Fig. 4
From: Resonant out-of-phase fluorescence microscopy and remote imaging overcome spectral limitations
Speed OPIOM selectively discriminates RSFP signals in the presence of interfering RSFPs. Tuning the illumination parameters enables the selective imaging of RSFPs exhibiting similar resonance conditions but opposite photochromism (a) or distinct resonance conditions using light modulation, either sequentially at one frequency (e) or simultaneously at multiple frequencies (i). Pre-OPIOM and speed OPIOM images tuned to the resonance of Dronpa-3 and Kohinoor (b–d), Dronpa-2 (f, g) and Dronpa (h), or tuned to optimize orthogonality between Dronpa-2 and Padron (j–l). Systems: Fixed HeLa cells expressing βGal-Dronpa-3 and H2B-Kohinoor (b–d), H2B-Dronpa-2 and Lyn11-Dronpa (f–h), or H2B-Dronpa-2 and Mito-Padron (j–l). Localizations: βGal (cytoplasm), H2B (nucleus), Lyn11 (cell membrane). The images were recorded at 37 °C. Scale bars: 20 µm. See Supplementary Tables 1 and 2 for the acquisition conditions
