Phosphine Quenching of Cyanine Dyes as a Versatile Tool for Fluorescence Microscopy
2013
Vaughan, Joshua C. | Dempsey, Graham T. | Sun, Eileen | Zhuang, Xiaowei
We report that the cyanine dye Cy5 and several of its structural relatives are reversibly quenched by the phosphine tris(2-carboxyethyl)phosphine (TCEP). Using Cy5 as a model, we show that the quenching reaction occurs by 1,4-addition of the phosphine to the polymethine bridge of Cy5 to form a covalent adduct. Illumination with UV light dissociates the adduct and returns the dye to the fluorescent state. We demonstrate that TCEP quenching can be used for super-resolution imaging as well as for other applications, such as differentiating between molecules inside and outside the cell.
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