Get access

Three-Dimensional Exploration and Mechano-Biophysical Analysis of the Inner Structure of Living Cells

Authors

  • Álvaro Barroso,

    1. Institute of Applied Physics, University of Muenster, Corrensstrasse 2-4, 48149 Muenster, Germany
    Search for more papers by this author
  • Mike Woerdemann,

    1. Institute of Applied Physics, University of Muenster, Corrensstrasse 2-4, 48149 Muenster, Germany
    Search for more papers by this author
  • Angelika Vollmer,

    1. Center for Biomedical Optics and Photonics, University of Muenster, Robert-Koch-Strasse 45, 48149 Muenster, Germany
    Search for more papers by this author
  • Gert von Bally,

    1. Center for Biomedical Optics and Photonics, University of Muenster, Robert-Koch-Strasse 45, 48149 Muenster, Germany
    Search for more papers by this author
  • Björn Kemper,

    Corresponding author
    1. Center for Biomedical Optics and Photonics, University of Muenster, Robert-Koch-Strasse 45, 48149 Muenster, Germany
    • Björn Kemper, Center for Biomedical Optics and Photonics, University of Muenster, Robert-Koch-Strasse 45, 48149 Muenster, Germany.

      Cornelia Denz, Institute of Applied Physics, University of Muenster, Corrensstrasse 2-4, 48149 Muenster, Germany

    Search for more papers by this author
  • Cornelia Denz

    Corresponding author
    1. Institute of Applied Physics, University of Muenster, Corrensstrasse 2-4, 48149 Muenster, Germany
    • Björn Kemper, Center for Biomedical Optics and Photonics, University of Muenster, Robert-Koch-Strasse 45, 48149 Muenster, Germany.

      Cornelia Denz, Institute of Applied Physics, University of Muenster, Corrensstrasse 2-4, 48149 Muenster, Germany

    Search for more papers by this author

Abstract

A novel mechanobiological method is presented to explore qualitatively and quantitatively the inside of living biological cells in three dimensions, paving the way to sense intracellular changes during dynamic cellular processes. For this purpose, holographic optical tweezers, which allow the versatile manipulation of nanoscopic and microscopic particles by means of tailored light fields, are combined with self-interference digital holographic microscopy. This biophotonic holographic workstation enables non-contact, minimally invasive, flexible, high-precision optical manipulation and accurate 3D tracking of probe particles that are incorporated by phagocytosis in cells, while simultaneously quantitatively phase imaging the cell morphology. In a first model experiment, internalized polystyrene microspheres with 1 μm diameter are three-dimensionally moved and tracked in order to quantify distances within the intracellular volume with submicrometer accuracy. Results from investigations on cell swelling provoked by osmotic stimulation demonstrate the homogeneous stretching of the cytoskeleton network, and thus that the proposed method provides a new way for the quantitative 3D analysis of the dynamic intracellular morphology.

Ancillary