A new method for application of force to cells via ferric oxide beads.
We describe a new method that uses straightforward physics to apply force to substrate-attached cells. In this method, collagen-coated magnetic ferric oxide beads attach to the dorsal surface of cells via receptors of the integrin family, and a magnetic field gradient is applied to produce a force. In this paper we present a complete characterization of the method in a configuration that is easy to use, in which a permanent magnet provides a fairly uniform gradient over a relatively large area. This allows a fairly uniform average force that can be controlled in magnitude, direction, and duration to be applied to a large number of cells. We show how to determine the applied force per cell by measuring the force per unit volume of magnetic bead, the distribution of bead diameters, and the distribution of beads per cell. We also show how to calculate the force per unit volume of bead in a three-dimensional region near the permanent magnet on the basis of field measurements, and present results for three of the magnets. An upward force applied to fibroblasts by this method produces a measurable time-dependent increase in attachment of cytoskeletal actin filaments to the force application points, and an increase in actin cross-linking. This is accompanied by an actin-dependent retraction of the force-induced upward movement of the dorsal surface of the cells.
Glogauer
M
M
MRC Group in Periodontal Physiology, University of Toronto, Toronto, Ontario, Canada M5S 1A8.
Ferrier
J
J
D000199:Actins / Q000378:metabolism; D002477:Cells / Q000648:ultrastructure; D003599:Cytoskeleton / Q000737:chemistry; D005290:Ferric Compounds; D016023:Integrins; D008280:Magnetics; D008863:Microspheres; D013314:Stress, Mechanical
D000199:Actins; D005290:Ferric Compounds; D016023:Integrins; C000499:ferric oxide