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A novel Fiji/ImageJ plugin for the rapid analysis of blebbing cells

Karl W. Vosatka, Sandrine B. Lavenus, Jeremy S. Logue
doi: https://doi.org/10.1101/2021.06.04.447112
Karl W. Vosatka
Department of Regenerative and Cancer Cell Biology, Albany Medical College, 47 New Scotland Ave, Albany, NY 12208
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Sandrine B. Lavenus
Department of Regenerative and Cancer Cell Biology, Albany Medical College, 47 New Scotland Ave, Albany, NY 12208
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Jeremy S. Logue
Department of Regenerative and Cancer Cell Biology, Albany Medical College, 47 New Scotland Ave, Albany, NY 12208
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  • For correspondence: loguej@amc.edu
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Abstract

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 Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted February 25, 2022.
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A novel Fiji/ImageJ plugin for the rapid analysis of blebbing cells
Karl W. Vosatka, Sandrine B. Lavenus, Jeremy S. Logue
bioRxiv 2021.06.04.447112; doi: https://doi.org/10.1101/2021.06.04.447112
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A novel Fiji/ImageJ plugin for the rapid analysis of blebbing cells
Karl W. Vosatka, Sandrine B. Lavenus, Jeremy S. Logue
bioRxiv 2021.06.04.447112; doi: https://doi.org/10.1101/2021.06.04.447112

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