Teneo lv sem

Individual scales from wild-type and mutant regions of interest were collected by brushing the surface of the wing with an eyelash tool, then dusted onto an SEM stub with double-sided carbon tape. Stubs were then colour imaged under the Keyence VHX-5000 microscope for registration of scale type. Samples were sputter-coated with one 12.5 nm layer of gold for improving sample conductivity. SEM images were acquired on a FEI Teneo LV SEM, using secondary electrons and an Everhart-Thornley detector using a beam energy of 2.00 kV, beam current of 25 pA, and a 10 μs dwell time. Individual images were stitched using the Maps 3.10 software (ThermoFisher Scientific).

Microstructural data were collected using a combination of advanced electron microscopy techniques. Secondary electron (SE) SEM images were acquired from MAC-etched shards of metallurgical and single-crystal electronics grade Si using an FEI Verios 460L SEM operating at an accelerating voltage of 2 kV. MAC-etched Si wafers were examined in an FEI Teneo LVSEM using an accelerating voltage of 5 kV. SE SEM images were acquired from cleaved samples to reveal the macropore geometry in cross-section. Cross-sectional TEM samples were produced from the MAC-etched silicon wafers using focused ion beam (FIB) techniques in an FEI Helios NanoLab 460F1 dual-beam FIB-SEM. TEM lamellae were prepared from the macropore by depositing a Pt layer in-situ to protect the near-surface region during Ga⁺ ion milling. Parallel trenches were then milled on either side of the Pt strap to define a pre-thinned lamella. The lamella was then transferred to a Cu Omni grid using a micro-manipulator needle; final thinning was performed at 30 kV. The FIB lift-outs were then analyzed in an FEI Talos F200X scanning transmission electron microscopy (STEM) operating at an accelerating voltage of 200 kV.