Animals were either anaesthetized with 0.2 mg/ml MS-222 (PHARMAQ, UK),
or (in case of GCaMP imaging) immobilized with 0.5 mg/ml of the non-depolarizing
neuromuscular junction blocker mivacurium chloride (Abcam). For confocal
microscopy, animals were mounted laterally in 1% ultrapure low melting point
agarose (Invitrogen) on a glass coverslip. The coverslip was flipped over on a
glass slide with a ring of high-vacuum grease filled with a drop of
Danieau’s solution to prevent drying out of the agarose. For lightsheet
microscopy, embryos were mounted upright in low melting point agarose in a
U-shaped glass capillary (Leica). After imaging, the animals were either
sacrificed or released from the agarose using microsurgery blades and kept
individually until further use.
or (in case of GCaMP imaging) immobilized with 0.5 mg/ml of the non-depolarizing
neuromuscular junction blocker mivacurium chloride (Abcam). For confocal
microscopy, animals were mounted laterally in 1% ultrapure low melting point
agarose (Invitrogen) on a glass coverslip. The coverslip was flipped over on a
glass slide with a ring of high-vacuum grease filled with a drop of
Danieau’s solution to prevent drying out of the agarose. For lightsheet
microscopy, embryos were mounted upright in low melting point agarose in a
U-shaped glass capillary (Leica). After imaging, the animals were either
sacrificed or released from the agarose using microsurgery blades and kept
individually until further use.