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Quantitative multicolor subdiffraction imaging of bacterial protein ultrastructures in three dimensions.

ABSTRACT: 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.

SUBMITTER: Gahlmann A

PROVIDER: S-EPMC3599789 | biostudies-literature | 2013 Mar

REPOSITORIES: biostudies-literature

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Quantitative multicolor subdiffraction imaging of bacterial protein ultrastructures in three dimensions.

Gahlmann Andreas A Ptacin Jerod L JL Grover Ginni G Quirin Sean S von Diezmann Lexy L Lee Marissa K MK Backlund Mikael P MP Shapiro Lucy L Piestun Rafael R Moerner W E WE

Nano letters 20130225 3

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 differ ...[more]

PMID: 23414562

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