Movie 2.

Confocal z-stack using a 40x/1.25NA glycerol objective with 6x confocal zoom of animals stained with anti-TUBA-2 (green) and 6G10 (magenta) antibodies. Movie progresses from external to internal tissues, showing circular bodywall muscle, then diagonal muscle, then longitudinal muscle. Animal was fixed with Carnoys and primary antibodies detected using fluorophore-conjugated secondary antibodies (TUBA-2 detected with goat anti-rabbit-alexa568 and 6G10 detected with goat anti-mouse-alexa488). Scale bar, 10 microns.