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Abstract：
The use of photoactivatable dyes in STED microscopy has so far been limited by two-photon activation through the STED beam and by the fact that photoactivatable dyes are poorly solvable in water. Herein, we report ONB-2SiR, a fluorophore that can be both photoactivated in the UV and specifically de-excited by STED at 775 nm. Likewise, we introduce a conjugation and purification protocol to effectively label primary and secondary antibodies with moderately water-soluble dyes. Greatly reducing dye aggregation, our technique provides a defined and tunable degree of labeling, and improves the imaging performance of dye conjugates in general.
摘要：
迄今为止,在STED显微镜中使用光活化染料受到通过STED光束的双光子活化以及光活化染料在水中难溶解的事实的限制。在这里，我们报道了ONB-2SiR,一种荧光团，既可以在UV中光活化，又可以通过STED在775nm处特异性去激发。同样，我们引入了缀合和纯化方案,用中等水溶性染料有效标记一级和二级抗体.大大减少染料聚集，我们的技术提供了一个定义和可调的标记程度，总体上提高了染料偶联物的成像性能。
