Measuring Honeybee and Wasp Head Morphology

Heads were dissected from honeybee pupae and photographed using an S8 APO binocular microscope (Leica, Wetzlar, Germany) with a UI-1240LE-C-HQ camera (IDS, Obersulm, Germany) and uEye Cockpit (part of IDS suite v4.92, IDS, Obersulm, Germany) software. Images for dorsal lens facet diameter measurements were taken with a Canon Eos 6d Mark II camera and Canon MP-E65 mm 1:2,8 1–5× Marco objective (Tokyo, Japan). Images of N. vitripennis adult heads were obtained using a Dino-Lite Edge 5MP Digital Microscope and DinoCapture 2.0 software (Dino-Lite, Almere, The Netherlands). Head parameters (head width, head length, eye width, eye length and interocular distance) were measured as schematically presented (Supplementary Fig. 11). In glu^{ex7-8 F} males and controls, lens facet diameter was measured in the frontal-dorsal region of the eye, where facets in males should be largest^{24 (link)}. The mean of 15 randomly measured lens facet diameters per individual was used for analysis. Length measures were performed using ImageJ (National Institute of Mental Health, USA).