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Scanning Near-Field Optical Microscopy and Microspectroscopy of Green Fluorescent Protein in Intact Escherichia coli Bacteria

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Abstract

Scanning near-field optical microscopy (SNOM) yields high-resolution topographic and optical information and constitutes an important new technique for visualizing biological systems. By coupling a spectrograph to a near-field microscope, we have been able to perform microspectroscopic measurements with a spatial resolution greatly exceeding that of the conventional optical microscope. Here we present SNOM images of Escherichia coli bacteria expressing a mutant green fluorescent protein (GFP), an important reporter molecule in cell, developmental, and molecular biology. Near-field emission spectra confirm that the fluorescence detected by SNOM arises from bacterially expressed GFP molecules.

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Authors and Affiliations

  1. Department of Molecular Biology, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, D-37077, Göttingen, Germany

    Vinod Subramaniam, Achim K. Kirsch & Thomas M. Jovin

  2. Department of Molecular Developmental Biology, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, D-37077, Göttingen, Germany

    Rolando V. Rivera-Pomar

Authors
  1. Vinod Subramaniam
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  2. Achim K. Kirsch
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  3. Rolando V. Rivera-Pomar
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  4. Thomas M. Jovin
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Correspondence to Thomas M. Jovin.

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Subramaniam, V., Kirsch, A.K., Rivera-Pomar, R.V. et al. Scanning Near-Field Optical Microscopy and Microspectroscopy of Green Fluorescent Protein in Intact Escherichia coli Bacteria. Journal of Fluorescence 7, 381–385 (1997). https://doi.org/10.1023/A:1022554715488

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