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Volume 166, Issue 12

An OregonGreen488-labelled -amino acid for visualizing peptidoglycan by super-resolution STED nanoscopy Open Access

Abstract

Fluorescent -amino acids (FDAAs) are molecular probes that are widely used for labelling the peptidoglycan layer of bacteria. When added to growing cells they are incorporated into the stem peptide by a transpeptidase reaction, allowing the timing and localization of peptidoglycan synthesis to be determined by fluorescence microscopy. Herein we describe the chemical synthesis of an OregonGreen488-labelled FDAA (OGDA). We also demonstrate that OGDA can be efficiently incorporated into the PG of Gram-positive and some Gram-negative bacteria, and imaged by super-resolution stimulated emission depletion (STED) nanoscopy at a resolution well below 100 nm.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): FDAA , FLDA , OGDA , OregonGreen488 , peptidoglycan and transpeptidase
Funding
This study was supported by the:
  • Knut och Alice Wallenbergs Stiftelse
    • Principle Award Recipient: GöranWidmalm
  • Vetenskapsrådet (Award 2017-03703)
    • Principle Award Recipient: GöranWidmalm
  • Japan Society for the Promotion of Science (Award JP17K15694)
    • Principle Award Recipient: BillSöderström
  • Vetenskapsrådet
    • Principle Award Recipient: DanielDaley
  • Carl Tryggers Stiftelse för Vetenskaplig Forskning (SE)
    • Principle Award Recipient: DanielDaley
© 2020 The Authors
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2020-11-25
2024-04-16
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An OregonGreen488-labelled d-amino acid for visualizing peptidoglycan by super-resolution STED nanoscopy
Microbiology 166, 1129 (2020); https://doi.org/10.1099/mic.0.000996
/content/journal/micro/10.1099/mic.0.000996
/content/journal/micro/10.1099/mic.0.000996
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