Protocol to chemically deplete phagocytic hemocytes in Anopheles gambiae using clodronate liposomes

Understanding the roles of phagocytic hemocytes in mosquito innate immunity has been significantly limited due to the lack of genetic tools. Here, we present a protocol for depleting phagocytic hemocytes in Anopheles gambiae mosquitoes using clodronate liposomes. We describe steps for mosquito injection, as well as validation by microscopy, quantitative real-time PCR (real-time qPCR), and flow cytometry analysis. This protocol allows for the delineation of phagocytic hemocyte function in mosquito immunity, which can be more broadly applied to other arthropod systems. For complete details on the use and execution of this protocol, please refer to Kwon et al.¹.