A Novel Fiji/ImageJ Plugin for the Rapid Analysis of Blebbing Cells

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2022 Apr 29

PMID 35486637

Authors

Karl W Vosatka

Sandrine B Lavenus

Jeremy S Logue

Affiliations

Soon will be listed here.

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).

Citing Articles

Structure of the F-tractin-F-actin complex.

Shatskiy D, Sivan A, Wedlich-Soldner R, Belyy A J Cell Biol. 2025; 224(4).

PMID: 39928047 PMC: 11809415. DOI: 10.1083/jcb.202409192.

The activation of INF2 by Piezo1/Ca is required for mesenchymal to amoeboid transition in confined environments.

Kar N, Caruso A, Prokopiou N, Logue J bioRxiv. 2023; .

PMID: 37745412 PMC: 10515767. DOI: 10.1101/2023.06.23.546346.

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