Fig. 5: Example of application of HBR-2,5-DM fluorogen in live-cell fluorescence lifetime multiplexing with two FAST variants.
From: Fluorescence lifetime multiplexing with fluorogen activating protein FAST variants
a Total intensity and color-coded FLIM images with mono- (τ) or biexponential (τi) fit of HBR-2,5-DM in complexes with two FAST variants expressed simultaneously in live HeLa cells as H2B-F62L and vimentin-R52K fusions. Color-coding represents fluorescence lifetime (τ) or intensity-weighted average lifetime (τi) and is specified on the right. b Result of color assignment to τ ranges specific to R52K or F62L complexes with HBR-2,5-DM, the composite and the individual localizations are shown. c Result of color assignment to phasor clusters circles specific to R52K or F62L complexes with HBR-2,5-DM, the composite and the individual localizations are shown. Representative image, similar results in n = 18 cells. More data and other examples can be found in the supporting information (Supplementary Figs. S80–96). Scale bars 10 micrometers.
