Abstract
Mechanosensation of plasma membrane tension by various mechanoresponsive machineries is crucial for regulating stem cell fate, cell adhesion, and tissue morphogenesis. Here, we present a protocol for evaluating plasma membrane stretching during the differentiation of Drosophila ovarian cyst using a fluorescent lipid tension reporter (Flipper-TR). We describe the steps for microphone setup, ovary dissection, Flipper-TR staining, fluorescence lifetime imaging microscopy imaging, and image processing and analysis. This protocol demonstrates the utility of Flipper-TR for investigating the impact of mechanical forces in living tissue. For complete details on the use and execution of this protocol, please refer to Wang et al.1.