TY - JOUR
T1 - A quantitative method to analyse F-actin distribution in cells
AU - Zonderland, Jip
AU - Wieringa, Paul
AU - Moroni, Lorenzo
N1 - Funding Information:
We are grateful to the European Research Council starting grant “Cell Hybridge” for financial support under the Horizon2020 framework program (Grant # 637308 ). Appendix A
Publisher Copyright:
© 2019 The Authors
PY - 2019
Y1 - 2019
N2 - 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 sizeAll steps in the analysis are shown, and can therefore easily be verified for each imageAll software required for the analysis is freely available (C) 2019 The Authors. Published by Elsevier B.V.
AB - 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 sizeAll steps in the analysis are shown, and can therefore easily be verified for each imageAll software required for the analysis is freely available (C) 2019 The Authors. Published by Elsevier B.V.
KW - Fiji macro
KW - Cytoskeleton distribution
KW - Automated image quantification
KW - Staining distribution
KW - Mesenchymal stromal cells
KW - Cytochalasin D
KW - OSTEOBLASTS
U2 - 10.1016/j.mex.2019.10.018
DO - 10.1016/j.mex.2019.10.018
M3 - Article
C2 - 31763187
VL - 6
SP - 2562
EP - 2569
JO - MethodsX
JF - MethodsX
ER -