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.
Highlights
In the last decades studies have shown the essential role of cell adhesion in processes like cell migration [1], survival, proliferation, and differentiation [2], as well as tissue morphogenesis [3]
focal adhesions (FAs) consist of a large number of proteins, such as vinculin, paxillin, focal adhesion kinase (FAK) and talin, and can range from 0.2 μm to 30 μm in size depending on the maturation stage of the FA as well as the cell type [5]
The robustness and applicability of the algorithm was tested by comparing three well-known factors affecting cellular, nuclear, and focal adhesion morphology to see whether the algorithm was able to detect changes in morphological features
Summary
In the last decades studies have shown the essential role of cell adhesion in processes like cell migration [1], survival, proliferation, and differentiation [2], as well as tissue morphogenesis [3]. These types of cell behavior are affected by the physical properties from the cell microenvironment as adherent cells have the ability to sense and respond to these properties by adapting their shape and orientation. Signals from the micro-environment are transmitted to the interior of the cell through a structural pathway, i.e. focal adhesions (FAs) physically linking the environment via the actin cytoskeleton to the nucleus. Diverse types of adhesion structures can be present, including small, round nascent focal adhesion structures
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