1kx1k emccd camera

3-amino-benzoic acid ester (Tricaine) was used to anesthetize
animals for imaging. After anesthetization, they were covered in 0.8%
low-melting point agarose in glass-bottomed 35 mm Petri dishes and mounted
on their right side. Images were captured using a spinning disk confocal
microscope custom build by 3i technology© with: Zeiss Axio Observer
Z1 Advanced Mariana Microscope, X-cite 120LED White Light LED System, filter
cubes for GFP and mRFP, a motorized X,Y stage, piezo Z stage, 20X Air (0.50
NA), 63X (1.15NA), 40X (1.1NA) objectives, CSU-W1 T2 Spinning Disk Confocal
Head (50 uM) with 1X camera adaptor, and an iXon3 1Kx1K EMCCD camera,
dichroic mirrors for 446, 515, 561, 405, 488, 561, 640 excitation, laser
stack with 405 nm, 445 nm, 488 nm, 561 nm and 637 nm with laser stack
FiberSwitcher, photomanipulation from vector© high speed point
scanner ablations at diffraction limited capacity, Ablate!TMª
Photoablation System (532 nm pulsed laser, pulse energy 60J @ 200 HZ).
Time-lapse images were collected every 5 min for 24 hr. Images were
processed using Slidebook software and ImageJ. Only brightness and contrast
were adjusted for all images.

All embryos were dechorionated at 48 hpf and anesthetized with 3-amino-benzoic acid ester. Anesthetized embryos were mounted in 0.8% low-melting point agarose and mounted on their right side in 35 mm Petri dishes with glass bottoms. For imaging, a spinning disk confocal microscope from 3i technology© was used. It is equipped with a Zeiss Axio Observer Z1 Advanced Mariana Microscope with X-cite 120LED White Light LED System and filter cubes for GFP and mRFP, a motorized X,Y stage, piezo Z stage, 20X Air (0.50 NA) objective with 2 mm working distance, 63X (1.15NA) water objective with 0.66 mm working distance, 40X (1.1NA) water objective with 0.62 mm working distance, CSU-W1 T2 Spinning Disk Confocal Head (50 uM) with 1X camera adapter, andor iXon3 1Kx1K EMCCD camera, dichroic mirrors for 446, 515, 561, 405, 488, 561, 640 excitation, laser stack containing 405 nm, 445 nm, 488 nm, 561 nm and 637 nm with laserstack FiberSwitcher that has 250 uS switch time, photomanipulation with vector© high speed point scanner ablations at diffraction limited capacity, Ablate!TM© Photoablation System (532 nm pulsed laser, pulse energy 60 J @ 200 HZ). Time lapse images were taken every 5 min over 24 h. Adobe Illustrator and ImageJ were used to process the images and enhance the brightness and contrast.