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Programmed subcellular release for studying the dynamics of cell detachment

Nature Methods volume 6, pages 211–213 (2009)Cite this article

Abstract

Cell detachment is central to a broad range of physiopathological changes, but there are no quantitative methods to study this process. Here we report programmed subcellular release, a method for spatially and temporally controlled cellular detachment, and present quantitative results of the detachment dynamics of 3T3 fibroblasts at the subcellular level.

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Figure 1: Programmed subcellular release.
Figure 2: Programmed subcellular release in the presence of cytoskeleton inhibitors.
Figure 3: Cell detachment and focal adhesion dynamics.

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Acknowledgements

This work was supported by US National Institutes of Health (D.W. and P.C.S.), National Science Foundation (P.C.S.), Howard Hughes Medical Institution (D.W. and P.C.S.) and the Achievement Awards for College Scientists foundation (B.W.). We thank members of Wirtz and Searson laboratories for technical advice and reagents, S. Sun for useful discussions and H. Vo for assistance with microfabrication of devices.

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Authors and Affiliations

  1. Department of Materials Science and Engineering, Johns Hopkins University, 3400 N. Charles St., Baltimore, 21218, Maryland, USA

    Bridget Wildt, Denis Wirtz & Peter C Searson

  2. Department of Chemical and Biomolecular Engineering, Johns Hopkins University, 3400 N. Charles St., Baltimore, 21218, Maryland, USA

    Denis Wirtz & Peter C Searson

  3. Howard Hughes Medical Institute Graduate Training Program and the Institute for NanoBioTechnology, Johns Hopkins University, 3400 N. Charles St., Baltimore, 21218, Maryland, USA

    Denis Wirtz & Peter C Searson

Authors
  1. Bridget Wildt
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  2. Denis Wirtz
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  3. Peter C Searson
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Corresponding authors

Correspondence to Denis Wirtz or Peter C Searson.

Supplementary information

Supplementary Text and Figure

Supplementary Figures 1–5 and Supplementary Methods (PDF 11186 kb)

Supplementary Movie 1

Time lapse movie of sub-cellular release (see Fig. 1f). Magnification, 10×. (MOV 12174 kb)

Supplementary Movie 2

Time lapse movie of sub-cellular release (see Supplementary Fig. 2a). Magnification, 10×. (MOV 7983 kb)

Supplementary Movie 3

Time lapse movie of sub-cellular release (see Supplementary Fig. 2b). Magnification, 60×. (MOV 18930 kb)

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Wildt, B., Wirtz, D. & Searson, P. Programmed subcellular release for studying the dynamics of cell detachment. Nat Methods 6, 211–213 (2009). https://doi.org/10.1038/nmeth.1299

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