Quantitative multicolor subdiffraction imaging of bacterial protein ultrastructures in three dimensions

Andreas Gahlmann, Jerod L. Ptacin, Ginni Grover, Sean Quirin, Alexander R.S. Von Diezmann, Marissa K. Lee, Mikael P. Backlund, Lucy Shapiro, Rafael Piestun, W. E. Moerner

Research output: Contribution to journalArticlepeer-review


We demonstrate quantitative multicolor three-dimensional (3D) subdiffraction imaging of the structural arrangement of fluorescent protein fusions in living Caulobacter crescentus bacteria. Given single-molecule localization precisions of 20-40 nm, a flexible locally weighted image registration algorithm is critical to accurately combine the super-resolution data with <10 nm error. Surface-relief dielectric phase masks implement a double-helix response at two wavelengths to distinguish two different fluorescent labels and to quantitatively and precisely localize them relative to each other in 3D.

Original languageEnglish (US)
Pages (from-to)987-993
Number of pages7
JournalNano letters
Issue number3
StatePublished - Mar 13 2013
Externally publishedYes


  • 3D imaging
  • biophysics
  • Fluorescence
  • image registration
  • live-cell imaging
  • super-resolution

ASJC Scopus subject areas

  • Bioengineering
  • Chemistry(all)
  • Materials Science(all)
  • Condensed Matter Physics
  • Mechanical Engineering


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