Biomechanical imaging of cell stiffness and prestress with subcellular resolution

Abstract

Knowledge of cell mechanical properties, such as elastic modulus, is essential to understanding the mechanisms by which cells carry out many integrated functions in health and disease. Cellular stiffness is regulated by the composition, structural organization, and indigenous mechanical stress (or prestress) borne by the cytoskeleton. Current methods for measuring stiffness and cytoskeletal prestress of living cells necessitate either limited spatial resolution but with high speed, or spatial maps of the entire cell at the expense of long imaging times. We have developed a novel technique, called biomechanical imaging, for generating maps of both cellular stiffness and prestress that requires less than 30 s of interrogation time, but which provides subcellular spatial resolution. The technique is based on the ability to measure tractions applied to the cell while simultaneously observing cell deformation, combined with capability to solve an elastic inverse problem to find cell stiffness and prestress distributions. We demonstrated the application of this technique by carrying out detailed mapping of the shear modulus and cytoskeletal prestress distributions of 3T3 fibroblasts, making no assumptions regarding those distributions or the correlation between them. We also showed that on the whole cell level, the average shear modulus is closely associated with the average prestress, which is consistent with the data from the literature. Data collection is a straightforward procedure that lends itself to other biochemical/biomechanical interventions. Biomechanical imaging thus offers a new tool that can be used in studies of cell biomechanics and mechanobiology where fast imaging of cell properties and prestress is desired at subcellular resolution.