Ixon ultra 897 ccd camera

Adult gravid hermaphrodite worms were dissected in a watch glass filled with Egg Salts medium (118 mM KCl, 3.4 mM MgCl₂, 3.4 mM CaCl₂ and 5 mM HEPES pH 7.4), and embryos were mounted on a fresh 2% agarose pad and covered with an 18 mm×18 mm coverslip (No. 1.5H, Marienfeld). Embryos co-expressing GFP::histone H2B and GFP::γ-tubulin for tracking of centrosomes and nuclei (Fig. 1F–H; Fig. 2C–G; Fig. 3G) were imaged on an Axio Observer microscope (Zeiss) equipped with an Orca Flash 4.0 camera (Hamamatsu), a Colibri.2 light source, and controlled by ZEN software (Zeiss). All other imaging was performed on a Nikon Eclipse Ti microscope coupled to an Andor Revolution XD spinning disk confocal system composed of an iXon Ultra 897 CCD camera (Andor Technology), a solid-state laser combiner (ALC-UVP 350i, Andor Technology), and a CSU-X1 confocal scanner (Yokogawa Electric Corporation), controlled by Andor IQ3 software (Andor Technology). All imaging was performed in temperature-controlled rooms kept at 20°C. Time-lapse sequences were processed and analyzed with Fiji software (Image J version 2.0.0-rc-56/1.51 h).

Mid-log phase cultures of Synechocystis PCC 6803 and Cyanothece PCC 7425 cells harboring the pSB2TΔKm^R-gfp, pSB2T-ccmk1_tsbp1-gfp and pSB2T-maf_S6803-gfp plasmids were placed in sandwiches consisting of two glass coverslips (22 mm diameter, Paul Marienfeld GmbH & Co. KG) one of which being coated with a Poly-L-lysine (Sigma-Aldrich) monolayer. These coverslip sandwiches were sealed and placed inside a home-made sample holder, which was mounted on a Nikon Ti-U inverted microscope coupled with an iXon ULTRA 897 CCD camera (Andor Technology), equipped with a 100x oil immersion (NA 1.45) microscope objective. Epifluorescence images were recorded using an excitation provided by a plasma light source (HPLS245 Thorlabs, Inc.) and an excitation filter (MF469-35 Thorlabs, Inc.), while for super-resolution laser scanning images a 488 nm laser (OBIS, Coherent) was used as an excitation source.
Chlorophyll and GFP fluorescence were recorded using ET655LP (Chroma Technology Corporation) and MF525/39 (Thorlabs, Inc.) emission filters, respectively.