Ovaries were dissected in S2 media warmed to room temperature and fixed in PBS +0.1% Triton + 4% EM-grade formaldehyde (Polysciences). For actin staining only, ovaries were washed 3x in PBS + 0.1% Triton (PBT) then egg chambers were disrupted from the muscle sheath by gentle pipetting. Staining was performed with rhodamine phalloidin (1:200, Sigma), AlexaFluor-488 Phalloidin (1:200, Invitrogen) or AlexaFluor-647 Phalloidin (1:75, Invitrogen) for 15 minutes in PBT at room temperature while rocking. Egg chambers were then washed 3x in PBT and mounted in SlowFade Antifade (Invitrogen). Egg chambers stained with primary antibodies were incubated overnight at 4°C with rocking in PBT + 0.1% BSA + Phalloidin. Egg chambers were incubated in AlexaFluor-488 or 555 conjugated secondary antibodies (1:200, Invitrogen) in PBT + 0.1% BSA + Phalloidin for 3 hours at room temperature with rocking. The following primary antibodies were used: anti-Ena (1:200 concentrate, DHSB), anti-SCAR (1:300)43 (link), and anti-SCAR (1:200 concentrate, DSHB)44 (link). For SCAR staining, primary and secondary incubations along with all washes following the primary incubation, PBT3 (PBS + 0.3% Triton) was used instead of PBT. Fixed egg chambers were imaged with a laser-scanning confocal or a spinning disk confocal (described above). ImageJ and Adobe Photoshop were used for image processing.