Atomic force microscopy (AFM) has emerged as a promising tool to characterize the mechanical properties of biological materials and cells. In our studies, undifferentiated and early differentiating mouse embryonic stem cells (mESCs) were assessed individually using an AFM system to determine if we could detect changes in their mechanical properties by surface probing. Probes with pyramidal and spherical tips were assessed, as were different analytical models for evaluating the data. The combination of AFM probing with a spherical tip and analysis using the Hertz model provided the best fit to the experimental data obtained and thus provided the best approximation of the elastic modulus. Our results showed that after only 6 days of differentiation, individual cell stiffness increased significantly with early differentiating mESCs having an elastic modulus two-to threefold higher than undifferentiated mESCs, regardless of cell line (R1 or D3 mESCs) or treatment. Singletouch (indentation) probing of individual cells is minimally invasive compared to other techniques. Therefore, this method of mechanical phenotyping should prove to be a valuable tool in the development of improved methods of identification and targeted cellular differentiation of embryonic, adult, and induced-pluripotent stem cells for therapeutic and diagnostic purposes.
International audienceIn order to understand and characterize the mechanical property and response of the mouse embryonic stem cells (mESC), we used an atomic force microscope (AFM) combined with a PHANToM haptic feedback device. Atomic force microscopy has rapidly become a valuable tool for quantifying the biophysical properties of single cells or a collection of cells through force measurements. We report herein the mechanical characterization of single mESC using indentation-relaxation measurements with micro-sphere AFM probes for fixed and live undifferentiated mESC. During cell indentation for both live and fixed undifferentiated cells, we provided force feedback to the user in real-time through the PHANToM haptic feedback device as the AFM tip was deforming the cell. The force was amplified for the human operator to perceive the change in force during cell indentation by the AFM cantilever. This information can be used as a mechanical marker to characterize state of the cell (live and fixed). As the interpretation of atomic force microscopy-based indentation tests is highly dependent on the use of an appropriate theoretical model of the testing configuration, various contact models are presented to predict the mechanical behavior of an individual mouse embryonic stem cells (mESC) in different states. A comparison study with finite element simulations (FEM) of spherical tip indentation demonstrates the effectiveness of our computational model to predict the mESC deformation during indentation and relaxation nanomanipulation tasks
International audienceTraining simulators that provide realistic visual and haptic feedback during cell indentation tasks are currently inves tigated. Complex cell geometry inherent to biological cells and intricate mechanical properties drive the need for precise mechan ical and numerical modeling to assure accurate cell deformation and force calculations. Advances in alternative finite-element for mulation, such as the mass-tensor approach, have reached a state, where they are applicable to model soft-cell deformation in real time. The geometrical characteristics and the mechanical proper ties of different cells are determined with atomic force microscopy (AFM) indentation. A real-time, haptics-enabled simulator for cell centered indentation has been developed, which utilizes the AFM data (mechanical and geometrical properties of embryonic stem cells) to accurately replicate the indentation task and predict the cell deformation during indentation in real time. This tool can be used as a mechanical marker to characterize the biological state of the cell. The operator is able to feel the change in the stiff ness during cell deformation between fixed and live cells in real time. A comparative study with finite-element simulations using a commercial software and the experimental data demonstrate the effectiveness of the proposed physically based model
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