Sensitivity evaluation of NBD-SCN towards cysteine/homocysteine and its bioimaging applications

Biosens Bioelectron. 2014 Jun 15:56:117-23. doi: 10.1016/j.bios.2014.01.009. Epub 2014 Jan 11.

Abstract

A push-pull fluorogenic reagent, NBD-SCN, was applied for specific detection of cysteine (Cys) and homocysteine (Hcy). Replacing thiocyanato group with Cys/Hcy increased the push-pull characteristic of the probe and resulted in emission of fluorescence. The fluorescent response of the probe toward Cys/Hcy was significantly higher than toward glutathione and other amino acids. The probe showed a 470- and 745-fold fluorescence enhancement at 550 nm and detection limit of 2.99 and 1.43 nM for Cys and Hcy, respectively. Time-dependent fluorescence assays showed that the fluorescence intensity reached a plateau within 20s after addition of Cys and within 10 min after addition of Hcy. Furthermore, the fluorescence images of Cys/Hcy in Raw 264.7 cells were obtained after adding this probe to the cells. These results indicate that NBD-SCN not only possesses good selectivity and sensitivity for Cys/Hcy but also can penetrate cells for Cys/Hcy bioimaging.

Keywords: Cysteine; Fluorescent probe; Homocysteine; ICT; NBD.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biosensing Techniques / methods
  • Cell Line
  • Cysteine / analysis*
  • Fluorescent Dyes / chemistry*
  • Homocysteine / analysis*
  • Limit of Detection
  • Macrophages / chemistry
  • Macrophages / cytology
  • Mice
  • Microscopy, Confocal / methods
  • Optical Imaging / methods*
  • Spectrometry, Fluorescence / methods

Substances

  • Fluorescent Dyes
  • Homocysteine
  • Cysteine