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. 2002 Dec;83(6):3162–3176. doi: 10.1016/S0006-3495(02)75319-8

Micromechanical mapping of live cells by multiple-particle-tracking microrheology.

Yiider Tseng 1, Thomas P Kole 1, Denis Wirtz 1
PMCID: PMC1302394  PMID: 12496086

Abstract

This paper introduces the method of live-cell multiple-particle-tracking microrheology (MPTM), which quantifies the local mechanical properties of living cells by monitoring the Brownian motion of individual microinjected fluorescent particles. Particle tracking of carboxylated microspheres imbedded in the cytoplasm produce spatial distributions of cytoplasmic compliances and frequency-dependent viscoelastic moduli. Swiss 3T3 fibroblasts are found to behave like a stiff elastic material when subjected to high rates of deformations and like a soft liquid at low rates of deformations. By analyzing the relative contributions of the subcellular compliances to the mean compliance, we find that the cytoplasm is much more mechanically heterogeneous than reconstituted actin filament networks. Carboxylated microspheres embedded in cytoplasm through endocytosis and amine-modified polystyrene microspheres, which are microinjected or endocytosed, often show directed motion and strong nonspecific interactions with cytoplasmic proteins, which prevents computation of local moduli from the microsphere displacements. Using MPTM, we investigate the mechanical function of alpha-actinin in non-muscle cells: alpha-actinin-microinjected cells are stiffer and yet mechanically more heterogeneous than control cells, in agreement with models of reconstituted cross-linked actin filament networks. MPTM is a new type of functional microscopy that can test the local, rate-dependent mechanical and ultrastructural properties of living cells.

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Selected References

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  1. Apgar J., Tseng Y., Fedorov E., Herwig M. B., Almo S. C., Wirtz D. Multiple-particle tracking measurements of heterogeneities in solutions of actin filaments and actin bundles. Biophys J. 2000 Aug;79(2):1095–1106. doi: 10.1016/S0006-3495(00)76363-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baschong W., Duerrenberger M., Mandinova A., Suetterlin R. Three-dimensional visualization of cytoskeleton by confocal laser scanning microscopy. Methods Enzymol. 1999;307:173–189. doi: 10.1016/s0076-6879(99)07013-5. [DOI] [PubMed] [Google Scholar]
  3. Bausch A. R., Möller W., Sackmann E. Measurement of local viscoelasticity and forces in living cells by magnetic tweezers. Biophys J. 1999 Jan;76(1 Pt 1):573–579. doi: 10.1016/S0006-3495(99)77225-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Beckerle M. C. Microinjected fluorescent polystyrene beads exhibit saltatory motion in tissue culture cells. J Cell Biol. 1984 Jun;98(6):2126–2132. doi: 10.1083/jcb.98.6.2126. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bray D., White J. G. Cortical flow in animal cells. Science. 1988 Feb 19;239(4842):883–888. doi: 10.1126/science.3277283. [DOI] [PubMed] [Google Scholar]
  6. Burridge K., Chrzanowska-Wodnicka M. Focal adhesions, contractility, and signaling. Annu Rev Cell Dev Biol. 1996;12:463–518. doi: 10.1146/annurev.cellbio.12.1.463. [DOI] [PubMed] [Google Scholar]
  7. Coulombe P. A., Bousquet O., Ma L., Yamada S., Wirtz D. The 'ins' and 'outs' of intermediate filament organization. Trends Cell Biol. 2000 Oct;10(10):420–428. doi: 10.1016/s0962-8924(00)01828-6. [DOI] [PubMed] [Google Scholar]
  8. Davies P. F., Barbee K. A., Volin M. V., Robotewskyj A., Chen J., Joseph L., Griem M. L., Wernick M. N., Jacobs E., Polacek D. C. Spatial relationships in early signaling events of flow-mediated endothelial mechanotransduction. Annu Rev Physiol. 1997;59:527–549. doi: 10.1146/annurev.physiol.59.1.527. [DOI] [PubMed] [Google Scholar]
  9. Domke J., Parak W. J., George M., Gaub H. E., Radmacher M. Mapping the mechanical pulse of single cardiomyocytes with the atomic force microscope. Eur Biophys J. 1999;28(3):179–186. doi: 10.1007/s002490050198. [DOI] [PubMed] [Google Scholar]
  10. Goodman Alan, Tseng Yiider, Wirtz Denis. Effect of length, topology, and concentration on the microviscosity and microheterogeneity of DNA solutions. J Mol Biol. 2002 Oct 18;323(2):199–215. doi: 10.1016/s0022-2836(02)00893-8. [DOI] [PubMed] [Google Scholar]
  11. Grazi E., Cuneo P., Magri E., Schwienbacher C., Trombetta G. Diffusion hindrance and geometry of filament crossings account for the complex interactions of F-actin with alpha-actinin from chicken gizzard. Biochemistry. 1993 Aug 31;32(34):8896–8901. doi: 10.1021/bi00085a022. [DOI] [PubMed] [Google Scholar]
  12. Haber C., Ruiz S. A., Wirtz D. Shape anisotropy of a single random-walk polymer. Proc Natl Acad Sci U S A. 2000 Sep 26;97(20):10792–10795. doi: 10.1073/pnas.190320097. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hall A. Rho GTPases and the actin cytoskeleton. Science. 1998 Jan 23;279(5350):509–514. doi: 10.1126/science.279.5350.509. [DOI] [PubMed] [Google Scholar]
  14. Hoh J. H., Schoenenberger C. A. Surface morphology and mechanical properties of MDCK monolayers by atomic force microscopy. J Cell Sci. 1994 May;107(Pt 5):1105–1114. doi: 10.1242/jcs.107.5.1105. [DOI] [PubMed] [Google Scholar]
  15. Izaguirre G., Aguirre L., Hu Y. P., Lee H. Y., Schlaepfer D. D., Aneskievich B. J., Haimovich B. The cytoskeletal/non-muscle isoform of alpha-actinin is phosphorylated on its actin-binding domain by the focal adhesion kinase. J Biol Chem. 2001 May 21;276(31):28676–28685. doi: 10.1074/jbc.M101678200. [DOI] [PubMed] [Google Scholar]
  16. Jarnik M., Aebi U. Toward a more complete 3-D structure of the nuclear pore complex. J Struct Biol. 1991 Dec;107(3):291–308. doi: 10.1016/1047-8477(91)90054-z. [DOI] [PubMed] [Google Scholar]
  17. LeDuc P., Haber C., Bao G., Wirtz D. Dynamics of individual flexible polymers in a shear flow. Nature. 1999 Jun 10;399(6736):564–566. doi: 10.1038/21148. [DOI] [PubMed] [Google Scholar]
  18. Lo C. M., Wang H. B., Dembo M., Wang Y. L. Cell movement is guided by the rigidity of the substrate. Biophys J. 2000 Jul;79(1):144–152. doi: 10.1016/S0006-3495(00)76279-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Ma L., Yamada S., Wirtz D., Coulombe P. A. A 'hot-spot' mutation alters the mechanical properties of keratin filament networks. Nat Cell Biol. 2001 May;3(5):503–506. doi: 10.1038/35074576. [DOI] [PubMed] [Google Scholar]
  20. Matzke R., Jacobson K., Radmacher M. Direct, high-resolution measurement of furrow stiffening during division of adherent cells. Nat Cell Biol. 2001 Jun;3(6):607–610. doi: 10.1038/35078583. [DOI] [PubMed] [Google Scholar]
  21. Palmer A., Xu J., Kuo S. C., Wirtz D. Diffusing wave spectroscopy microrheology of actin filament networks. Biophys J. 1999 Feb;76(2):1063–1071. doi: 10.1016/S0006-3495(99)77271-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Parent C. A., Devreotes P. N. A cell's sense of direction. Science. 1999 Apr 30;284(5415):765–770. doi: 10.1126/science.284.5415.765. [DOI] [PubMed] [Google Scholar]
  23. Pavalko F. M., Burridge K. Disruption of the actin cytoskeleton after microinjection of proteolytic fragments of alpha-actinin. J Cell Biol. 1991 Aug;114(3):481–491. doi: 10.1083/jcb.114.3.481. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Penman S. Rethinking cell structure. Proc Natl Acad Sci U S A. 1995 Jun 6;92(12):5251–5257. doi: 10.1073/pnas.92.12.5251. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Qian H., Sheetz M. P., Elson E. L. Single particle tracking. Analysis of diffusion and flow in two-dimensional systems. Biophys J. 1991 Oct;60(4):910–921. doi: 10.1016/S0006-3495(91)82125-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Radmacher M., Cleveland J. P., Fritz M., Hansma H. G., Hansma P. K. Mapping interaction forces with the atomic force microscope. Biophys J. 1994 Jun;66(6):2159–2165. doi: 10.1016/S0006-3495(94)81011-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Ragsdale G. K., Phelps J., Luby-Phelps K. Viscoelastic response of fibroblasts to tension transmitted through adherens junctions. Biophys J. 1997 Nov;73(5):2798–2808. doi: 10.1016/S0006-3495(97)78309-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Rivero F., Furukawa R., Fechheimer M., Noegel A. A. Three actin cross-linking proteins, the 34 kDa actin-bundling protein, alpha-actinin and gelation factor (ABP-120), have both unique and redundant roles in the growth and development of Dictyostelium. J Cell Sci. 1999 Aug;112(Pt 16):2737–2751. doi: 10.1242/jcs.112.16.2737. [DOI] [PubMed] [Google Scholar]
  29. Rotsch C., Braet F., Wisse E., Radmacher M. AFM imaging and elasticity measurements on living rat liver macrophages. Cell Biol Int. 1997 Nov;21(11):685–696. doi: 10.1006/cbir.1997.0213. [DOI] [PubMed] [Google Scholar]
  30. Sato M., Schwarz W. H., Pollard T. D. Dependence of the mechanical properties of actin/alpha-actinin gels on deformation rate. 1987 Feb 26-Mar 4Nature. 325(6107):828–830. doi: 10.1038/325828a0. [DOI] [PubMed] [Google Scholar]
  31. Saxton M. J. Anomalous diffusion due to obstacles: a Monte Carlo study. Biophys J. 1994 Feb;66(2 Pt 1):394–401. doi: 10.1016/s0006-3495(94)80789-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Schoenenberger C. A., Steinmetz M. O., Stoffler D., Mandinova A., Aebi U. Structure, assembly, and dynamics of actin filaments in situ and in vitro. Microsc Res Tech. 1999 Oct 1;47(1):38–50. doi: 10.1002/(SICI)1097-0029(19991001)47:1<38::AID-JEMT4>3.0.CO;2-5. [DOI] [PubMed] [Google Scholar]
  33. Svitkina T. M., Borisy G. G. Correlative light and electron microscopy of the cytoskeleton of cultured cells. Methods Enzymol. 1998;298:570–592. doi: 10.1016/s0076-6879(98)98045-4. [DOI] [PubMed] [Google Scholar]
  34. Tempel M, Isenberg G, Sackmann E. Temperature-induced sol-gel transition and microgel formation in alpha -actinin cross-linked actin networks: A rheological study. Phys Rev E Stat Phys Plasmas Fluids Relat Interdiscip Topics. 1996 Aug;54(2):1802–1810. doi: 10.1103/physreve.54.1802. [DOI] [PubMed] [Google Scholar]
  35. Tseng Y., Fedorov E., McCaffery J. M., Almo S. C., Wirtz D. Micromechanics and ultrastructure of actin filament networks crosslinked by human fascin: a comparison with alpha-actinin. J Mol Biol. 2001 Jul 6;310(2):351–366. doi: 10.1006/jmbi.2001.4716. [DOI] [PubMed] [Google Scholar]
  36. Tseng Y., Wirtz D. Mechanics and multiple-particle tracking microheterogeneity of alpha-actinin-cross-linked actin filament networks. Biophys J. 2001 Sep;81(3):1643–1656. doi: 10.1016/S0006-3495(01)75818-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Vassy J., Irinopoulou T., Beil M., Rigaut J. P. Spatial distribution of cytoskeleton intermediate filaments during fetal rat hepatocyte differentiation. Microsc Res Tech. 1997 Dec 1;39(5):436–443. doi: 10.1002/(SICI)1097-0029(19971201)39:5<436::AID-JEMT6>3.0.CO;2-E. [DOI] [PubMed] [Google Scholar]
  38. Wachsstock D. H., Schwartz W. H., Pollard T. D. Affinity of alpha-actinin for actin determines the structure and mechanical properties of actin filament gels. Biophys J. 1993 Jul;65(1):205–214. doi: 10.1016/S0006-3495(93)81059-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Wachsstock D. H., Schwarz W. H., Pollard T. D. Cross-linker dynamics determine the mechanical properties of actin gels. Biophys J. 1994 Mar;66(3 Pt 1):801–809. doi: 10.1016/s0006-3495(94)80856-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Wang N., Ingber D. E. Probing transmembrane mechanical coupling and cytomechanics using magnetic twisting cytometry. Biochem Cell Biol. 1995 Jul-Aug;73(7-8):327–335. doi: 10.1139/o95-041. [DOI] [PubMed] [Google Scholar]
  41. Xu J., Tseng Y., Wirtz D. Strain hardening of actin filament networks. Regulation by the dynamic cross-linking protein alpha-actinin. J Biol Chem. 2000 Nov 17;275(46):35886–35892. doi: 10.1074/jbc.M002377200. [DOI] [PubMed] [Google Scholar]
  42. Xu J., Wirtz D., Pollard T. D. Dynamic cross-linking by alpha-actinin determines the mechanical properties of actin filament networks. J Biol Chem. 1998 Apr 17;273(16):9570–9576. doi: 10.1074/jbc.273.16.9570. [DOI] [PubMed] [Google Scholar]
  43. Xu Jingyuan, Tseng Yiider, Carriere Craig J., Wirtz Denis. Microheterogeneity and microrheology of wheat gliadin suspensions studied by multiple-particle tracking. Biomacromolecules. 2002 Jan-Feb;3(1):92–99. doi: 10.1021/bm015586b. [DOI] [PubMed] [Google Scholar]
  44. Yamada S., Wirtz D., Kuo S. C. Mechanics of living cells measured by laser tracking microrheology. Biophys J. 2000 Apr;78(4):1736–1747. doi: 10.1016/S0006-3495(00)76725-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Yamashiro S., Yamakita Y., Ono S., Matsumura F. Fascin, an actin-bundling protein, induces membrane protrusions and increases cell motility of epithelial cells. Mol Biol Cell. 1998 May;9(5):993–1006. doi: 10.1091/mbc.9.5.993. [DOI] [PMC free article] [PubMed] [Google Scholar]

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