Using AIE Luminogen for Long-term and Low-background Three-Photon Microscopic Functional Bioimaging.
Using AIE Luminogen for Long-term and Low-background Three-Photon Microscopic Functional Bioimaging.
复制标题
使用AIE Luminogen进行长期和低背景的三光子显微镜功能生物成像。
DOI：
10.1038/srep15189
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发表时间：
2015-10-15
影响因子：
4.6
通讯作者：
He S
中科院分区：
文献类型：
--
作者：
Zhu Z;Leung CW;Zhao X;Wang Y;Qian J;Tang BZ;He S
荧光探针是最常用的生物成像标记物之一，用于监测活细胞的代谢过程。然而，长时间的光激发往往会导致荧光探针的光漂白、不可避免的自发荧光以及细胞的光损伤。为了克服这些限制，我们合成了一种具有聚集诱导发射（AIE）特性的光稳定发光体TPE-TPP，并在1020 nm飞秒激光激发下实现了其三光子成像。以TPE-TPP为荧光探针，在1020 nm激发下的三光子显微镜观察显示，TPE-TPP对HeLa细胞的光损伤很小，自发荧光也很低。由于AIE效应，细胞摄取的TPE-TPP纳米聚集体在三光子激发下长时间耐光漂白。此外，我们证明，对于本TPE-TPP AIE的三光子显微镜（1020 nm激发）有一个更好的信噪比比双光子显微镜（810 nm激发）在组织成像。
Fluorescent probes are one of the most popularly used bioimaging markers to monitor metabolic processes of living cells. However, long-term light excitation always leads to photobleaching of fluorescent probes, unavoidable autofluorescence as well as photodamage of cells. To overcome these limitations, we synthesized a type of photostable luminogen named TPE-TPP with an aggregation induced emission (AIE) characteristic, and achieved its three-photon imaging with femtosecond laser excitation of 1020 nm. By using TPE-TPP as fluorescent probes, three-photon microscopy under 1020 nm excitation showed little photo-damage, as well as low autofluorescence to HeLa cells. Due to the AIE effect, the TPE-TPP nanoaggregates uptaken by cells were resistant to photobleaching under three-photon excitation for an extended period of time. Furthermore, we demonstrated that for the present TPE-TPP AIE the three-photon microscopy (with 1020 nm excitation) had a better signal to noise ratio than the two-photon microscopy (with 810 nm excitation) in tissue imaging.