Analysis of cell mechanics in single vinculin-deficient cells using a magnetic tweezer.
A magnetic tweezer was constructed to apply controlled tensional forces (10 pN to greater than 1 nN) to transmembrane receptors via bound ligand-coated microbeadswhile optically measuring lateral bead displacements within individual cells. Use of this system with wild-type F9 embryonic carcinoma cells and cells from a vinculin knockout mouse F9 Vin (-/-) revealed much larger differences in the stiffness of the transmembrane integrin linkages to the cytoskeleton than previously reported using related techniques that measured average mechanical properties of large cell populations. The mechanical properties measured varied widely among cells, exhibiting an approximately log-normal distribution. The median lateral bead displacement was 2-fold larger in F9 Vin (-/-) cells compared to wild-type cells whereas the arithmetic mean displacement only increased by 37%. We conclude that vinculin serves a greater mechanical role in cells than previously reported and that this magnetic tweezer device may be useful for probing the molecular basis of cell mechanics within single cells.
Alenghat
F J
FJ
Department of Pathology, Children's Hospital and Harvard Medical School, Boston, Massachusetts 02115, USA.
Fabry
B
B
Tsai
K Y
KY
Goldmann
W H
WH
Ingber
D E
DE
D000818:Animals; D002138:Calibration; D002462:Cell Membrane / Q000502:physiology*; D002478:Cells, Cultured; D003599:Cytoskeleton / Q000378:metabolism; D016023:Integrins / Q000378:metabolism; D008280:Magnetics; D051379:Mice; D018345:Mice, Knockout; D008863:Microspheres; D011956:Receptors, Cell Surface / Q000378:metabolism; D013314:Stress, Mechanical; D014407:Tumor Cells, Cultured; D016596:Vinculin / Q000172:deficiency* / Q000235:genetics / Q000502:physiology*
D016023:Integrins; D011956:Receptors, Cell Surface; D016596:Vinculin