Fig. 6: Example of application of HBR-2,5-DM fluorogen in live-cell fluorescence lifetime multiplexing with three 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 three FAST variants expressed simultaneously in live HeLa cells as H2B-P68T, IMS-F62L (mitochondrial intermembrane space), and β4Gal-T1-R52K (Golgi apparatus) fuses. Color-coding represents fluorescence lifetime (τ) or intensity-weighted average lifetime (τi) and is specified on the right. b Result of color assignment to τi ranges specific to R52K, P68T, 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, P68T, or F62L complexes with HBR-2,5-DM, the composite and the individual localizations are shown. Color-coding of τi ranges and phasor clusters are presented in Supplementary Fig. S98. Representative image, similar results in n = 25 cells. Scale bars 10 micrometers.
