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咔唑二阳离子盐在活体植物混浊组织中 DNA 的双光子荧光成像。

Two-photon fluorescence imaging of DNA in living plant turbid tissue with carbazole dicationic salt.

机构信息

State Key Laboratory of Crystal Materials, Shandong University, Jinan, 250100, China.

出版信息

Org Biomol Chem. 2010 Oct 21;8(20):4582-8. doi: 10.1039/c0ob00030b. Epub 2010 Aug 16.

Abstract

Three carbazole dicationic salts, namely 3,6-bis(1-methyl-4-vinylpyridinium) carbazole diiodide (BMVC), 9-ethyl-3,6-bis(1-methyl-4-vinylpyridinium) carbazole diiodide (9E-BMVC) and 9-ethyl-3,6-bis(1-hydroxyethyl-4-vinylpyridinium)carbazole diiodide (9E-BHVC), were synthesized successfully. Their photophysical properties were evaluated by absorption, one- and two-photon fluorescence spectra, and their higher fluorescence intensity and larger two-photon excited fluorescence action cross-sections (Φ×δ) in the presence of DNA than those in the absence of DNA give them good DNA two-photon light-switch properties. Furthermore, their ability to image nuclei in living plant cells and turbid tissues by using two-photon excited fluorescence was carefully studied, and the experimental results indicate that these dicationic salts can exclusively label nuclei in intact living plant cells and tissues. In particular, 9E-BHVC exhibits optimized DNA labeling performance. Very importantly, compared to DAPI, 9E-BHVC can be used to carry out deeper observation using the same incident power, or can be used to obtain usable fluorescent images by using a lower incident power.

摘要

三种咔唑二阳离子盐,即 3,6-双(1-甲基-4-乙烯基吡啶鎓)咔唑二碘化物(BMVC)、9-乙基-3,6-双(1-甲基-4-乙烯基吡啶鎓)咔唑二碘化物(9E-BMVC)和 9-乙基-3,6-双(1-羟乙基-4-乙烯基吡啶鎓)咔唑二碘化物(9E-BHVC),被成功合成。通过吸收光谱、单光子和双光子荧光光谱评估了它们的光物理性质,与不存在 DNA 时相比,它们在存在 DNA 时具有更高的荧光强度和更大的双光子激发荧光作用截面(Φ×δ),这使它们具有良好的 DNA 双光子光开关性质。此外,还仔细研究了它们通过双光子激发荧光对活植物细胞和混浊组织中的细胞核进行成像的能力,实验结果表明,这些二阳离子盐可以专一地标记完整活植物细胞和组织中的细胞核。特别是,9E-BHVC 表现出优化的 DNA 标记性能。非常重要的是,与 DAPI 相比,9E-BHVC 可以在相同的入射功率下进行更深层次的观察,或者可以在使用较低的入射功率时获得可用的荧光图像。

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