Analysis of subcellular surface structure, function and dynamics |
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Authors: | D Anselmetti N Hansmeier J Kalinowski J Martini T Merkle R Palmisano R Ros K Schmied A Sischka K Toensing |
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Institution: | 1. Experimental Biophysics and Applied Nanoscience, Faculty of Physics, Bielefeld University, Universit?tsstra?e 25, 33615, Bielefeld, Germany 2. Institute for Genome Research, Center of Biotechnology, Bielefeld University, Universit?tsstra?e 25, 33615, Bielefeld, Germany
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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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Keywords: | Atomic force microscopy Optical tweezers Two-photon laser scanning microscopy |
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