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

• Published in: BioTechniques Vol. 45; no. 4; pp. 465 - 468
• Main Authors: Ishiguro, Kazuhiro, Ando, Takafumi, Goto, Hidemi
• Format: Journal Article
• Language: English
• Published: Natick, MA Future Science Ltd 01.10.2008; Eaton; Taylor & Francis Group
• Subjects: Animals; Biological and medical sciences; Cell Line; Cell Survival; Cytological Techniques; Diverse techniques; Fundamental and applied biological sciences. Psychology; Humans; Lysosomes - metabolism; Mice; Microscopy, Fluorescence; Molecular and cellular biology; Oxadiazoles - chemistry; Oxadiazoles - economics; Oxadiazoles - metabolism; Piperazines - chemistry; Piperazines - economics; Piperazines - metabolism; Rats; Application; Lysosome
• ISSN: 0736-6205; 1940-9818
• 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-CH CH NH 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 ≤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.