Nanoscopy of bacterial cells immobilized by holographic optical tweezers

Robin Diekmann, Deanna L. Wolfson, Christoph Spahn, Mike Heilemann, Mark Schüttpelz and Thomas Huser ()
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Robin Diekmann: Biomolecular Photonics, University of Bielefeld
Deanna L. Wolfson: NSF Center for Biophotonics, University of California
Christoph Spahn: Institute of Physical and Theoretical Chemistry, Johann Wolfgang Goethe-University Frankfurt
Mike Heilemann: Institute of Physical and Theoretical Chemistry, Johann Wolfgang Goethe-University Frankfurt
Mark Schüttpelz: Biomolecular Photonics, University of Bielefeld
Thomas Huser: Biomolecular Photonics, University of Bielefeld

Nature Communications, 2016, vol. 7, issue 1, 1-7

Abstract: Abstract Imaging non-adherent cells by super-resolution far-field fluorescence microscopy is currently not possible because of their rapid movement while in suspension. Holographic optical tweezers (HOTs) enable the ability to freely control the number and position of optical traps, thus facilitating the unrestricted manipulation of cells in a volume around the focal plane. Here we show that immobilizing non-adherent cells by optical tweezers is sufficient to achieve optical resolution well below the diffraction limit using localization microscopy. Individual cells can be oriented arbitrarily but preferably either horizontally or vertically relative to the microscope’s image plane, enabling access to sample sections that are impossible to achieve with conventional sample preparation and immobilization. This opens up new opportunities to super-resolve the nanoscale organization of chromosomal DNA in individual bacterial cells.

Date: 2016
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DOI: 10.1038/ncomms13711

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