Em ccd camera

For zebrafish, a Leica SP8 (Leica, Bannockburn, IL) laser scanning confocal microscope (LSCM) was used throughout manuscript. A HC PL APO 20x/0.75 IMM CORR CS2 objective, HC PL APO 40x/1.10 W CORR CS2 0.65 water immersion objective, and an HCX Plan Apochromat 63×/1.40–0.06 NA OIL objective were used. Images were acquired using LAS-X software. Images taken with the SP8 LSCM were obtained through lightning, a built-in deconvolution algorithm. A Leica DMi8 (Leica, Bannockburn, IL) with a X-light v2 confocal unit spinning disk was also used, equipped with an 89 North – LDI laser and a Photometrics Prime-95B camera. Optics used were either 10x/0.32 NA air objective, HC PL APO 63X/1.40 NA oil CS2, HC PL APO 40X/1.10 NA WCS2 CORR, a 40X/1.15 N.A. 19 Lamda S LWD, or 100Å~/1.4 N.A. HC Pl Apo oil emersion objective. A Leica M165 FC stereomicroscope equipped with DFC9000 GT sCMOS camera was used for phenotypic analysis of embryos.
For live cell imaging of C. elegans embryos, a spinning disk confocal system was used. The system is equipped with a Nikon Eclipse and is an inverted microscope with a 60X 1.40NA objective, a CSU-22 spinning disc system and a Photometrics EM-CCD camera from Visitech International. Images were obtained every 2 minutes with a 1-micron z-stack step size.

Immobilization of worms for in utero imaging was performed using agarose pads with polystyrene beads as previously described⁷⁰ . RNAi Experiments were performed at 24°C for 16–22 hours. Images were acquired using a spinning disk confocal system with a Nikon Eclipse inverted microscope with a 40 X 1.30 oil objective, a CSU-22 spinning disc system, and a Photometrics EM-CCD camera from Visitech International operated by MetaMorph software (Molecular Devices). Image analysis was performed on Fiji (National Institutes of Health).