The QuanTI-FRET method proposes calibrating the instrument and the FRET pair to simply
calculate absolute FRET probabilities from triplet of images acquired under the same conditions
and with the same FRET pair. All the photophysical and instrumental factors are included in this calibration, leaving
the variability of the results to biological origins.
The main contribution of the QuanTI-FRET method has been the single-step
calibration of the two correction factors based on the batching of millions of pixels, thus fully taking
advantage of the statistics of the imaged pixels and making this calibration method extremely robust.
The QuanTI-FRET has been reworked to allow calibration with FRET standards or autocalibration directly for the images of interest! To make quantitative FRET measurement accessible to all laboratories, a software is being prepared as standalone in Python and plug-in for Napari. The Python code is made publicly available as a Git.
"QuanTI-FRET: a framework for quantitative FRET measurements in living cells." A Coullomb, CM Bidan, C Qian, F Wehnekamp, C Oddou, C Albiges-Rizo, DC Lamb et A Dupont. Scientific Reports 10 (1), 1-11 (2020)
"Live-cell quantitative FRET imaging made simple by autocalibration in QuanTI-FRET." J Leblanc, AH Lombard, A Saumureau, S Costrel, J Revilloud, A Coullomb et A Dupont. under review
Co-authors of the code: Alexis Coullomb, Adrien Saumureau and Aurélie Dupont.