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  • Correspondence
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Reducing background fluorescence reveals adhesions in 3D matrices

Nature Cell Biology volume 13, pages 3–5 (2011)Cite this article

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An Addendum to this article was published on 30 November 2012

This article has been updated

To the editor:

Adhesion complexes in cells growing on planar substrates have been studied for over three decades. From these studies, several classes of adhesions have been described based on size, location, morphology, dynamics or molecular composition, and their dual role as signalling centres and linkages that connect the extracellular matrix (ECM) with the cytoskeleton has also emerged1. In contrast to this large and growing understanding of adhesion on two-dimensional (2D) substrates, little is known about the adhesions formed during three-dimensional (3D) growth of cells, including whether they even exist. Immunostaining and microscopy of fixed samples2,3,4,5,6,7,8 and dynamic imaging at low spatiotemporal resolution9 have shown the presence of large, elongated adhesions on cells in various 3D model systems. However, attempts to visualize adhesions in living cells growing in 3D with resolution similar to that routinely used in 2D have not been successful10. This has led to the recent conclusion that the adhesions defined and characterized in 2D cultures either do not exist in cells in 3D, or are too small or short-lived to be observed11.

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Figure 1: Cell-matrix adhesions are detected in 3D collagen gels.
Figure 2: Dynamics of cell-matrix adhesions in 3D culture.

Change history

  • 25 October 2012

    In the version of this Correspondence originally published, a manufacturing issue with the rat-tail collagen solution meant that in one Supplementary experiment (Fig. S1i–n) the collagen gel concentration was actually 1.2 mg ml-1 instead of the 2 mg ml-1 we reported. This does not change the conclusions of the paper; see PDF for further details.

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Acknowledgements

We thank C. Choi, M. Vicente-Manzanares, R. Tilghman, and E. Gratton for helpful discussions and technical assistance. The research was supported by the Cell Migration Consortium (U54 GM064346) and GM23244. K.E.K. is supported by a Cancer Training Grant at the University of Virginia Cancer Center (T32 CA009109-34).

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  1. Department of Cell Biology, University of Virginia, Charlottesville, 22908, VA, USA

    Kristopher E. Kubow & Alan Rick Horwitz

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  1. Kristopher E. Kubow
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K.E.K. performed the experiments and analysed the data. K.E.K. and A.R.H. designed the experiments and wrote the paper.

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Correspondence to Alan Rick Horwitz.

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The authors declare no competing financial interests.

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Kubow, K., Horwitz, A. Reducing background fluorescence reveals adhesions in 3D matrices. Nat Cell Biol 13, 3–5 (2011). https://doi.org/10.1038/ncb0111-3

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