Protocol for acute oxidative stress induction in third-instar Drosophila melanogaster larvae

Here, we present a protocol for inducing oxidative stress in the Drosophila melanogaster larval brain to facilitate the screening of regulators involved in redox homeostasis. We describe steps for the collection of synchronized larvae and their treatment with oxidative stressors. We then detail procedures for larval brain dissection, lipid droplet staining, image acquisition, and the assessment of glutathione redox status. For complete details on the use and execution of this protocol, please refer to Li et al.¹.