timelapse recordings

praparation of the embryos

• collect embryos on an apple juice plate, 0-3 h
• prepare a coverslide coated with embryo glue (use a pasteur pipette)
• prepare a piece of apple juice agar on a slide
• cover the apple juice plate with 50% Klorix, incubate for 90 s.
• collect the embryos in a net
• wash the plate with water, collect the remaining embryos
• dry the embryos in the net on a paper towel
• transfer the embryos with a fine brush from the net to the piece of agar
• align the embryos (anterior-micropyle to the left, ventral down)
• gently place the coverslide with the the glue on top of the embryos
• flip back the coverslide, the embryos should stick to it
• cover the embryos with 10S voltalef oil (use as little as possible)
• the embryos are ready for microscopy

microscopy

• fix the slide with tape to a holder
• login: user: grosshans, pswd: openlab282
• make sure the the folders pictures and QT movies are empty
• start the programme, open the automation
• choose the objective, insert the polarisation filters above and below the objective,
• select the correct condensor ring, select the correct prism below the objective
• for DIC and bright-field optics, set gain and offset to zero
• make sure that the objective is not in parking position