Photoinduced formation of reversible dye radicals and their impact on super-resolution imaging
Photoinduced formation of reversible dye radicals and their impact on super-resolution imaging
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DOI：
10.1039/c0pp00317d
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发表时间：
2011-01-01
影响因子：
3.1
通讯作者：
Sauer, Markus
中科院分区：
文献类型：
--
作者：
van de Linde, Sebastian;Krstic, Ivan;Sauer, Markus
有机染料的自由基离子是高活性物种，并且已经通过瞬态吸收光谱和脉冲辐解实验在贫氧溶液中研究了几十年。在这里，我们通过连续波EPR，吸收和荧光实验表明，三重态的罗丹明染料可以被巯基光还原，形成稳定的自由基阴离子在水溶液中的寿命长达几个小时。我们的数据表明，减少三重态和光诱导氧化的活性中间体的氧代表了可逆的光开关的染料在含巯基的水溶液中的一般机制，突出了分子氧的超分辨率荧光成像的关键作用。由于细胞含有巯基谷胱甘肽在毫摩尔浓度和活性氧物种形成的副产品，我们的研究结果是活细胞荧光显微镜的后果。
Radical ions of organic dyes are highly reactive species and have been studied for decades by transient absorption spectroscopy and pulse radiolysis experiments in oxygen-depleted solution. Here we show by continuous wave EPR, absorption, and fluorescence experiments that the triplet state of rhodamine dyes can be photoreduced by thiols to form stable radical anions in aqueous solution with a lifetime of up to several hours. Our data demonstrate that reduction of the triplet state and photoinduced oxidation of reactive intermediates by oxygen represents a general mechanism for reversible photoswitching of dyes in aqueous thiol-containing solutions highlighting the key role of molecular oxygen for super-resolution fluorescence imaging. Since cells contain the thiol glutathione at millimolar concentrations and reactive oxygen species are formed as side products our findings are of consequence for live cell fluorescence microscopy.