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Force fluctuations in three-dimensional suspended fibroblasts.

Philosophical transactions of the Royal Society of London. Series B, Biological sciences (2014-12-24)
Florian Schlosser, Florian Rehfeldt, Christoph F Schmidt
ABSTRACT

Cells are sensitive to mechanical cues from their environment and at the same time generate and transmit forces to their surroundings. To test quantitatively forces generated by cells not attached to a substrate, we used a dual optical trap to suspend 3T3 fibroblasts between two fibronectin-coated beads. In this simple geometry, we measured both the cells' elastic properties and the force fluctuations they generate with high bandwidth. Cell stiffness decreased substantially with both myosin inhibition by blebbistatin and serum-starvation, but not with microtubule depolymerization by nocodazole. We show that cortical forces generated by non-muscle myosin II deform the cell from its rounded shape in the frequency regime from 0.1 to 10 Hz. The amplitudes of these forces were strongly reduced by blebbistatin and serum starvation, but were unaffected by depolymerization of microtubules. Force fluctuations show a spectrum that is characteristic for an elastic network activated by random sustained stresses with abrupt transitions.

MATERIALS
Product Number
Brand
Product Description

Avanti
18:1 Biotinyl Cap PE, Avanti Research - A Croda Brand 870273C
Avanti
18:1 Biotinyl Cap PE, Avanti Research - A Croda Brand 870273P, powder
Avanti
18:1 (Δ9-Cis) PC (DOPC), Avanti Research - A Croda Brand
Avanti
18:1 (Δ9-Cis) PC (DOPC), Avanti Research - A Croda Brand