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Fluorescent Polymer-AS1411-Aptamer Probe for dSTORM Super-Resolution Imaging of Endogenous Nucleolin.

Laura Fabre1, Corentin Rousset2, Karine Monier2

  • 1Univ Lyon, CNRS, UMR 5223, Ingénierie des Matériaux Polymères, Université Claude Bernard Lyon 1, INSA Lyon, F-69622 Villeurbanne Cédex, France.

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|May 13, 2022
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Summary

Researchers developed small, bright polymer-aptamer probes for super-resolution microscopy. These probes enable precise detection of nucleolin (a key protein) in cells, offering an alternative to traditional methods.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Nanotechnology

Background:

  • Nucleolin is a vital protein involved in numerous cellular functions.
  • Accurate localization of nucleolin is crucial for understanding its roles.
  • Super-resolution microscopy techniques offer enhanced cellular visualization.

Purpose of the Study:

  • To develop novel polymer-aptamer probes for precise nucleolin localization.
  • To adapt these probes for stochastic optical reconstruction microscopy (dSTORM).
  • To evaluate the probes' performance in vitro and in cellulo.

Main Methods:

  • Synthesis of fluorescent polymer chains via RAFT polymerization.
  • Conjugation of polymers with AS1411 DNA aptamer using click chemistry (SPAAC).
  • Evaluation of probe specificity, brightness, and size for dSTORM imaging.

Main Results:

  • Developed small, bright fluorescent polymer-aptamer probes.
  • Confirmed retention of nucleolin recognition and specificity.
  • Demonstrated selective and super-resolved (10-20 nm) detection of nucleolin via dSTORM.

Conclusions:

  • Polymer-aptamer probes are effective for super-resolution imaging of nucleolin.
  • These probes offer a promising alternative to immunofluorescence for nanoscopy.
  • The developed probes enable precise visualization of nucleolin in cellular environments.