An automated quantitative analysis of cell, nucleus and focal adhesion morphology

PLoS One. 2018 Mar 30;13(3):e0195201. doi: 10.1371/journal.pone.0195201. eCollection 2018.

Abstract

Adherent cells sense the physical properties of their environment via focal adhesions. Improved understanding of how cells sense and response to their physical surroundings is aided by quantitative evaluation of focal adhesion size, number, orientation, and distribution in conjunction with the morphology of single cells and the corresponding nuclei. We developed a fast, user-friendly and automated image analysis algorithm capable of capturing and characterizing these individual components with a high level of accuracy. We demonstrate the robustness and applicability of the algorithm by quantifying morphological changes in response to a variety of environmental changes as well as manipulations of cellular components of mechanotransductions. Finally, as a proof-of-concept we use our algorithm to quantify the effect of Rho-associated kinase inhibitor Y-27632 on focal adhesion maturation. We show that a decrease in cell contractility leads to a decrease in focal adhesion size and aspect ratio.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Algorithms
  • Amides / pharmacology
  • Automation
  • Cell Nucleus / drug effects
  • Cell Nucleus / metabolism*
  • Dose-Response Relationship, Drug
  • Focal Adhesions* / drug effects
  • Humans
  • Image Processing, Computer-Assisted
  • Molecular Imaging
  • Pyridines / pharmacology
  • rho-Associated Kinases / antagonists & inhibitors

Substances

  • Amides
  • Pyridines
  • Y 27632
  • rho-Associated Kinases

Grants and funding

This work was supported by CVON 2012-01 Dutch Heart Foundation Antonetta B. C. Buskermolen and TU/e Impuls Antonetta B. C. Buskermolen. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.