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METHODS IN CELL PHYSIOLOGY

Mechanical anisotropy of adherent cells probed by a three-dimensional magnetic twisting device

¹Physiology Program, Harvard School of Public Health, Boston 02115; ²Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts 02138; and ³EOL Eberhard, CH-4104 Oberwil, Switzerland

Submitted 6 May 2004 ; accepted in final form 16 June 2004

We describe a three-dimensional magnetic twisting device that is useful in characterizing the mechanical properties of cells. With the use of three pairs of orthogonally aligned coils, oscillatory mechanical torque was applied to magnetic beads about any chosen axis. Frequencies up to 1 kHz could be attained. Cell deformation was measured in response to torque applied via an RGD-coated, surface-bound magnetic bead. In both unpatterned and micropatterned elongated cells on extracellular matrix, the mechanical stiffness transverse to the long axis of the cell was less than half that parallel to the long axis. Elongated cells on poly-L-lysine lost stress fibers and exhibited little mechanical anisotropy; disrupting the actin cytoskeleton or decreasing cytoskeletal tension substantially decreased the anisotropy. These results suggest that mechanical anisotropy originates from intrinsic cytoskeletal tension within the stress fibers. Deformation patterns of the cytoskeleton and the nucleolus were sensitive to loading direction, suggesting anisotropic mechanical signaling. This technology may be useful for elucidating the structural basis of mechanotransduction.

cytoskeleton; prestress; stress fibers; mechanotransduction; mechanical deformation

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