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Nano Lett. 2013 Mar 13;13(3):987-93. doi: 10.1021/nl304071h. Epub 2013 Feb 25.

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

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1
Department of Chemistry, Stanford University, Stanford, California 94305, United States.

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.

PMID:
23414562
PMCID:
PMC3599789
DOI:
10.1021/nl304071h
[Indexed for MEDLINE]
Free PMC Article
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