite:
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Rebenku, I., Lloyd, C.B., Szöllősi, J. et al. Pixel-by-pixel autofluorescence corrected FRET in fluorescence microscopy improves accuracy for samples with spatially varied autofluorescence to signal ratio. Sci Rep 13, 2934 (2023). https://doi.org/10.1038/s41598-023-30098-w
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Roszik, J., Lisboa, D., Szöllősi, J. and Vereb, G. (2009), Evaluation of intensity-based ratiometric FRET in image cytometry—Approaches and a software solution. Cytometry, 75A: 761-767. https://doi.org/10.1002/cyto.a.20747
If you are using the acceptor photobleaching FRET method, we recommend our other FRET plugin AccPbFRET.