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Molecular dynamics imaging in micropatterned living cells

Nature Methods volume 2, pages 739–741 (2005)Cite this article

Abstract

Micropatterning approaches using self-assembled monolayers of alkyl thiols on gold are not optimal for important imaging modalities in cell biology because of absorption of light and scattering of electrons by the gold layer. We report here an anisotropic solid microetching (ASOMIC) procedure that overcomes these limitations. The method allows molecular dynamics imaging by wide-field and total internal reflection fluorescence (TIRF) microscopy of living mammalian cells and correlative platinum replica electron microscopy.

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Figure 1: ASOMIC method allows for cell micropatterning compatible with fluorescence imaging and electron microscopy.
Figure 2: Microtubule edge impact turning depends on cell shape.

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Acknowledgements

This work was supported by US National Institutes of Health grant GM 25062 (G.G.B.), Camille and Henry Dreyfus New Faculty Award (B.A.G.), Baxter Health Care Corporation (G.G.B. and B.A.G.) and Department of Defense Breast Cancer Research Program postdoctoral fellowship (K.K.G.). We thank O. Chaga for help with electron microscopy.

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Authors and Affiliations

  1. Department of Cell and Molecular Biology, Northwestern University Medical School, 303 East Chicago Avenue, Chicago, 60611-3008, Illinois, USA

    Kristiana Kandere-Grzybowska, Yulia Komarova & Gary G Borisy

  2. Department of Chemical and Biological Engineering, Northwestern University, 2145 Sheridan Road, Evanston, 60208-3120, Illinois

    Christopher Campbell & Bartosz A Grzybowski

Authors
  1. Kristiana Kandere-Grzybowska
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  2. Christopher Campbell
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  3. Yulia Komarova
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  4. Bartosz A Grzybowski
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  5. Gary G Borisy
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Corresponding authors

Correspondence to Bartosz A Grzybowski or Gary G Borisy.

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Supplementary information

Supplementary Fig. 1

Distribution of focal adhesions, actin filaments and microtubules in micropatterned cells. (PDF 5595 kb)

Supplementary Fig. 2

Fluorescence signal attenuation by thin gold layers. (PDF 963 kb)

Supplementary Video 1

Microtubule dynamics in B16F1 mouse melanoma cells on un-patterned substrata. (MOV 2017 kb)

Supplementary Video 2

Microtubule dynamics in B16F1 cells on circular adhesive islands. (MOV 1618 kb)

Supplementary Video 3

Microtubule dynamics B16F1 mouse melanoma cells on triangular adhesive islands. (MOV 2190 kb)

Supplementary Methods (PDF 82 kb)

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Kandere-Grzybowska, K., Campbell, C., Komarova, Y. et al. Molecular dynamics imaging in micropatterned living cells. Nat Methods 2, 739–741 (2005). https://doi.org/10.1038/nmeth796

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