RT Journal Article
SR Electronic
T1 A novel Fiji/ImageJ plugin for the rapid analysis of blebbing cells
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2021.06.04.447112
DO 10.1101/2021.06.04.447112
A1 Karl W. Vosatka
A1 Sandrine B. Lavenus
A1 Jeremy S. Logue
YR 2022
UL http://biorxiv.org/content/early/2022/02/25/2021.06.04.447112.abstract
AB When confined, cells have recently been shown to undergo a phenotypic switch to what has been termed, fast amoeboid (leader bleb-based) migration. However, as this is a nascent area of research, few tools are available for the rapid analysis of cell behavior. Here, we demonstrate that a novel Fiji/ImageJ-based plugin, Analyze_Blebs, can be used to quickly obtain cell migration parameters and morphometrics from time lapse images. As validation, we show that Analyze_Blebs can detect significant differences in cell migration and morphometrics, such as the largest bleb size, upon introducing different live markers of F-actin, including F-tractin and LifeAct tagged with green and red fluorescent proteins. We also demonstrate, using flow cytometry, that live markers increase total levels of F-actin. Furthermore, that F-tractin increases cell stiffness, which was found to correlate with a decrease in migration, thus reaffirming the importance of cell mechanics as a determinant of Leader Bleb-Based Migration (LBBM).Highlight summary A new plugin, Analyze_Blebs, enables the rapid analysis of cell migration and morphometrics of fast amoeboid cells. Morphometrics combined with cell stiffness are found to be predictive of confined, fast amoeboid (leader bleb-based), migration.Competing Interest StatementThe authors have declared no competing interest.