Ec plan neofluar 40x na1

Fixed immunostained samples were imaged on a Zeiss LSM880 laser scanning confocal microscope. Embryos were mounted in microdroplets of 5μg/ml Hoechst 33342 in PBS on glass-bottomed dished (Maktek) coated with mineral oil (Sigma). Embryos were imaged along the entire z-axis with 1μm step using an oil-immersion Zeiss EC Plan-Neofluar 40x/NA1.3 with a 0.17mm working distance. Laser power was measured prior to each imaging session and adjusted to maintain consistent power output across experiments to reduce technical variability.

Fixed immunostained samples were imaged on a Zeiss LSM880 laser scanning confocal microscope. Pre-implantation embryos were mounted in microdroplets of 5μg/ml Hoechst 33342 in PBS+/+ on glass-bottomed dished (MatTek) coated with mineral oil (Sigma). Embryos were imaged along the entire z-axis with 1μm step using an oil-immersion Zeiss EC Plan-Neofluar 40x/NA 1.3 with a 0.17mm working distance. For post-implantation embryos, a similar setup was used but with an air Plan-Apochromat 20x/NA 0.75 objective.
Low magnification pooled embryo images and E9.5 full embryos were imaged on a Zeiss Axio Zoom.Vie using a PlanNeoFluar Z 1x/0.25 FWD 56mm objective. Pseudo-stacking (Zeiss ZEN Blue) was used for E9.5 images.