Practical aspects of the cellular force inference toolkit (CellFIT).

If we are to fully understand the reasons that cells and tissues move and acquire their distinctive geometries during processes such as embryogenesis and wound healing, we will need detailed maps of the forces involved. One of the best current prospects for obtaining this information is noninvasive force-from-images techniques such as CellFIT, the Cellular Force Inference Toolkit, whose various steps are discussed here. Like other current quasistatic approaches, this one assumes that cell shapes are produced by interactions between interfacial tensions and intracellular pressures. CellFIT, however, allows cells to have curvilinear boundaries, which can significantly improve inference accuracy and reduce noise sensitivity. The quality of a CellFIT analysis depends on how accurately the junction angles and edge curvatures are measured, and a software tool we describe facilitates determination and evaluation of this information. Special attention is required when edges are crenulated or significantly different in shape from a circular arc. Because the tension and pressure equations are overdetermined, a select number of edges can be removed from the analysis, and these might include edges that are poorly defined in the source image, too short to provide accurate angles or curvatures, or noncircular. The approach works well for aggregates with as many as 1000 cells, and introduced errors have significant effects on only a few adjacent cells. An understanding of these considerations will help CellFIT users to get the most out of this promising new technique.