Vs asw fl

A FT from the temporal region (Fig. 1C) of a male Calumma crypticum (ZSM 503/2014, stored in 70% ethanol) was dissected with a razorblade. The excised FT was washed in 0.1 M phosphate buffer, stained in buffered 1% Osmium tetroxide (OsO₄) for one hour on ice, dehydrated in a graded acetone series, embedded in epoxy resin^{44 (link)}, serial sectioned in 279 planes à 1.54 µm using a RMC MT-7000 ultramicrotome with a Diatome Histo Jumbo diamond knife, mounted on glass slides, and partly stained with a 1:1 mixture of methylene blue and Azure II for approx. 5 s at 80 °C^{45 (link)}. Unstained sections (Supplementary Fig. S5A,B) were sealed under coverslips with DPX mounting medium (Agar Scientifics). The glass slides with slice-ribbons were imaged in bright field illumination using an Olympus BX61VS light microscope with UPlanSApo 10 × NA 0.4 objective and CX10 digital camera, and the program VS-ASW FL (Olympus v 2.7) for virtual slide acquisition. Images of single slices were then extracted with OlyVIA software (Olympus v 2.9; 1267 × 2119 px, 24 bit RGB, 0.98 µm/px) for subsequent volume rendering. In addition selected slices were photographed using an Olympus CX 41 light microscope with a DP25 digital camera (objectives: Olympus Plan C 10 × NA 0.25, Olympus UPlanSApo 40 × NA 0.95, and Olympus UPlanSApo 60 × NA 1.2 W); for details see Supplementary Table S8.