Unmated and mated females were fixed for 1 h in 4% FA after the removal of legs and wings, washed 3 × 5 min in PBS (0.85% NaCl, 1.4 mM KH2 PO4, 8 mM Na2 HPO4, pH 7.4) and incubated in 25% sucrose overnight at 4 °C. Flies were briefly washed in PBS before embedding in O.C.T (#4583, Tissue-Tek) and freezing on a LN2 cooled metal block. Using a cryostat (Leica CM1950), 20 µm sections were collected on super plus glass slides (Thermo Scientific) and coated with 1 nm AuPd (Quorum Q 150 T ES, UK).
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