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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Time-resolved And Two-photon Emission Imaging Microscopy Of Live Cells With Inert Platinum Complexes
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Time-resolved And Two-photon Emission Imaging Microscopy Of Live Cells With Inert Platinum Complexes

机译：具有惰性铂络合物的活细胞的时间分辨和双光子发射成像显微镜

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This work explores time-resolved emission imaging microscopy (TREM) for noninvasive imaging and mapping of live cells on a hitherto uncharted microsecond time scale. Simple robust molecules for this purpose have long been sought. We have developed highly emissive, synthetically versatile, and photostable platinum(II) complexes that make TREM a practicable reality. [PtLCl], {HL = 1,3-di(2-pyridyl)benzene and derivatives}, are charge-neutral, small molecules that have low cytotoxicity and accumulate intracellularly within a remarkably short incubation time of 5 min, apparently under diffusion control. Their microsecond lifetimes and emission quantum yields of up to 70% are exceptionally high for transition metal complexes and permit the application of TREM to be demonstrated in a range of live cell types-normal human dermal fibroblast, neoplas-tic C8161 and CHO cells. [PtLCl] are thus likely to be suitable emission labels for any eukaryotic cell types. The high photostability of [PtLCl] under intense prolonged irradiation has allowed the development of tissue-friendly NIR two-photon excitation (TPE) in conjunction with transition metal complexes in live cells. A combination of confocal one-photon excitation, nonlinear TPE, and microsecond time-resolved imaging has revealed (ⅰ) preferential localization of the complexes to intracellular nucleic acid structures, in particular the nucleoli and (ⅱ) the possibility of measuring intracellular emission lifetimes in the microsecond range. The combination of TREM, TPE, and Pt(II) complexes will be a powerful tool for investigating intracellular processes in vivo, because the long lifetimes allow discrimination from autofluo-rescence and open up the use of commonplace technology.
机译：这项工作探索了时间分辨发射成像显微镜（TREM），以迄今未知的微秒时间尺度对活细胞进行无创成像和标测。长期以来一直寻求用于该目的的简单的坚固分子。我们已经开发出具有高发射性，合成用途广泛且耐光的铂（II）配合物，使TREM成为现实。 [PtLCl]，{HL = 1,3-二（2-吡啶基）苯及其衍生物}，是电荷中性的小分子，具有低细胞毒性，并在非常短的5分钟孵育时间内在细胞内蓄积，显然处于扩散控制之下。它们的微秒寿命和高达70％的发射量子产率对于过渡金属络合物而言异常高，并允许TREM在一系列活细胞类型中正常人的真皮成纤维细胞，赘生性C8161和CHO细胞中得到证明。因此，[PtLCl]可能是任何真核细胞类型的合适发射标记。 [PtLCl]在高强度长时间照射下的高光稳定性已允许在活细胞中与过渡金属配合物一起发展组织友好型近红外双光子激发（TPE）。共聚焦单光子激发，非线性TPE和微秒时间分辨成像的组合显示（ⅰ）络合物优先定位于细胞内核酸结构（特别是核仁），以及（ⅱ）测量细胞中细胞内发射寿命的可能性。微秒范围。 TREM，TPE和Pt（II）配合物的组合将成为研究体内细胞内过程的有力工具，因为其长寿命可以区分自发荧光并开辟了普通技术的使用。

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