Photoactivatable Fluorophore for Stimulated Emission Depletion (STED) Microscopy and Bioconjugation Technique for Hydrophobic Labels,Chemistry - A European Journal

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Photoactivatable Fluorophore for Stimulated Emission Depletion (STED) Microscopy and Bioconjugation Technique for Hydrophobic Labels
Chemistry - A European Journal ( IF 3.9 ) Pub Date : 2020-10-23 , DOI: 10.1002/chem.202004645
Michael Weber ₁ , Taukeer A Khan ₁ , Lukas J Patalag _{1,

2} , Mariano Bossi ₃ , Marcel Leutenegger ₁ , Vladimir N Belov ₁ , Stefan W Hell _{1,

3}

Affiliation

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.

中文翻译：

用于受激发射损耗 (STED) 显微镜和疏水标记生物共轭技术的光活化荧光团

迄今为止，光活化染料在 STED 显微镜中的使用受到通过 STED 光束的双光子激活以及光活化染料在水中溶解性较差这一事实的限制。在此，我们报道了 ONB-2SiR，一种荧光团，既可以在 UV 中光激活，又可以在 775 nm 处通过 STED 特异性去激发。同样，我们引入了缀合和纯化方案，以使用中等水溶性染料有效地标记一抗和二抗。我们的技术大大减少了染料聚集，提供了明确且可调的标记程度，并总体上提高了染料缀合物的成像性能。

更新日期：2020-10-23

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