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Tetrahedron Probes for Ultrasensitive In Situ Detection of Telomerase and Surface Glycoprotein Activity in Living Cells

机译:用于超细胞瘤的四面体探针对活细胞中端粒酶和表面糖蛋白活性的超声检测

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In the present study, a tetrahedron probe with encoded internal reference nanoparticles (NPs) was self-assembled by a complementary nucleic acid aptamer for simultaneous ratiometric detection of telomerase (TE) and epithelial cell-adhesion molecule (EpCAM) in living cells. In the presence of a target, the dissociation of gold (Au) NPs, which was modified with corresponding tags, resulted in decreased surface-enhanced Raman scattering (SEAS) signals. In addition, the ratios of Raman intensity at 1346 cm(-1)/1096 cm(-1) (TE) and 1614 cm-1/1096 cm(-1) (EpCAM) compared with the internal reference were demonstrated to quantify the level of TE and EpCAM, respectively, and can eliminate certain background noise. A good linear relationship was observed between them, and the linear range of TE and EpCAM in HeLa cells was 0.7 x 10(-12) to 37.5 x 10(-12) IU and 1.24 to 75.48 pg/mL with a limit of detection (LOD) of 7.6 x 10(-16) IU and 0.53 pg/mL, respectively, which were consistent with the results of Raman confocal imaging. Meanwhile, the versatility and specificity of the developed probes were confirmed in cell lines. These results provide a reliable and ultrasensitive strategy for the in situ detection of biomarkers and a new method for SERS-based tetrahedrons in the early diagnosis of cancer.
机译:在本研究中,具有编码的内参考纳米颗粒(NPS)的四面体探针通过互补核酸适体自组装,用于同时测量活细胞中的端粒酶(TE)和上皮细胞 - 粘附分子(EPCAM)。在靶的存在下,用相应的标签修饰的金(Au)NP的解离导致表面增强的拉曼散射(SEA)信号降低。另外,对1346cm(-1)/ 1096cm(-1)(TE)和1614cm-1 / 1096cm(-1)(EPCAM)的拉曼强度的比率与内部参考相比进行了证明,以量化TE和EPCAM的水平分别,可以消除某些背景噪音。在它们之间观察到良好的线性关系,HeLa细胞中TE和EPCAM的线性范围为0.7×10(-12)至37.5×10(-12)IU,具有检测限为1.24至75.48pg / ml( LOD分别为7.6×10(-16)IU和0.53pg / ml,与拉曼共焦成像的结果一致。同时,在细胞系中证实了发育探针的多功能性和特异性。这些结果提供了对生物标志物的原位检测的可靠和超细策略,以及在癌症早期诊断中的基于SERS的四边体的新方法。

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