Supplementary Figure 2: Five cellular structures of U2OS cells labeled with five fluorophores excitable at 640 nm.
From: Multi-target spectrally resolved fluorescence lifetime imaging microscopy
(a) Mere intensity image. (b) sFLIM composite image consisting of (c) F-actin stained with ATTO 655 phalloidin (green), (d) nucleus stained with DNA intercalating DRAQ5 (blue), (e) nucleoli stained with primary mouse antibody against fibrillarin and Alexa Fluor 633-labeled goat anti-mouse secondary antibody (white), (f) Golgi stained with primary rabbit antibody against giantin and Alexa Fluor 700-labeled goat anti-rabbit secondary antibody (yellow), and (g) microtubules stained with Alexa Fluor 647-labeled rabbit anti-α-tubulin direct antibody (magenta). (h,i) The five corresponding reference patterns: (h) fluorescence decays after the three laser pulses and (i) emission spectra. The brightest pixels belong to 4,139 (a) / 1,895 / 2,159 / 942 / 4,129 / 1,509 (c–g) photons. Scale bars, 10 µm.
