STRESS, an automated geometrical characterization of deformable particles for in vivo measurements of cell and tissue mechanical stresses

Abstract

From cellular mechanotransduction to the formation of embryonic tissues and organs, mechanics has been shown to play an important role in the control of cell behavior and embryonic development. Most of our existing knowledge of how mechanics affects cell behavior comes from in vitro studies, mainly because measuring cell and tissue mechanics in 3D multicellular systems, and especially in vivo, remains challenging. Oil microdroplet sensors, and more recently gel microbeads, use surface deformations to directly quantify mechanical stresses within developing tissues, in vivo and in situ, as well as in 3D in vitro systems like organoids or multicellular spheroids. However, an automated analysis software able to quantify the spatiotemporal evolution of stresses and their characteristics from particle deformations is lacking. Here we develop STRESS (Surface Topography Reconstruction for Evaluation of Spatiotemporal Stresses), an analysis software to quantify the geometry of deformable particles of spherical topology, such as microdroplets or gel microbeads, that enables the automatic quantification of the temporal evolution of stresses in the system and the spatiotemporal features of stress inhomogeneities in the tissue. As a test case, we apply these new code to measure the temporal evolution of mechanical stresses using oil microdroplets in developing zebrafish tissues. Starting from a 3D timelapse of a droplet, the software automatically calculates the statistics of local anisotropic stresses, decouples the deformation modes associated with tissue- and cell-scale stresses, obtains their spatial features on the droplet surface and analyzes their spatiotemporal variations using spatial and temporal stress autocorrelations. The automated nature of the analysis will help users obtain quantitative information about mechanical stresses in a wide range of 3D multicellular systems, from developing embryos or tissue explants to organoids. Author summaryThe measurement of mechanical stresses in 3D multicellular systems, such as living tissues, has been very challenging because of a lack in technologies for this purpose. Novel microdroplet techniques enable direct, quantitative in situ measurements of mechanical stresses in these systems. However, computational tools to obtain mechanical stresses from 3D images of microdroplets in an automated and accurate manner are lacking. Here we develop STRESS, an automated analysis software to analyze the spatiotemporal characteristics of mechanical stresses from microdroplet deformations in a wide range of systems, from living embryonic tissues and tissue explants to organoids and multicellular spheroids.