TY - JOUR
T1 - Feedback-tracking microrheology in living cells
AU - Nishizawa, Kenji
AU - Bremerich, Marcel
AU - Ayade, Heev
AU - Schmidt, Christoph F.
AU - Ariga, Takayuki
AU - Mizuno, Daisuke
N1 - Funding Information:
Funding: This work was supported by the Japan Society for the Promotion of Science KAKENHI (grant numbers JP15H01494, JP25127712, JP25103011, JP15H03710, and JP23684036 to D.M.; grant numbers JP25870173 and JP15K05248 to T.A.; and grant number JP15J04464 to K.N.). K.N. was also supported by the Sasakawa Scientific Research Grant from the Japan Science Society (grant number 26–219). C.F.S. was supported by the Deutsche Forschungsgemeinschaft Collaborative Research Center SFB 937 (Project A2) and the European Research Council Advanced Grant FP7 ERC-2013-AdG, Project 324 340528. Author contributions: K.N. and M.B. conducted experiments and analysis. H.A. provided an analysis script. D.M. designed the research. D.M., K.N., M.B., T.A., and C.F.S. discussed the results and wrote the manuscript. Competing interests: The authors declare that they have no competing interests. Data and materials availability: All data needed to evaluate the conclusions in the paper are present in the paper and/or the Supplementary Materials. Additional data related to this paper may be requested from the authors
Publisher Copyright:
Copyright © 2017 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science.
PY - 2017
Y1 - 2017
N2 - Living cells are composed of active materials, in which forces are generated by the energy derived from metabolism. Forces and structures self-organize to shape the cell and drive its dynamic functions. Understanding the out-of-equilibrium mechanics is challenging because constituent materials, the cytoskeleton and the cytosol, are extraordinarily heterogeneous, and their physical properties are strongly affected by the internally generated forces. We have analyzed dynamics inside two types of eukaryotic cells, fibroblasts and epithelial-like HeLa cells, with simultaneous active and passive microrheology using laser interferometry and optical trapping technology. We developed a method to track microscopic probes stably in cells in the presence of vigorous cytoplasmic fluctuations, by using smooth three-dimensional (3D) feedback of a piezo-actuated sample stage. To interpret the data, we present a theory that adapts the fluctuation-dissipation theorem (FDT) to out-of-equilibrium systems that are subjected to positional feedback, which introduces an additional nonequilibrium effect. We discuss the interplay between material properties and nonthermal force fluctuations in the living cells that we quantify through the violations of the FDT. In adherent fibroblasts, we observed a well-known polymer network viscoelastic response where the complex shear modulus scales as G* º(−iw)3/4. In the more 3D confluent epithelial cells, we found glassy mechanics with G* º(−iw)1/2 that we attribute to glassy dynamics in the cytosol. The glassy state in living cells shows characteristics that appear distinct from classical glasses and unique to nonequilibrium materials that are activated by molecular motors.
AB - Living cells are composed of active materials, in which forces are generated by the energy derived from metabolism. Forces and structures self-organize to shape the cell and drive its dynamic functions. Understanding the out-of-equilibrium mechanics is challenging because constituent materials, the cytoskeleton and the cytosol, are extraordinarily heterogeneous, and their physical properties are strongly affected by the internally generated forces. We have analyzed dynamics inside two types of eukaryotic cells, fibroblasts and epithelial-like HeLa cells, with simultaneous active and passive microrheology using laser interferometry and optical trapping technology. We developed a method to track microscopic probes stably in cells in the presence of vigorous cytoplasmic fluctuations, by using smooth three-dimensional (3D) feedback of a piezo-actuated sample stage. To interpret the data, we present a theory that adapts the fluctuation-dissipation theorem (FDT) to out-of-equilibrium systems that are subjected to positional feedback, which introduces an additional nonequilibrium effect. We discuss the interplay between material properties and nonthermal force fluctuations in the living cells that we quantify through the violations of the FDT. In adherent fibroblasts, we observed a well-known polymer network viscoelastic response where the complex shear modulus scales as G* º(−iw)3/4. In the more 3D confluent epithelial cells, we found glassy mechanics with G* º(−iw)1/2 that we attribute to glassy dynamics in the cytosol. The glassy state in living cells shows characteristics that appear distinct from classical glasses and unique to nonequilibrium materials that are activated by molecular motors.
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U2 - 10.1126/sciadv.1700318
DO - 10.1126/sciadv.1700318
M3 - Article
C2 - 28975148
AN - SCOPUS:85033703639
VL - 3
JO - Science advances
JF - Science advances
SN - 2375-2548
IS - 9
M1 - 1700318
ER -