TY - JOUR
T1 - Probing the stochastic, motor-driven properties of the cytoplasm using force spectrum microscopy
AU - Guo, Ming
AU - Ehrlicher, Allen J.
AU - Jensen, Mikkel H.
AU - Renz, Malte
AU - Moore, Jeffrey R.
AU - Goldman, Robert D.
AU - Lippincott-Schwartz, Jennifer
AU - Mackintosh, Frederick C.
AU - Weitz, David A.
N1 - Funding Information:
We thank A. Rowat, S. Lindström, K. Kasza for helpful discussions; we thank F. Deng and A. Pegoraro for comments on the manuscript. This work was supported by the NIH (PO1GM096971), the Harvard Materials Research Science and Engineering Center (DMR-0820484), the NSF (DMR-1310266). A.J.E. was supported by NIH grant DK083592; M.H.J. and J.R.M. were supported by NIH grants HL86655 and HL077280; F.C.M. was supported in part by FOM/NWO. R.D.G. was supported by NIH PO1GM096971 and Hannah’s Hope Fund.
PY - 2014/8/14
Y1 - 2014/8/14
N2 - Molecular motors in cells typically produce highly directed motion; however, the aggregate, incoherent effect of all active processes also creates randomly fluctuating forces, which drive diffusive-like, nonthermal motion. Here, we introduce force-spectrum-microscopy (FSM) to directly quantify random forces within the cytoplasm of cells and thereby probe stochastic motor activity. This technique combines measurements of the random motion of probe particles with independent micromechanical measurements of the cytoplasm to quantify the spectrum of force fluctuations. Using FSM, we show that force fluctuations substantially enhance intracellular movement of small and large components. The fluctuations are three times larger in malignant cells than in their benign counterparts. We further demonstrate that vimentin acts globally to anchor organelles against randomly fluctuating forces in the cytoplasm, with no effect on their magnitude. Thus, FSM has broad applications for understanding the cytoplasm and its intracellular processes in relation to cell physiology in healthy and diseased states.
AB - Molecular motors in cells typically produce highly directed motion; however, the aggregate, incoherent effect of all active processes also creates randomly fluctuating forces, which drive diffusive-like, nonthermal motion. Here, we introduce force-spectrum-microscopy (FSM) to directly quantify random forces within the cytoplasm of cells and thereby probe stochastic motor activity. This technique combines measurements of the random motion of probe particles with independent micromechanical measurements of the cytoplasm to quantify the spectrum of force fluctuations. Using FSM, we show that force fluctuations substantially enhance intracellular movement of small and large components. The fluctuations are three times larger in malignant cells than in their benign counterparts. We further demonstrate that vimentin acts globally to anchor organelles against randomly fluctuating forces in the cytoplasm, with no effect on their magnitude. Thus, FSM has broad applications for understanding the cytoplasm and its intracellular processes in relation to cell physiology in healthy and diseased states.
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U2 - 10.1016/j.cell.2014.06.051
DO - 10.1016/j.cell.2014.06.051
M3 - Article
C2 - 25126787
AN - SCOPUS:84907369302
VL - 158
SP - 822
EP - 832
JO - Cell
JF - Cell
SN - 0092-8674
IS - 4
ER -