Measuring biological materials mechanics with atomic force microscopy ‐ Determination of viscoelastic cell properties from stress relaxation experiments

Cells are complex, viscoelastic bodies. Their mechanical properties are defined by the arrangement of semiflexible cytoskeletal fibers, their crosslinking, and the active remodeling of the cytoskeletal network. Atomic force microscopy (AFM) is an often‐used technique for the study of cell mechanics, enabling time‐ and frequency‐dependent measurements with nanometer resolution. Cells exhibit time‐dependent deformation when stress is applied. In this work, we have investigated the stress relaxation of HeLa cells when subjected to a constant strain. We have varied the applied force (1, 2, 4, and 8 nN) and pause time (1, 10, and 60 s) to check for common assumptions for the use of models of linear viscoelasticity. Then, we have applied three models (standard linear solid, five element Maxwell, power law rheology) to study their suitability to fit the datasets. We show that the five element Maxwell model captures the stress relaxation response the best while still retaining a low number of free variables. This work serves as an introduction and guide when performing stress relaxation experiments on soft matter using AFM. RESEARCH HIGHLIGHTS: Cells exhibit linear viscoelastic properties when subjected to stress relaxation measurements at the studied different forces and times. The stress relaxation is best described by a five element Maxwell model. All three used models capture a softening and fluidization of cells when disrupting actin filaments.