Kojima H, Hirotani M, Nakatsubo N, Kikuchi K, Urano Y, Higuchi T, Hirata Y, Nagano T
Graduate School of Pharmaceutical Sciences, and Second Department of Internal Medicine, Faculty of Medicine, University of Tokyo, Japan.
Anal Chem. 2001 May 1;73(9):1967-73. doi: 10.1021/ac001136i.
Diaminofluoresceins are widely used for detection and imaging of nitric oxide (NO), but for biological applications, they have the disadvantages that the fluorescence of the fluorescein chromophore is pH-sensitive and overlaps the autofluorescence of cells. We have developed a membrane-permeable fluorescent indicator for NO based on the rhodamine chromophore, DAR-4M AM, which can be excited with 550-nm light. The fluorescence quantum yield of the product after reaction with NO is 840 times higher than that of DAR-4M. The detection limit of NO was 7 nM, and the fluorescence showed no pH dependency above pH 4. DAR-4M AM was successfully applied to practical bioimaging of NO produced in bovine aortic endothelial cells.
二氨基荧光素被广泛用于一氧化氮（NO）的检测和成像，但对于生物应用而言，它们存在荧光素发色团的荧光对pH敏感且与细胞自发荧光重叠的缺点。我们基于罗丹明发色团开发了一种用于NO的膜渗透性荧光指示剂DAR-4M AM，它可以用550纳米的光激发。与NO反应后产物的荧光量子产率比DAR-4M高840倍。NO的检测限为7纳摩尔，并且在pH值高于4时荧光不显示pH依赖性。DAR-4M AM已成功应用于牛主动脉内皮细胞中产生的NO的实际生物成像。
