The mechanical properties of cells influence their cellular and subcellular functions, including cell adhesion, migration, polarization, and differentiation, as well as organelle organization and trafficking inside the cytoplasm. Yet reported values of cell stiffness and viscosity vary substantially, which suggests differences in how the results of different methods are obtained or analyzed by different groups. To address this issue and illustrate the complementarity of certain approaches, here we present, analyze, and critically compare measurements obtained by means of some of the most widely used methods for cell mechanics: atomic force microscopy, magnetic twisting cytometry, particle-tracking microrheology, parallel-plate rheometry, cell monolayer rheology, and optical stretching. These measurements highlight how elastic and viscous moduli of MCF-7 breast cancer cells can vary 1,000-fold and 100-fold, respectively. We discuss the sources of these variations, including the level of applied mechanical stress, the rate of deformation, the geometry of the probe, the location probed in the cell, and the extracellular microenvironment.

The atomic force microscope is broadly used to study the morphology of cells1–5 but it can also probe the mechanics of cells. It is now known that cancerous cells may have different mechanical properties than normal cells6–8 but the reasons for these differences are poorly understood9. Here we report quantitatively the differences between normal and cancerous human cervical epithelial cells by considering the brush layer on the cell surface. These brush layers, which consist mostly of microvilli, microridges, and cilia are important for interacting with the environment. Deformation force curves obtained from cells in vitro are processed according to the 'brush on soft cell model 10. We found that normal cells have brushes with one length while cancerous cells displayed long and short brushes with significantly different densities. The observed differences suggest that brush layers should be taken into account when characterizing the cell surface by mechanical means.

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Here we investigated the question whether cells, being highly heterogeneous objects, could be described with the elastic modulus (effective Young's modulus) in a self-consistent way. We performed a comparative analysis of the elastic modulus derived from the indentation data obtained with atomic force microscopy (AFM) on human cervical epithelial cells (both normal and cancerous). Both sharp (cone) and dull (2500-nm radius sphere) AFM probes were used. The indentation data were processed through different elastic models. The cell was approximated as a homogeneous elastic medium that had either 1), smooth hemispherical boundary (Hertz/Sneddon models) or 2), the boundary covered with a layer of glycocalyx and membrane protrusions ("brush" models). Consistency of these approximations was investigated. Specifically, we tested the independence of the elastic modulus of the indentation depth, which is assumed in these models. We demonstrated that only one model showed consistency in treating cells as a homogeneous elastic medium, namely, the brush model, when processing the indentation data collected with the dull AFM probe. The elastic modulus demonstrated strong depth dependence in all models: Hertz/Sneddon models (no brush taken into account), and when the brush model was applied to the data collected with sharp conical probes. We conclude that it is possible to describe the elastic properties of the cell body by means of an effective elastic modulus, used in a self-consistent way, when using the brush model to analyze data collected with a dull AFM probe. The nature of these results is discussed.

The modulus of elasticity of soft materials on the nanoscale is of interest when studying thin films, nanocomposites, and biomaterials. Two novel modes of atomic force microscopy (AFM) have been introduced recently: HarmoniX and PeakForce QNM. Both modes produce distribution maps of the elastic modulus over the sample surface. Here we investigate the question of how quantitative these maps are when studying soft materials. Three different polymers with a macroscopic Young's modulus of 0.6-0.7 GPa (polyurethanes) and 2.7 GPa (polystyrene) are analyzed using these new modes. The moduli obtained are compared to the data measured with the other commonly used techniques, dynamic mechanical analyzer (DMA), regular AFM, and nanoindenter. We show that the elastic modulus is overestimated in both the HarmoniX and PeakForce QNM modes when using regular sharp probes because of excessively overstressed material in the samples. We further demonstrate that both AFM modes can work in the linear stress-strain regime when using a relatively dull indentation probe (starting from ~210 nm). The analysis of the elasticity models to be used shows that the JKR model should be used for the samples considered here instead of the DMT model, which is currently implemented in HarmoniX and PeakForce QNM modes. Using the JKR model and ~240 nm AFM probe in the PeakForce QNM mode, we demonstrate that a quantitative mapping of the elastic modulus of polymeric materials is possible. A spatial resolution of ~50 nm and a minimum 2 to 3 nm indentation depth are achieved.