Novel application of low pH-dependent fluorescent dyes to examine colitis

BMC Gastroenterology

Technical Advance

Takafumi Ando¹  Osamu Watanabe¹  Hidemi Goto¹  Kazuhiro Ishiguro² 

[1] Department of Gastroenterology, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, 466-8550, Aichi, Japan;Molecular Biology and Pathogenesis of Gastroenterology, Nagoya University School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, 466-8550, Aichi, Japan;

关键词: Methylene Blue;    Goblet Cell;    Topical Application;    Colonic Mucosa;    Dextran Sulfate Sodium;   

DOI  :  10.1186/1471-230X-10-4

 received in 2009-08-27, accepted in 2010-01-15,  发布年份 2010

来源: Springer

【 摘 要 】

BackgroundEndoscopy capable of fluorescence observation provides histological information on gastrointestinal lesions. We explored the novel application of low pH-dependent fluorescent dyes for fluorescence observation of crypt structure and inflammatory cell infiltration in the colon.MethodsLow pH-dependent fluorescent dyes were applied to the colonic mucosa of normal mice for observation under fluorescence stereomicroscopy system. We also examined mouse models of colitis, which were induced by trinitrobenzenesulfonic acid, dextran sulfate sodium or interleukin-10 deficiency.ResultsTopical application of low pH-dependent fluorescent dyes revealed crypts as ring-shaped fluorescent stains by visualizing the mucin granules of goblet cells. Because of the minimal fluorescence intensity of the low pH-dependent fluorescent dyes in phosphate-buffered saline, it was not necessary to wash the mucosa before the fluorescence observation. 4-Nitro-7-piperazino-2,1,3-benzoxadiazole (NBD-PZ) was quicker to achieve complete staining (three minutes) than LysoSensor Green DND-153 and DND-189 (20 minutes). In each type of colitis, NBD-PZ revealed the destruction of the crypts as the disappearance of the ring-shaped fluorescent stains and the infiltration of inflammatory cells as the aggregation of punctate fluorescent stains through visualization of lysosomes.ConclusionsLow pH-dependent fluorescent dyes, especially NBD-PZ, are suitable for topical application to the colonic mucosa and have characteristics that allow for the histological examination of colitis.

【 授权许可】

CC BY   
© Ishiguro et al; licensee BioMed Central Ltd. 2010