Supplementary Figure 4: Three different structures in U2OS cells labeled with Alexa Fluor 647.
From: Multi-target spectrally resolved fluorescence lifetime imaging microscopy
The three different staining strategies cause small changes in the dye's fluorescence properties, i.e., lifetime (f) and spectrum (g). These changes are sufficient to clearly distinguish between all three structures: (c) Golgi stained with primary rabbit antibody against giantin and Alexa Fluor 647-labeled goat anti-rabbit secondary antibody (magenta), (d) F‑actin stained with Alexa Fluor 647 phalloidin (green), and (e) EdU (nascent DNA) inside the nucleus stained via click chemistry with Alexa Fluor 647 azide (blue). (b) shows the resulting sFLIM image and (a) is the mere intensity image. The brightest pixels belong to 3,396 (a) / 3,396 / 1,204 / 2,553 (c–e) photons. Scale bars, 10 µm.
