A Protocol for Measuring Plasma Membrane Tension in the Drosophila Ovary Using Fluorescence Lifetime Imaging Microscopy

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