Stiffness tomography of eukaryotic intracellular compartments by atomic force microscopy

Abstract

After identification by fluorescence microscopy, intracellular compartments are analyzed by stiffness tomography using atomic force microscopy, before further processing for ultrastructural characterization by electron microscopy.

Subsurface Imaging of Cell Organelles by Force Microscopy

“… 42 , 43 This method has been applied by Lafont and co-workers to image a cell infected by a bacteria. 44 …”

The use of nanovibration to discover specific and potent bioactive metabolites that stimulate osteogenic differentiation in mesenchymal stem cells

“…Keeping the maximum indentation used in the calculation under ~10-15% of the thickness, and 20-25% of the radius of the indenter, is a rule of thumb typically used in literature (in our case, 600nm-700nm would match these requirements). Nevertheless, for indentations lower than this threshold, a trend in the measured Young's modulus as a function of the indentation depth appears, which is associated with the inhomogeneity of the cell 62 . Here,we exploited this approach, trying to isolate the elasticity of the cortical region from the bulk of the cell.…”

Cells nanomechanics by atomic force microscopy: focus on interactions at nanoscale

“…Finally, relative to the vertical heterogeneity of living cells we point out the so-called stiffness tomography obtained by sectioning the indentation curve and fitting the modulus with standard contact models, evidencing the presence of stiffer cytoskeletal bundles beneath cell surface [50][51][52]. Contrast by stiffness tomography was used to visualize and obtain mechanical feedback about mitochondria and Golgi apparatus without specific staining [53]. Rheological model was applied on stiffness tomography for fibroblast cells localizing actin bundle fibers under cell surface or nucleoli within nucleus structure from the contrast of different hydrodynamic drag [54].…”