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Fluorescent Copolymers for Bacterial Bioimaging and Viability Detection.

Yang Si1,2, Chloé Grazon1, Gilles Clavier1

  • 1Université Paris-Saclay, ENS Paris-Saclay, CNRS, PPSM, 91190 Gif-sur-Yvette, France.

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|August 14, 2020
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Summary

We developed a novel, nontoxic green fluorescent polymer chain (GFPC) for multicolor bioimaging and real-time bacterial viability detection. This biocompatible fluorescent label is stable in complex media and enables accurate live/dead bacterial quantification.

Keywords:
RAFTbacteriabioimagingflow cytometryfluorescencepolymerviability detection

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Area of Science:

  • Microbiology
  • Polymer Chemistry
  • Biotechnology

Background:

  • High photostability and biocompatibility are crucial for microbiological imaging and detection.
  • Existing fluorescent labels often lack these properties, limiting their application.

Purpose of the Study:

  • To develop a novel, nontoxic, and water-soluble green fluorescent polymer chain (GFPC) for multicolor bioimaging.
  • To assess GFPC's utility for real-time bacterial viability determination.

Main Methods:

  • Synthesis of GFPC via one-pot reversible addition-fragmentation chain-transfer (RAFT) polymerization.
  • Evaluation of GFPC's biocompatibility, stability in complex media, and bacterial growth influence.
  • Application of GFPC for bacterial cytoplasm labeling and multicolor bioimaging.
  • Development of a flow cytometry protocol using GFPC and propidium iodide (PI) for live/dead bacterial discrimination.

Main Results:

  • GFPC was successfully synthesized and demonstrated to be nontoxic.
  • GFPC exhibited high photostability and remained stable in complex growth media with bacteria for several hours.
  • GFPC effectively labeled bacterial cytoplasm and did not impede bacterial growth.
  • A rapid and reliable flow cytometry protocol was established to quantify live and dead bacteria in real time using GFPC and PI.

Conclusions:

  • The developed green fluorescent polymer chain (GFPC) is a promising tool for microbiological applications.
  • GFPC offers high biocompatibility and photostability, enabling advanced multicolor bioimaging.
  • GFPC facilitates real-time differentiation and quantification of live and dead bacteria, advancing diagnostic capabilities.