Methods in Anopheles Research - MR4

Chapter 6 : Dissection Techniques6.5 Dissecting Plasmodium-Infected Mosquitoes6.5.2 Salivary GlandsPage 2 of 2StainingLet glands fully dry on the slide. Once dried, fix with methanol for 1 minute, rinse with water, then stainwith Giemsa stain for 40 minutes. Rinse again and air dry before inspecting under a microscope.What you will seeIn uninfected salivary glands, you will see a tissue layer with no visible threadlike sporozoites (Figure6.5.2.2). In infected mosquitoes, you will see the threadlike parasites both in the tissues or, if the gland isruptured, in the field surrounding the tissues (Figure 6.5.2.3). Giemsa staining helps estimate lowparasite loads by increasing resolution of the sporozoites within the glands.Figure 6.5.2.1. Dissection of salivary glands.Figure 6.5.2.2. Stained, uninfected salivaryglands.Figure 6.5.2.3. Stained, infected salivaryglands. Note the thin, threadlike sporozoites(circled) of P. vivax spilling from the salivarygland.ReferencesLooker M, Taylor-Robinson AW (2004) Methods in Malaria Research, 4th edn. MR4/ATCC, Manassas,VAWHO (1975) Manual on practical entomology in malaria. Part II. In. World Health Organization, Geneva, p191pp.