1.4 na hcx pl apochromat oil objective

Embryos were raised at 25°C, collected, dechorinated and mounted on a Teflon membrane in Halocarbon oil 700 (Halocarbon Products Corp., Series 700, 9002-83-9). Images were acquired every 30 or 60 sec as indicated, single z = 0.5μm 18-20mm total on an upright Leica SP5 laser scanning confocal microscope equipped with a 63X 1.4 NA HCX PL Apochromat oil objective and LAS AF 2.2 software. Maximum intensity projections of confocal Z-stacks were rendered using Volocity visualization software (Improvision).

Embryos were collected, fixed in 4% PFA, and stained according to standard lab protocols [19 (link)]. Antibodies were used at the following concentrations: α-Dia (1:500) (S. Wasserman), α-Duf (1:200) (K.-F. Fischbach), α-Sns (1:250) (S. Abmayr), α-WASp (1:500) (E. Schejter), α-SCAR(1:100) (J Zallen) [85 (link)], α-Blow (1:100) (E. Chen), α-GFP (1:500)(Invitrogen A11120), α-dsRed (1: 500) (Clontech 32392), Alexa Fluor 647-phalloidin (1:100) (Invitrogen A22287). For secondary antibodies, Alexa Fluor 488-, Alexa Fluor 555-, and Alexa Fluor 647-conjugated fluorescent secondary antibodies at 1:200 dilution (Invitrogen) were used. Fluorescent images were acquired on a Leica SP5 laser scanning confocal microscope equipped with a 63X 1.4 NA HCX PL Apochromat oil objective and LAS AF 2.2 software. Maximum intensity projections of confocal Z-stacks were rendered using Volocity visualization software (Improvision). Cell outlines of somatic myoblasts and myotubes were determined in saturated images, and cells were false colored using Adobe Photoshop CS4 (S1 Fig).