The Power of Two: Covalent Coupling of Photostabilizers for Fluorescence Applications
详细信息 查看全文
- 作者：Jasper H. M. van der Velde ; Jens Oelerich ; Jingyi Huang ; Jochem H. Smit ; Matthias Hiermaier ; Evelyn Ploetz ; Andreas Herrmann ; Gerard Roelfes ; Thorben Cordes
- 刊名：The Journal of Physical Chemistry Letters
- 出版年：2014
- 出版时间：November 6, 2014
- 年：2014
- 卷：5
- 期：21
- 页码：3792-3798
- 全文大小：473K
- ISSN：1948-7185
文摘
Fluorescence is a versatile tool for spectroscopic investigations and imaging of dynamic processes and structures across various scientific disciplines. The photophysical performance, that is, signal stability and signal duration, of the employed fluorophores is a major limiting factor. In this Letter, we propose a general concept to covalently link molecules, which are known for their positive effect in photostabilization, to form a combined photostabilizer with new properties. The direct linkage of two (or more) photostabilizers will allow one to obtain combined or synergetic effects in fluorophore stabilization and can simplify the preparation of imaging buffers that would otherwise require a mixture of photostabilizers for optimal performance. This concept was explored by synthesizing a molecule with a reducing and oxidizing moiety that is referred to as internal ROXS or 鈥渋ROXS鈥? Using single-molecule fluorescence microscopy, inter- and intramolecular healing of iROXS was observed, that is, strongly reduced blinking and increased photostability of the cyanine fluorophore Cy5. Moreover, it is shown that a covalently coupled photostabilizer can replace a mixture of molecules needed to make a functional photostabilizing ROXS buffer and might hence represent the new standard for defined and reproducible imaging conditions in single-molecule experiments. In self-healing fluorophores with intramolecular triplet-state quenching, an unprecedented photostability increase of >100-fold was obtained when using iROXS, which is even competitive with solution-based healing. Control experiments show that the oxidizing part of the iROXS molecule, an aromatic nitro group, dominates the healing process. The suggested synthetic concept and the proof-of-concept experiments represent the starting point for the quest to identify optimal combinations of linked photostabilizers for various fluorescence applications.