Real-time monitoring of endogenous cysteine levels in living cells using a CD-based ratiometric fluorescent nanoprobe

Anal Bioanal Chem. 2018 Jul;410(18):4379-4386. doi: 10.1007/s00216-018-1091-x. Epub 2018 Apr 30.

Abstract

A simple and readily available fluorescent probe is needed for the real-time monitoring of endogenous cysteine (Cys) levels in living cells, as such a probe could be used to study the role of Cys in related diseases. Herein, we report the first fluorescent probe based on carbon dots (CDs-FITA) for the selective and ratiometric imaging of endogenous Cys in live cells. In this ratiometric fluorescent probe, a fluorescein derivative (FITA) that recognizes Cys is covalently linked to the surfaces of carbon dots (CDs); employing CDs greatly improves the water solubility of the probe. Acrylate on FITA is selectively cleaved by Cys in aqueous solution under mild conditions, leading to a dramatic increase in the fluorescence from fluorescein. The probe therefore allows the highly selective ratiometric fluorescent detection of Cys even in the presence of various interferents. The as-prepared CDs-FITA showed excellent performance when applied to detect Cys in blood serum. In addition, due to its negligible cytotoxicity, the CDs-FITA can also be utilized for the real-time monitoring of endogenous cysteine (Cys) levels in living cells. Graphical abstract Illustration of the CD-based probe for Cys imaging in living cells.

Keywords: Carbon dots; Cys; Fluorescein; Living cell imaging; Ratiometric fluorescence.

MeSH terms

  • Carbon / chemistry*
  • Carbon-13 Magnetic Resonance Spectroscopy
  • Cysteine / blood
  • Cysteine / metabolism*
  • Fluorescein / chemistry
  • Fluorescent Dyes / chemistry*
  • HeLa Cells
  • Humans
  • Nanoparticles / chemistry*
  • Proton Magnetic Resonance Spectroscopy
  • Solubility
  • Spectrometry, Fluorescence / methods*
  • Spectrometry, Mass, Electrospray Ionization
  • Water / chemistry

Substances

  • Fluorescent Dyes
  • Water
  • Carbon
  • Cysteine
  • Fluorescein