Letter pubs.acs.org/NanoLett
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,‡ and W. E. Moerner*,† †
Department of Chemistry, Stanford University, Stanford, California 94305, United States Department of Electrical, Computer, and Energy Engineering, University of Colorado at Boulder, 425 UCB, Boulder, Colorado 80309, United States § Department of Developmental Biology, Stanford University School of Medicine, Beckman Center, Stanford, California 94305, United States ‡
S Supporting Information *
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
