Quantitative Image Analysis of Dynamic Cell Behaviors During Border Cell Migration

Drosophila border cells have emerged as a genetically tractable model to investigate dynamic collective cell migration within the context of a developing organ. Studies of live border cell cluster migration have revealed similarities with other migrating collectives, including formation and restriction of cellular protrusions to the front of the cluster, supracellular actomyosin contractility of the entire collective, and intra-collective cell motility. Here, we describe protocols to prepare ex vivo cultures of stage 9 egg chambers followed by live time-lapse imaging of fluorescently labeled border cells to image dynamic cell behaviors. We provide options to perform live imaging using either a widefield epifluorescent microscope or a confocal microscope. We further outline steps to quantify various cellular behaviors and protein dynamics of live migrating border cells using the Fiji image processing package of ImageJ. These methods can be adapted to other migrating cell collectives in cultured tissues and organs.