Novel application of 4-nitro-7-(1-piperazinyl)-2,1,3-benzoxadiazole to visualize lysosomes in live cells.
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.
Molecular Biology and Pathogenesis of Gastroenterology, Nagoya University School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, Aichi 466-8550, Japan. kio@med.nagoya-u.ac.jp
Kazuhiro
K
Ishiguro
Takafumi
T
Ando
Hidemi
H
Goto
D000818:Animals; D002460:Cell Line; D002470:Cell Survival; D003584:Cytological Techniques; D006801:Humans; D008247:Lysosomes / Q000378:metabolism*; D051379:Mice; D008856:Microscopy, Fluorescence; D010069:Oxadiazoles / Q000737:chemistry / Q000191:economics / Q000378:metabolism*; D010879:Piperazines / Q000737:chemistry / Q000191:economics / Q000378:metabolism*; D051381:Rats
C432126:4-nitro-7-piperazino-2,1,3-benzoxadiazole; D010069:Oxadiazoles; D010879:Piperazines