Bellymount-pulsed tracking: a novel approach for real-time in vivo imaging of Drosophila abdominal tissues.

Quantitative live imaging is a valuable tool that offers insights into cellular dynamics. However, many fundamental biological processes are incompatible with current live-imaging modalities. Drosophila oogenesis is a well-studied system that has provided molecular insights into a range of cellular and developmental processes. The length of the oogenesis, coupled with the requirement for inputs from multiple tissues, has made long-term culture challenging. Here, we have developed Bellymount-pulsed tracking (Bellymount-PT), which allows continuous, noninvasive live imaging of Drosophila oogenesis inside the female abdomen for up to 16 h. Bellymount-PT improves upon the existing Bellymount technique by adding pulsed anesthesia with periods of feeding that support the long-term survival of flies during imaging. Using Bellymount-PT, we measure key events of oogenesis, including egg chamber growth, yolk uptake, and transfer of specific proteins to the oocyte during nurse cell dumping with high spatiotemporal precision within the abdomen of a live female.