3D single molecule super-resolution imaging of cellular samples using MUM

POSTER

Abstract

Localization based super-resolution imaging techniques (e.g., PALM, fPALM, STORM, etc.) represent a powerful tool to image single molecules at nanoscale resolution in two dimensions. However, the extension of these techniques to three dimensions poses several technical challenges, foremost being the poor depth discrimination inherent to conventional microscopes which is typically used by these techniques. Previously, we developed an imaging modality, multifocal plane microscopy (MUM), which overcomes the poor depth discrimination capability of conventional microscopes. We also introduced a 3D localization algorithm MUMLA and demonstrated experimentally that it provides the best possible accuracy with which the 3D position of single molecules can be determined. Here, we extend the application of MUM and MUMLA for super-resolution imaging and demonstrate 3D imaging of single molecules at nanometer scale resolution in a cellular sample.

Authors

  • Andrea Grosso

    • Department of Electrical Engineering, UT Dallas
  • Anish Abraham

    • Dept. of Immunology, UT Southwestern
    • Department of Electrical Engineering, UT Dallas
  • Jeffrey Kang

    • Dept. of Immunology, UT Southwestern Medical Center, Dallas, TX
  • Jerry Chao

    • Dept. of Immunology, UT Southwestern Medical Center, Dallas, TX
  • Ramraj Velmurugan

    • Dept. of Immunology, UT Southwestern Medical Center, Dallas, TX
  • Sripad Ram

    • Dept. of Immunology, UT Southwestern Medical Center, Dallas, TX
  • Stephen Anthony

    • Dept. of Immunology, UT Southwestern Medical Center, Dallas, TX
  • Zhuo Gan

    • Dept. of Immunology, UT Southwestern Medical Center, Dallas, TX
  • E. Sally Ward

    • Dept. of Immunology, UT Southwestern
    • Department of Immunology, University of Texas Southwestern Medical Center, Dallas, Texas
    • Dept. of Immunology, UT Southwestern Medical Center, Dallas, TX
    • University of Texas Southwestern Medical Center
  • Raimund J. Ober

    • Department of Electrical Engineering, University of Texas at Dallas, Richardson, Texas
    • Department of Electrical Engineering, UT Dallas
    • University of Texas at Dallas