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Analysis of subcellular surface structure, function and dynamics

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Abstract

Analytics of single biological cells allows quantitative investigation from a structural, functional and dynamical point of view and opens novel possibilities to an unamplified subcellular analysis. In this article, we report on three different experimental methods and their applications to single cellular systems with a subcellular sensitivity down to the single molecule level. First, the subcellular surface structure of living bacteria (Corynebacterium glutamicum) was investigated with atomic force microscopy (AFM) at the resolution of individual surface layer (S-layer) proteins; discrimination of bacterial strains that lack the expression of hexagonally packed surface layer proteins was possible. Second, quantitative measurement of individual recognition events of membrane-bound receptors on living B-cells was achieved in single cell manipulation and probing experiments with optical tweezers (OT) force spectroscopy. And third, intracellular dynamics of translocating photoactivatable GFP in plant protoplasts (Nicotiana tabacum BY-2) was quantitatively monitored by two-photon laser scanning microscopy (2PLSM).

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Acknowledgements

Financial support from the Deutsche Forschungsgemeinschaft (DFG) within the Collaborative Research Project SFB 613-Physics of molecular recognition and single molecule processes in organic systems (D.A.), the Federal Ministry of Education and Research of Germany (BMBF) for the Research Project MEMO-Foundations of laser-based characterization of metabolism and morphology of biological tissue (D.A.) and from the DFG grants ME 1116/4–2 and BIZ 7/1–2 (T.M.) is gratefully acknowledged.

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Authors and Affiliations

  1. Experimental Biophysics and Applied Nanoscience, Faculty of Physics, Bielefeld University, Universitätsstraße 25, 33615, Bielefeld, Germany

    D. Anselmetti, J. Martini, R. Ros, A. Sischka & K. Toensing

  2. Institute for Genome Research, Center of Biotechnology, Bielefeld University, Universitätsstraße 25, 33615, Bielefeld, Germany

    N. Hansmeier, J. Kalinowski, T. Merkle, R. Palmisano & K. Schmied

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  1. D. Anselmetti
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  2. N. Hansmeier
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  3. J. Kalinowski
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  4. J. Martini
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  6. R. Palmisano
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  7. R. Ros
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  9. A. Sischka
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  10. K. Toensing
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Correspondence to D. Anselmetti.

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Anselmetti, D., Hansmeier, N., Kalinowski, J. et al. Analysis of subcellular surface structure, function and dynamics. Anal Bioanal Chem 387, 83–89 (2007). https://doi.org/10.1007/s00216-006-0789-3

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  • DOI: https://doi.org/10.1007/s00216-006-0789-3

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