Ff01 605 54

For fluorescence time-lapse microscopy, cells were imaged using a confocal spinning-disk inverted microscope (Nikon TI-E Eclipse) equipped with an Evolve EMCCD camera. The optical sectioning was performed by a Yokogawa motorized confocal head CSUX1-A1. Images were acquired using an illumination system from Roper Scientific (iLasPulsed) with a CFI Plan APO VC oil immersion objective (60X, N.A 1.4) or CFI Plan Fluor oil immersion objective (40X, N.A. 1.3). Z-series were generated using a motorized Z-piezo stage (ASI) by acquiring images with a step size of 0.4 μm. Microscope was controlled with MetaMorph software (Molecular Devices). Temperature, CO₂, and humidity control was performed using a chamlide TC system (TC-A, Quorum technologies). Solid-state 405, 491 and 561 nm lasers (iLas, Roper Scientific) and ET 460/50M (Chroma), ET 525/50M (Chroma), FF01-605/54 (Semrock) emission filters were used for excitation and emission of Hoechst 33342, EGFP and mCherry fluorescence, respectively.

For fluorescence time-lapse microscopy, cells were imaged using a confocal spinning-disk inverted microscope (Nikon TI-E Eclipse) equipped with an Evolve EMCCD camera. The optical sectioning was performed by a Yokogawa motorized confocal head CSUX1-A1. Images were acquired using an illumination system from Roper Scientific (iLasPulsed) with a CFI Plan Fluor oil immersion objective (40X, N.A. 1.3). Z-series were generated every 10 min using a motorized Z-piezo stage (ASI) by acquiring 25 z-plane images with a step size of 1 μm. Microscope was controlled with MetaMorph software (Molecular Devices). Temperature, CO₂, and humidity control was performed using a chamlide TC system (TC-A, Quorum technologies). Solid-state 491 and 561 nm lasers (iLas, Roper Scientific) and ET 525/50M (Chroma) and FF01-605/54 (Semrock) emission filters were used for excitation and emission of EGFP and propidium iodide fluorescence, respectively.