Fig. 4: flimGANE enables accurate FRET characterization from low-photon-count data.

a Schematic of CFP-g-YFP FRET pair interacting with glucose and normalized excitation and emission spectra of CFP and YFP. b Intensity contrast and FLIM images of CFP generated by TD_LSE, TD_MLE, DFD_LSE and flimGANE before and after adding 2 mM glucose. c Energy transfer efficiency, E, versus glucose concentration (error bars, standard deviation errors on the parameter estimate, n = 1507~6824 pixels). The flimGANE FRET data can be well fitted by a sigmoidal curve (R2 = 0.92).