Near-infrared luminescent metallacrowns for combined in vitro cell fixation and counter staining.
Near-infrared luminescent metallacrowns for combined in vitro cell fixation and counter staining.
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DOI：
10.1039/c7sc01872j
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发表时间：
2017-09-01
期刊：
影响因子：
8.4
通讯作者：
Petoud S
中科院分区：
文献类型：
--
作者：
Martinić I;Eliseeva SV;Nguyen TN;Foucher F;Gosset D;Westall F;Pecoraro VL;Petoud S
在UV-A光照射下，使用含吡嗪异羟肟酸的LnIII/ZnII MCs实现细胞固定和近红外对抗染色。细胞固定是保存细胞形态的基本方法，允许用荧光探针靶向和标记生物分子。更有效的荧光标记的关键要求之一是保持细胞形态，这通常需要几种固定技术的组合。此外，反染色的使用对于提高荧光探针的对比度通常是必不可少的。目前的试剂有很大的局限性，如对光漂白的抵抗力低，对微环境的变化敏感。发光的Ln3+‘封装夹层’金属冠(MC)克服了这些缺点，提供了互补的优势。特别是，它们发射出尖锐的发射波段，在激发和发射波长之间具有很大的差异，并且不会光漂白。在本研究中，使用了吡嗪异羟肟酸形成的MC(Ln3+[Zn(II)MCpyzHA]，Ln3+=Yb，ND)，结合近红外(NIR)对数染色和固定剂对HeLa细胞进行UV-A光照射，最初5分钟。用拉曼光谱评价细胞固定的有效性和质量。在不同的实验条件下，包括在染色细胞的悬浮液中，分析了这些MC的近红外发光特性。此外，Ln3+[Zn(II)MCpyzHA]在近红外区域的高发射强度使这些MC可以用于安装在常规荧光显微镜上的标准CCD相机的成像。最后，发射近红外的Ln3+[Zn(II)MCpyzHA]化合物在单个分子内结合了细胞固定和染色能力、良好的光稳定性和发射光谱对局部微环境的最小敏感度等特点，在建立具有单一生物分布的下一代显像剂方面具有很大的潜力。
Combined cell fixation and near-infrared counter staining was achieved using LnIII/ZnII MCs with pyrazinehydroxamic acid upon illumination with UV-A light. Cell fixation is an essential approach for preserving cell morphology, allowing the targeting and labelling of biomolecules with fluorescent probes. One of the key requirements for more efficient fluorescent labelling is the preservation of cell morphology, which usually requires a combination of several fixation techniques. In addition, the use of a counter stain is often essential to improve the contrast of the fluorescent probes. Current agents possess significant limitations, such as low resistance toward photobleaching and sensitivity to changes in the microenvironment. Luminescent Ln3+ ‘encapsulated sandwich’ metallacrowns (MCs) overcome these drawbacks and offer complementary advantages. In particular, they emit sharp emission bands, possess a large difference between excitation and emission wavelengths and do not photobleach. Herein, MCs formed with pyrazinehydroxamic acid (Ln3+[Zn(ii)MCpyzHA], Ln3+ = Yb, Nd) were used, combined with near-infrared (NIR) counter staining and fixation agents for HeLa cells upon an initial five minute exposure to UV-A light. The validity and quality of the cell fixation were assessed with Raman spectroscopy. Analysis of the NIR luminescence properties of these MCs was performed under different experimental conditions, including in a suspension of stained cells. Moreover, the high emission intensity of Ln3+[Zn(ii)MCpyzHA] in the NIR region allows these MCs to be used for imaging with standard CCD cameras installed on routine fluorescence microscopes. Finally, the NIR-emitting Ln3+[Zn(ii)MCpyzHA] compounds combine, within a single molecule, features such as cell fixation and staining abilities, good photostability and minimal sensitivity of the emission bands to the local microenvironment, and they are highly promising for establishing the next generation of imaging agents with a single biodistribution.
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