Fig. 8
Absence of BTV infection in salivary apparatus of Culicoides nubeculosus fed with an infectious bloodmeal. Three representative images of salivary apparatus from three different females of C. nubeculosus at 8 days post infection. Images on the left show one plane taken with differential interference contrast (DIC) microscopy. Images on the right are maximum projection of at least 50 stack images. In midge 1 and 3, 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). BTV proteins were not observed in any of the midges. In midge 1, four accessory glands around one main salivary gland are numbered and the ampulla (oval) and salivary duct (arrow) can be observed. In all panels, microscope magnification is shown on the top right corner and the scale bar represents 20 µm