Lucplanfl n 20 0.45 objective

Microscopy was performed using an Olympus BX61 VS120-S5 Virtual Slide Scanning System with UPlanSApo 40×/0.95 objective, Olympus XM10 monochrome camera, and Olympus VS-ASW FL 2.7 imaging software. Additionally, images were acquired using an Olympus FV1000 Confocal Microscope with UPlanFLN 40×/1.30 oil objective and Olympus FV-ASW version 4.2 imaging software. Live cell culture imaging was performed using an Olympus IX81 with LUCPlan FLN 20×/0.45 objective, Olympus XM10 monochrome camera and Olympus CellSens 1.13 software. Analysis and quantification were performed using Olympus CellSens 1.13, MetaMorph Premier 7.7.0.0, Adobe Photoshop CS6, and ImageJ software.

Male M. robinsoni (Otto and Hill, 2012) and M. nigromaculatus (Keyserling, 1883) were locally captured in New South Wales (M. robinsoni) and Queensland (M. nigromaculatus), Australia. All specimens were preserved in 70% ethanol. Details of both species' distribution can be found in ref. 6, 8 and 9 . Survey images of the scale organization at the opisthosomal flaps were made with an Olympus SZX16 stereomicroscope (Olympus, Tokyo, Japan) or a Zeiss Universal Microscope (Zeiss, Oberkochen, Germany) equipped with an Olympus LUCPlanFL N 20×/0.45 objective. All measurements were performed on at least two different specimens.

Reflectance spectra of single scales and scale-less wing areas were measured with a microspectrophotometer (MSP), consisting of a Leitz Ortholux microscope with a LUCPlanFL N 20×/0.45 objective (Olympus) and an Avantes AvaSpec-2048-2 CCD detector array spectrometer (Avantes, Apeldoorn, The Netherlands), with a xenon lamp light source. The reference was a white diffuse standard (Avantes WS-2). Wing reflectance spectra were measured with an integrating sphere and a bifurcated probe connected to a halogen/deuterium light source and the Avantes spectrometer. Reflectance spectra of the intact wing were also measured as a function of angle of light incidence for both transverse electric (TE)- and transverse magnetic (TM)-polarized light (where light is polarized perpendicular and parallel to the plane of light incidence, respectively) in a goniometric setup with two rotatable optical fibers. One fiber delivered light from a xenon lamp to the object, and the other fiber collected the reflected light and guided it to the spectrometer. The angular resolution of the setup has a Gaussian shape with half-width ∼5 deg (Stavenga et al., 2011 (link)). Naturally, the measured spectra slightly varied in shape and magnitude. The data in Figs 3–5 are representative, single cases.