Physical Properties of the Nucleus Studied by Micropipette Aspiration

  • Amy C. Rowat
Part of the Methods in Molecular Biology book series (MIMB, volume 464)


Understanding the physical properties of the cell nucleus is critical for developing a deeper understanding of nuclear structure and organization as well as how mechanical forces induce changes in gene expression. We use micropipette aspiration to induce large, local deformations in the nucleus, and microscopy to image nuclear shape as well as the response of fluorescently labeled components in the inner nucleus (chromatin and nucleoli) and the nuclear envelope (lamins and membranes). By monitoring the response of nuclear structures to these deformations, we gain insights into the material properties of the nucleus. Here we describe the experimental protocols for micropipette aspiration of nuclei in living cells as well as isolated nuclei. In addition to confocal imaging, deformed nuclei can be imaged by brightfield or epifluorescence microscopy.


Nuclear mechanics Micropipette aspiration Confocal imaged microdeformation 



Many thanks to J.H. Ipsen for critical discussions. Thanks also to D.K. Shumaker and R.D. Goldman, Northwestern University, Chicago, USA for generously providing the GFP-Lam A construct as well as to J.S. Andersen, Y.W. Lam, and J. Lammerding for helpful advice. This work was supported by the Danish National Research Foundation and a NSERC Julie Payette Scholarship (ACR). ACR is a Human Frontiers Science Program Cross-disciplinary fellow.


  1. 1.
    Garcia-Cardena, G., Comander, J., Anderson. K.R., Blackman, B.R., and Gimbrone, M.A. Jr. (2001) Biomechanical activation of vascular endothelium as a determinant of its functional phenotype. Proc. Natl. Acad. Sci. USA 98, 4478-4485PubMedCrossRefGoogle Scholar
  2. 2.
    Mattout, A., Dechat, T., Adam, S.A., Goldman, R.D., and Gruenbaum, Y. (2006) Nuclear lamins, diseases and aging. Curr. Opin. Cell Biol. 18, 335-341PubMedCrossRefGoogle Scholar
  3. 3.
    Broers, J.L, Ramaekers, F.C., Bonne, G., Yaou, R.B., and Hutchison, C.J. (2006) Nuclear lamins: laminopathies and their role in premature ageing. Physiol. Rev. 86, 967-1008PubMedCrossRefGoogle Scholar
  4. 4.
    Rowat, A.C., Lammerding, J., Herrmann, H., and Aebi, U. (2008) Towards an integrated understanding of the structure and mechanics of the cell nucleus. Bioessays 30, 226-236PubMedCrossRefGoogle Scholar
  5. 5.
    Dahl, K.N., Engler, A.J., Pajerowski, J.D., and Discher, D.E. (2005) Power-law rheology of isolated nuclei with deformation mapping of nuclear substructures. Biophys. J. 89, 2855-2864PubMedCrossRefGoogle Scholar
  6. 6.
    Guilak, F., Tedrow, J.R., and Burgkart, R. (2000) Viscoelastic properties of the cell nucleus. Biochem. Biophys. Res. Commun. 269, 781-786PubMedCrossRefGoogle Scholar
  7. 7.
    Rowat, A.C., Foster, L.J., Nielsen, M.M., Weiss, M., and Ipsen, J.H. (2005) Characterization of the elastic properties of the nuclear envelope. J. R. Soc. Interface 2, 63-69PubMedCrossRefGoogle Scholar
  8. 8.
    Rowat, A.C., Lammerding, J., and Ipsen, J.H. (2006) Mechanical properties of the cell nucleus and the effect of emerin deficiency. Biophys. J. 91, 1-16CrossRefGoogle Scholar
  9. 9.
    Moir, R.D., Yoon, M., Khuon, S., and Goldman, RMD. (2000) Nuclear lamins A and B1: dif-ferent pathways of assembly during nuclear envelope formation in living cells. J. Cell Biol. 151, 1155-1168PubMedCrossRefGoogle Scholar
  10. 10.
    Jackson, D.A., Yuan, J., and Cook, P.R. (1988) A gentle method for preparing cyto- and nucleo-skeletons and associated chromatin. J. Cell Sci. 90, 365-378PubMedGoogle Scholar
  11. 11.
    Henriksen, J.R. and Ipsen, J.H. (2004) Measurement of membrane elasticity by micro-pipette aspiration. Eur. Phys. J. E Soft Matter 14, 149-167PubMedCrossRefGoogle Scholar
  12. 12.
    Evans, E. and Needham, D. (1987) Physical properties of surfactant bilayer membranes: thermal transitions, elasticity, rigidity, cohesion, and colloidal interactions. J. Phys. Chem. 91, 4219-4228CrossRefGoogle Scholar
  13. 13.
    Bojanowski, K. and Ingber, D.E. (1998) Ionic control of chromosome architecture in living and permeabilized cells. Exp. Cell Res. 244, 286-294PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press, a part of Springer Science + Business Media, LLC 2008

Authors and Affiliations

  • Amy C. Rowat
    • 1
  1. 1.Department of Physics/Division of Engineering and Applied ScienceHarvard University, Engineering Sciences LaboratoryCambridgeUSA

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