Csu 22 unit

Amniotic cells, glass-bottom 35 mm u-dishes (Ibidi GmbH, Germany) coated with fibronectin, were co-transfected with H2B-GFP and pmRFP-tubulin expression plasmids (Addgene, MA, USA) using Lipofectamine 3000 transfection reagent and according to the manufacturer's instructions. Live cell imaging was performed 48 hr following transfection under a spinning-disk confocal system Andor Revolution XD (Andor Technology, UK) coupled to an Olympus IX81 inverted microscope (Olympus, UK) equipped with an electron-multiplying CCD iXonEM Camera and a Yokogawa CSU-22 unit based on an Olympus IX81 inverted microscope. Two laser lines at 488 and 561 nm were used for the excitation of GFP and pmRFP and the system was driven by IQ software (Andor Technology, UK). Z-stacks (0.8–1.0 μm) covering the entire volume of the mitotic cells were collected every 1.5 min with a PLANAPO 60×/1.4 NA objective. ImageJ was used to process all the videos.

Four-dimensional data sets were collected with Andor Revolution XD spinning-disk confocal system (Andor Technology, Belfast, UK), equipped with an electron-multiplying charge-coupled device iXonEM Camera and a Yokogawa CSU 22 unit based on an Olympus IX81 inverted microscope (Olympus, Southend-on-Sea, UK). Two laser lines at 488 and 561 nm were used for the excitation of GFP and mCherry and the system was driven by Andor IQ software. Z-stacks (0.8–1.0 μm) covering the entire volume of the mitotic cells were collected every 1.5 min with a PlanApo ×60/1.4 NA objective. All images represent maximum-intensity projections of all z-planes. ImageJ/Fiji software was used to edit the movies.

Fibroblasts were grown in ibiTreat polymer‐coated 35‐mm μ‐dishes (Ibidi GmbH, Germany) and imaged using the Andor Revolution XD spinning‐disk confocal system (Andor Technology, Belfast, UK), equipped with an electron‐multiplying charge‐coupled device iXonEM Camera and a Yokogawa CSU 22 unit based on an Olympus IX81 inverted microscope (Olympus, Southend‐on‐Sea, UK). The system was driven by Andor IQ software, and laser lines at 488 and 561 nm were used for excitation of GFP and mCherry, respectively. Z‐stacks (0.8–1.0 μm) covering the entire volume of individual mitotic cells were collected every 1.5 min using a PlanApo 60×/1.4 NA objective. ImageJ/Fiji software was used to edit the movies in which every image represents a maximum intensity projection of all z‐planes.