Novel application of 4-nitro-7-(1-piperazinyl)-2,1,3-benzoxadiazole to visualize lysosomes in live cells

Biotechniques. 2008 Oct;45(4):465, 467-8. doi: 10.2144/000112912.


4-Nitro-7-(1-piperazinyl)-2,1,3-benzoxadiazole (NBD-PZ) reacts with carboxylic acids in the presence of condensing agents and is utilized for the fluorescence detection of the generated derivatives in high performance liquid chromatography or capillary electro-phoresis. Although the fluorescence intensity of derivatives of NBD-PZ-CH2CH2NH2 with carboxylic acids is elevated at low pH, the pH-dependent fluorescence of NBD-PZ itself has not yet been investigated. In this study, we determined the fluorescence spectra of NBD-PZ at various pH and found that the fluorescence intensity of NBD-PZ was elevated dramatically at pH <or=7.0. When NBD-PZ was applied to live HGC-27 cells, CW-2 cells, IEC-18 cells, and RAW264.7 cells, we observed that NBD-PZ clearly visualized lysosomes under fluorescence microscopy within 3 min. These findings indicate a novel application of NBD-PZ to visualize lysosomes in live cells, a strategy that offers substantial cost savings over commercially available lysosomal probes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Cell Survival
  • Cytological Techniques*
  • Humans
  • Lysosomes / metabolism*
  • Mice
  • Microscopy, Fluorescence
  • Oxadiazoles / chemistry
  • Oxadiazoles / economics
  • Oxadiazoles / metabolism*
  • Piperazines / chemistry
  • Piperazines / economics
  • Piperazines / metabolism*
  • Rats


  • 4-nitro-7-piperazino-2,1,3-benzoxadiazole
  • Oxadiazoles
  • Piperazines