Ixon 897 emccd

The light microscopy data on living tissue samples were taken using an Andor spinning-disk confocal microscope system equipped with an inverted microscope (Olympus; IX73), an iXon 897 EMCCD, and a 100×UPlanSApo objective (NA 1.40; Olympus) at 25°C. The acquisition software was Andor IQ 3.0. Photobleaching was performed using the Andor Frappa Unit coupled to the spinning disk microscope. The fluorescence recovery (R_rec, in percentage) was calculated as $R_{rec}=\frac{F_{\mathrm{t}}-F_0}{F_{-1}-F_0}\times 100\%$ where F₋₁ is the fluorescence signal in the region of interest before bleaching, F₀ is the signal right after bleaching, and F_t is the recovered signal after a period of time (t).

To image the expression pattern of DCX-EMAP, the tissue samples were imaged using a laser-scanning confocal microscope (LSM 780; Zeiss) equipped with a 63× oil UV-VIS-IR Apochromat objective (NA 1.2) at 25°C. To define the localization of DCX-EMAP, the fly halteres were imaged using a superresolution microscope (LSM980; Zeiss, Airyscan 2) equipped with a 63× oil Plan-Apochromat objective (NA 1.4; optical resolution: 120 nm) at 25°C. All the other optical microscopy data were collected using an Andor spinning disk confocal microscope (Andor) equipped with an inverted microscope (IX73; Olympus), an iXon 897 EMCCD, and a 100× UPlanSApo objective (NA 1.40; Olympus) at 25°C. The image acquisition software was Andor IQ 3.0 (Andor). The photobleaching experiments were performed using the Andor Frappa Unit (Andor) coupled to the spinning disk microscope. The fluorescence recovery (R_rec, in percentage) was calculated as $R_{rec}=\frac{F_{\mathrm{t}}-F_0}{F_{-1}-F_0}\times 100\%\text{,}$ where F_-1 is the fluorescence signal of the target region before bleaching, F₀ is the signal right after bleaching, and F_t is the recovered signal after a period (t).