Imaging of cell/substrate contacts of living cells with surface plasmon resonance microscopy

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243_biophysj_1999.pdf
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1999
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Giebel, Karl-Friedrich
Herminghaus, Stephan
Riedel, M.
Weiland, Ulrich
Bastmeyer, Martin
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Biophysical Journal. 1999, 76(1), pp. 509-516. Available under: doi: 10.1016/S0006-3495(99)77219-X
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We have developed a new method for observing cell/substrate contacts of living cells in culture based on the optical excitation of surface plasmons. Surface plasmons are quanta of an electromagnetic wave that travel along the interface between a metal and a dielectric layer. The evanescent field associated with this excitation decays exponentially perpendicular to the interface, on the order of some hundreds of nanometers. Cells were cultured on an aluminum-coated glass prism and illuminated from below with a laser beam. Because the cells interfere with the evanescent field, the intensity of the reflected light, which is projected onto a camera chip, correlates with the cell/substrate distance. Contacts between the cell membrane and the substrate can thus be visualized at high contrast with a vertical resolution in the nanometer range. The lateral resolution along the propagation direction of surface plasmons is given by their lateral momentum, whereas perpendicular to it, the resolution is determined by the optical diffraction limit. For quantitative analysis of cell/substrate distances, cells were imaged at various angles of incidence to obtain locally resolved resonance curves. By comparing our experimental data with theoretical surface plasmon curves we obtained a cell/substrate distance of 160 6 10 nm for most parts of the cells. Peripheral lamellipodia, in contrast, formed contacts with a cell substrate/distance of 25 6 10 nm.

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ISO 690GIEBEL, Karl-Friedrich, Clemens BECHINGER, Stephan HERMINGHAUS, M. RIEDEL, Paul LEIDERER, Ulrich WEILAND, Martin BASTMEYER, 1999. Imaging of cell/substrate contacts of living cells with surface plasmon resonance microscopy. In: Biophysical Journal. 1999, 76(1), pp. 509-516. Available under: doi: 10.1016/S0006-3495(99)77219-X
BibTex
@article{Giebel1999Imagi-4913,
  year={1999},
  doi={10.1016/S0006-3495(99)77219-X},
  title={Imaging of cell/substrate contacts of living cells with surface plasmon resonance microscopy},
  number={1},
  volume={76},
  journal={Biophysical Journal},
  pages={509--516},
  author={Giebel, Karl-Friedrich and Bechinger, Clemens and Herminghaus, Stephan and Riedel, M. and Leiderer, Paul and Weiland, Ulrich and Bastmeyer, Martin}
}
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