Fig. 10
Detecting BTV infection in single planes of salivary apparatus of female Culicoides sonorensis. Example of single planes (z1 to z3) for two of the insects shown in Fig. 5. Viral presence and replication in the main lobe can be missed when analyzing single planes/sections due to the high ratio of lumen:cellular structures. Equally, accessory glands are hard to identify. In midge 1, cellular tubulin is shown in blue (labelling with mouse anti-tubulin, and anti-mouse IgG AlexaFluor™ 405), BTV viral structural proteins (VSPs) in green (labelling with in-house guinea pig antibody Orab279 and anti-guinea pig IgG AlexaFluor™ 488), and BTV non-structural protein 2 (NS2) in red (labelling with in-house rabbit antibody Orab1 and anti-rabbit IgG AlexaFluor™ 568). In midge 2, cell nuclei are shown in blue (DAPI staining), cellular tubulin is visualised in red (labelling with mouse anti-tubulin, and anti-mouse IgG AlexaFluor™ 568) and BTV VSPs are visualized in green (labelling with in-house antibody Orab279 and anti-guinea pig IgG AlexaFluor™ 488). In all panels, microscope magnification is shown on the top right corner and the scale bar represents 20 µm