SH‐SY5Y were plated on a 35 mm Ibidi Glass Bottom dish at a concentration of 2 × 105 cells per dish. 48 h after seeding, cells were incubated with 1× NucSpot® Live 488 (Biotium Inc., USA) and 1× MitoView® Fix 640 (Biotium Inc., USA) for 30 min at 37°C prior to imaging. TNF was added immediately before starting the image acquisition. Imaging was carried out using an Elyra 7 Lattice‐SIM Super‐Resolution microscope (Zeiss, Oberkochen, Germany) every 2 min for 30 min with a 63× oil objective (Plan‐Apochromat 63×/NA1.4). Image analysis was performed with Imaris 9.9.1 (Bitplane AG, Switzerland) using the Spot (mitochondria) and surface (nucleus) functions. Mitochondrial shortest distance to the nucleus is color‐coded.
Macrophages were plated on at 50,000 cells/well in μ‐Slide 8‐well chambers (80826, Ibidi, Martinsried, Germany) and incubated with 0.5 μM MitoTracker Green (Thermo Fisher Scientific) for 15 min at 37°C prior to imaging. The nuclei were stained using 2 μM Hoechst (Thermo Fisher Scientific). TNF was added immediately before staring video recording. Imaging was carried out using a confocal microscope (LSM800, Carl Zeiss, Oberkochen, Germany) and LD Plan‐Apochromat 40× objective.
Macrophages were plated on at 50,000 cells/well in μ‐Slide 8‐well chambers (80826, Ibidi, Martinsried, Germany) and incubated with 0.5 μM MitoTracker Green (Thermo Fisher Scientific) for 15 min at 37°C prior to imaging. The nuclei were stained using 2 μM Hoechst (Thermo Fisher Scientific). TNF was added immediately before staring video recording. Imaging was carried out using a confocal microscope (LSM800, Carl Zeiss, Oberkochen, Germany) and LD Plan‐Apochromat 40× objective.