Eclipse ti u inverted microscope

Gravid hermaphrodites were dissected in osmotically balanced blastomere culture medium (Shelton and Bowerman, 1996 (link)) and the extracted embryos mounted on a 2% agarose pad. Dual fluorescence and DIC time lapse microscopy was performed at room temperature with a 60x CFI Plan Apochromat Lambda (NA 1.4) objective on a Nikon Eclipse Ti-U Inverted Microscope connected to an Andor Zyla 4.2 sCMOS camera, or with a 63x Plan-Apochromat (NA 1.4) objective on a Zeiss ObserverD.1 inverted microscope connected to the same type of camera. One frame was captured every 10 s, and a z-stack was acquired at every time point, covering 20 µm, with a distance of 0.7 µm between focal planes. Time lapse images in Figure 5A,C,J,K; Figure 3—figure supplement 1E; Figure 5—figure supplement 1B were acquired at 23°C using an inverted Olympus IX 81 microscope equipped with a Yokogawa spinning disk CSU - W1 with a 63x (NA 1.42 U PLAN S APO) objective and a 16-bit PCO Edge sCMOS camera. Images were obtained using a 488-nm solid-state laser at 60% laser power, with an exposure time of 400 ms. Embryos were imaged every 5 s, and a z-stack was acquired at every time point, covering 2 µm of the embryo cortex, with a distance of 0.5 µm between focal planes (i.e. four cortical planes).