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Physical Properties of the Nucleus Studied by Micropipette Aspiration

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

Abstract

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.

Keywords

Nuclear mechanics Micropipette aspiration Confocal imaged microdeformation 

Notes

Acknowledgments

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.

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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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