Fluorescent d-Amino Acids for Super-resolution Microscopy of the Bacterial Cell Wall

  • ACS Chem Biol. 2022 Sep 16;17(9):2418-2424. doi: 10.1021/acschembio.2c00496.
Chen Zhang  1 ,  Luc Reymond  2 ,  Ophélie Rutschmann  3 ,  Mischa A Meyer  1  3 ,  Julien Denereaz  4 ,  Jiangtao Qiao  5 ,  Faustine Ryckebusch  3 ,  Juliette Griffié  1 ,  Willi L Stepp  1 ,  Suliana Manley  1
Affiliations
  • 1. Institute of Physics, School of Basic Sciences, Swiss Federal Institute of Technology Lausanne (EPFL), Lausanne 1015, Switzerland.
  • 2. Biomolecular Screening Core Facility, Swiss Federal Institute of Technology Lausanne (EPFL), Lausanne 1015, Switzerland.
  • 3. Global Health Institute, School of Life Sciences, Swiss Federal Institute of Technology Lausanne (EPFL), Lausanne 1015, Switzerland.
  • 4. Department of Fundamental Microbiology, Faculty of Biology and Medicine, University of Lausanne (UNIL), Lausanne 1015, Switzerland.
  • 5. Environmental Engineering Institute, School of Architecture, Civil and Environmental Engineering, Swiss Federal Institute of Technology Lausanne (EPFL), Lausanne 1015, Switzerland.
PMID: 35994360 DOI: 10.1021/acschembio.2c00496
Abstract

Fluorescent d-amino acids (FDAAs) have previously been developed to enable in situ highlighting of locations of Bacterial cell wall growth. Most Bacterial cells lie at the edge of the diffraction limit of visible light; thus, resolving the precise details of peptidoglycan (PG) biosynthesis requires super-resolution microscopy after probe incorporation. Single molecule localization microscopy (SMLM) has stringent requirements on the fluorophore photophysical properties and therefore has remained challenging in this context. Here, we report the synthesis and characterization of new FDAAs compatible with one-step labeling and SMLM imaging. We demonstrate the incorporation of our probes and their utility for visualizing PG at the nanoscale in Gram-negative, Gram-positive, and mycobacteria species. This improved FDAA toolkit will endow researchers with a nanoscale perspective on the spatial distribution of PG biosynthesis for a broad range of Bacterial species.

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