Rhodamine Spiroamides for Multicolor Single-Molecule Switching Fluorescent Nanoscopy
Rhodamine Spiroamides for Multicolor Single-Molecule Switching Fluorescent Nanoscopy
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DOI：
10.1002/chem.200901333
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发表时间：
2009-01-01
影响因子：
4.3
通讯作者：
Hell, Stefan W.
中科院分区：
文献类型：
--
作者：
Belov, Vladimir N.;Bossi, Mariano L.;Hell, Stefan W.
介绍了新型罗丹明螺酰胺的设计、合成和评价。这些分子在基于荧光单分子的随机开关的光学纳米显微镜中有应用。新标记可用于各种物体的（共）定位研究，它们的（相互）位置和形状可以以几十纳米的精度确定。由于罗丹明螺酰胺上存在各种官能团，因此实现了多色染色，良好的光活化，大量发射光子以及与氨基或巯基的选择性化学结合。刚性磺化的杂蒽片段与六元环、N，N'-二（2,2,2-三氟乙基）基团融合，以及与简单脂肪族螺酰胺残基结合的附加双键和磺酸基团提供了多色特性，并提高了罗丹明螺酰胺在光活化和生物偶联反应中的性能。将可光开关组件的两个基本部分——开关基团和荧光（报告）基团——结合在一个化学实体中，使这种方法具有进一步发展的吸引力。介绍了一系列罗丹明螺酰胺及其在单分子开关和定位显微镜中作为荧光探针应用的最相关性质的表征。具有纳米尺度分辨率的光学图像说明了标记在生物物体共定位和双光子激活技术与光学切片中的潜力。
The design, synthesis, and evaluation of new rhodamine spiroamides are described. These molecules have applications in optical nanoscopy based on random switching of the fluorescent single molecules. The new markers may be used in (co)localization studies of various objects and their (mutual) positions and shape can be determined with a precision of a few tens of nanometers. Multicolor staining, good photoactivation, a large number of emitted photons, and selective chemical binding with amino or thiol groups were achieved due to the presence of various functional groups on the rhodamine spiroamides. Rigidized sulfonated xanthene fragment fused with six-membered rings, N,N'-bis(2,2,2-trifluoroethyl) groups, and a combination of additional double bonds and sulfonic acid groups with simple aliphatic spiroamide residue provide multicolor properties and improve performance of the rhodamine spiroamides in photoactivation and bioconjugation reactions. Having both essential parts of the photoswitchable assembly-the switching and the fluorescent (reporter) groups-combined in one chemical entity make this approach attractive for further development. A series of rhodamine spiroamides is presented along with characterizations of their most relevant properties for application as fluorescent probes in single-molecule switching and localization microscopy. Optical images with resolutions on the nanometer scale illustrate the potential of the labels in the colocalization of biological objects and the two-photon activation technique with optical sectioning.