A quantitative method to analyse F-actin distribution in cells

Jip Zonderland*, Paul Wieringa, Lorenzo Moroni

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

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Abstract

Changes in actin structure and distribution are involved in many cellular processes, such as differentiation, proliferation and migration. Differences in cell shape and size make the analysis of actin distribution difficult. Here, we have developed a Fiji macro that analyzes the distribution of actin within the cell, regardless of cell size or shape. The staining intensity is measured along an automatically drawn line over the cell. The intensity data is divided in equal bins, making the analysis insensitive to changes in cell size or shape. We have also created an R script that further processes the acquired data. Together, final data can be acquired within minutes from a set of images, with freely available software. We demonstrate our method with F-actin staining of cytochalasin D treated cells. The advantages of our methods are:

The analysis is not influenced by cell shape or size

All steps in the analysis are shown, and can therefore easily be verified for each image

All software required for the analysis is freely available (C) 2019 The Authors. Published by Elsevier B.V.

Original languageEnglish
Pages (from-to)2562-2569
Number of pages8
JournalMethodsX
Volume6
DOIs
Publication statusPublished - 2019

Keywords

  • Fiji macro
  • Cytoskeleton distribution
  • Automated image quantification
  • Staining distribution
  • Mesenchymal stromal cells
  • Cytochalasin D
  • OSTEOBLASTS

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