Sz stereomicroscope

SGs collected from male Myd88^+/+ and Myd88^-/- mice (10 weeks of age) were immersed in periodate-lysine-paraformaldehyde-fixative for 6 h at 4°C, embedded in paraffin and serially sectioned at 5 µm of thickness. Sections were stained with hematoxylin and eosin and histologically analyzed. For immunofluorescent staining, the tissue sections were deparaffinized and immersed in distilled water. The sections were treated with 0.1% proteinase K in phosphate-buffered saline (PBS) for 5 min at room temperature and washed three times with PBS. The sections were then blocked with 1% bovine serum albumin (BSA) in PBS followed by incubation for 1 h at room temperature with rabbit anti-SLPI (secretory leukocyte peptidase inhibitor) antibody (OAPB00538; Aviva System Biology, San Diego, CA, USA). After washing three times with PBS, the sections were incubated for 30 min at room temperature with Alexa Fluor 488-conjugated anti-rabbit IgG antibody (Life Technologies, Rockville, MD, USA) and propidium iodide. After washing with PBS, the sections were sealed in the presence of Prolong Gold anti-fade reagent (Life Technologies). Optical and fluorescent images were obtained using an SZ stereomicroscope with DP21 digital camera (Olympus, Tokyo, Japan), a BX41 microscope (Olympus) and, a Biozero fluorescence microscope (KEYENCE, Osaka, Japan) and processed using Adobe Photoshop (Adobe, San Jose, CA).

Mosquito species identification was undertaken morphologically under a dissecting microscope. Before commencing any efficacy test of the selected larvicides against An. stephensi, 30 adult female mosquitoes were randomly aspirated from cages. Then these mosquitoes were transferred into a glass tube and exposed to chloroform by a cotton ball wetted at the tip. Each of these mosquitoes was laid under a Olympus SZ stereomicroscope at 40X for morphological identification using the updated key to the females of Afro-tropical Anopheles mosquitoes, which includes An. stephensi [15 (link)]. All were confirmed to be An. stephensi. Fewer Culex and Aedes larvae were collected compared to An. stephensi from the same habitats. Though there were a few Culex and Aedes species larvae collected with An. stephensi, all emerged adults aspirated from the cage were An. stephensi. Typical features of the morphology of An. stephensi are (i) the appearance with 3 pale bands in the palpus and the two apical pale bands are very broad with speckling on palpus segment 3 and (ii) in the 2nd main dark area on the vein 1of its wing, there are 2 pale interruptions [15 (link)].