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AFM for Cell Biology and Tissue Engineering

Atomic force microscopy image of a cell overlaid on a fluorescence microscopy image of the same cell

Atomic force microscopy is an essential tool for cell biology research. It can provide 3D topographical data of living, unfixed cells. However, AFM’s greatest strength in cell biology is its ability to provide accurate and quantitative mechanical measurements in near-physiological conditions (i.e. in culture medium and at 37°C). The elastic and viscoelastic response of a cell or substrate can be routinely measured using force maps and AFM-based microrheology techniques, respectively. The measured cell moduli can be that of unaltered cells, cells in different states of development, differentiation, or disease, or cells responding to a stimulus such as a drug or mechanical stress. Measuring the moduli of substrates and the cell microenvironment is also important due to the role the extracellular matrix (ECM) plays in such processes as cell differentiation, fate, signalling, gene transcription, cancer, cardiovascular disease and apoptosis.
 
When integrated with an inverted optical microscope (i.e. fluorescence, confocal, TIRF, etc.), data from both imaging modalities can be combined to correlate fluorescently labeled structures with AFM topography. The optics can be used to direct the AFM tip to probe a particular region of the cell, which can be crucial for hard-to-image cell types. Finally, AFM can also be used to provide a mechanical stimulus to cells and the associated response (e.g. ion handling, membrane potential changes, etc.) can be recorded optically in order to understand mechanotransduction in living cells and tissues.

Ask an AFM expert for more information
  • Image live cells in culture
  • Measure the elastic or viscoelastic response of cells and substrates
  • Integrate AFM with inverted optical microscopes and fluorescent techniques
  • Use optical images to select a region of interest (ROI) for AFM images and/or force measurements
  • Overlay AFM topography or modulus maps onto optical images and 3D AFM images
  • Dynamic imaging of living cells after treatment
  • Stiffness and viscoelastic changes in cancer cells
  • Influence of cell substrates on cell differentiation
  • Response of cells to mechanical stimulation

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M. F. Murphy, M. J. Lalor, F. C. Manning, F. Lilley, S. R. Crosby, C. Randall, and D. R. Burton, "Comparative study of the conditions required to image live human epithelial and fibroblast cells using atomic force microscopy," Microsc. Res. Tech. 69, 757-765 (2006). doi:10.1002/jemt.20339

M. Prabhune, G. Belge, A. Dotzauer, J. Bullerdiek, and M. Radmacher, "Comparison of mechanical properties of normal and malignant thyroid cells," Micron 43, 1267-1272 (2012). doi:10.1016/j.micron.2012.03.023

M. Prass, "Direct measurement of the lamellipodial protrusive force in a migrating cell," J. Cell Biol. 174, 767-772 (2006). doi:10.1083/jcb.200601159

A. Raman, S. Trigueros, A. Cartagena, A. P. Z. Stevenson, M. Susilo, E. Nauman, and S. A. Contera, "Mapping nanomechanical properties of live cells using multi-harmonic atomic force microscopy," Nat. Nanotechnol. 6, 809-814 (2011). doi:10.1038/nnano.2011.186

F. Rehfeldt, A. E. X. Brown, M. Raab, S. Cai, A. L. Zajac, A. Zemel, and D. E. Discher, "Hyaluronic acid matrices show matrix stiffness in 2D and 3D dictates cytoskeletal order and myosin-II phosphorylation within stem cells," Integr. Biol. 4, 422 (2012). doi:10.1039/c2ib00150k

J. Rother, H. Noding, I. Mey, and A. Janshoff, "Atomic force microscopy-based microrheology reveals significant differences in the viscoelastic response between malign and benign cell lines," Open Biology 4, 140046-140046 (2014). doi:10.1098/rsob.140046

S. Sen, S. Subramanian, and D. E. Discher, "Indentation and Adhesive Probing of a Cell Membrane with AFM: Theoretical Model and Experiments," Biophys. J. 89, 3203-3213 (2005). doi:10.1529/biophysj.105.063826

E. Spedden, and C. Staii, "Neuron Biomechanics Probed by Atomic Force Microscopy," Int. J. Mol. Sci. 14, 16124-16140 (2013). doi:10.3390/ijms140816124

E. Spedden, J. D. White, E. N. Naumova, D. L. Kaplan, and C. Staii, "Elasticity Maps of Living Neurons Measured by Combined Fluorescence and Atomic Force Microscopy," Biophys. J. 103, 868-877 (2012). doi:10.1016/j.bpj.2012.08.005

J. R. Tse, and A. J. Engler, "Stiffness Gradients Mimicking In Vivo Tissue Variation Regulate Mesenchymal Stem Cell Fate," PLoS ONE 6, e15978 (2011). doi:10.1371/journal.pone.0015978

K. R. Wilhelm, E. Roan, M. C. Ghosh, K. Parthasarathi, and C. M. Waters, "Hyperoxia increases the elastic modulus of alveolar epithelial cells through Rho kinase," FEBS Journal 281, 957-969 (2013). doi:10.1111/febs.12661

Y. Xiong, A. C. Lee, D. M. Suter, and G. U. Lee, "Topography and Nanomechanics of Live Neuronal Growth Cones Analyzed by Atomic Force Microscopy," Biophys. J. 96, 5060-5072 (2009). doi:10.1016/j.bpj.2009.03.032

W. Xu, R. Mezencev, B. Kim, L. Wang, J. McDonald, and T. Sulchek, "Cell Stiffness Is a Biomarker of the Metastatic Potential of Ovarian Cancer Cells," PLoS ONE 7, e46609 (2012). doi:10.1371/journal.pone.0046609

E. K. Yim, E. M. Darling, K. Kulangara, F. Guilak, and K. W. Leong, "Nanotopography-induced changes in focal adhesions, cytoskeletal organization, and mechanical properties of human mesenchymal stem cells," Biomaterials 31, 1299-1306 (2010). doi:10.1016/j.biomaterials.2009.10.037