Detection, Purification, Characterization, and Manipulation of Migrasomes.

Migrasomes are newly discovered cellular organelles, first described in 2015, that are formed by migrating cells. During migration, cells leave behind long membrane tethers called retraction fibers. Numerous micrometer-scale vesicles grow from the tips or junctions of these fibers, which we have named migrasomes. Migrasomes play important roles in various physiological processes, including releasing signaling factors to ensure organ morphogenesis during zebrafish embryo development, transferring mRNAs among cells, disposing of damaged mitochondria from the cell to maintain cell homeostasis, and secreting pro-angiogenic molecules to promote angiogenesis during chicken embryo development. Migrasomes are beginning to attract the attention of researchers in multiple fields. Here, we summarize the most commonly used protocols for migrasome detection using fluorescence microscopy imaging, purification through density-gradient centrifugation, characterization using electron microscopy (EM) imaging and biochemical analysis, and manipulation of migrasomes by targeting integrins and tetraspanins. © 2023 Wiley Periodicals LLC. Basic Protocol 1: Detection and observation of migrasomes by fluorescence microscopy imaging Basic Protocol 2: Purification of migrasomes from cultured cell lines and embryos by density-gradient centrifugation Basic Protocol 3: Characterization of migrasomes by electron microscopy imaging and biochemical analysis.