Ho Lin-Chen, Wu Wei-Cheng, Chang Chang-Yu, Hsieh Hao-Hsuan, Lee Ching-Hsiao, Chang Huan-Tsung
†Department of Chemistry, National Taiwan University, 1, Section 4, Roosevelt Road, Taipei, Taiwan.
‡Nano Science and Technology Program, Taiwan International Graduate Program, Academia Sinica, Taipei, Taiwan and National Tsing-Hua University, Hsinchu, Taiwan.
Anal Chem. 2015;87(9):4925-32. doi: 10.1021/acs.analchem.5b00569. Epub 2015 Apr 21.
We have developed a simple, sensitive, and rapid fluorescence assay for the detection of cancer cells, based on "turn-on" retro-self-quenched fluorescence inside the cells. 1,3-Phenylenediamine resin (DAR) nanoparticles (NPs) containing rhodamine 6G (R6G) are conjugated with aptamer (apt) sgc8c to prepare sgc8c-R6GDAR NPs, while that containing rhodamine 101 (R101) are conjugated with TD05 for the preparation of TD05-R101DAR NPs. The sgc8c-R6GDAR and TD05-R101DAR NPs separately recognize CCRF-CEM and Ramos cells. The fluorescence intensities of the two apt-DAR NPs are both weak due to self-quenching, but they increase inside the cells as a result of release of the fluorophores from the apt-DAR NPs. The apt-DAR NPs' structure becomes less compact at low pH value, leading to the release of the fluorophores. The sgc8c-R6GDAR and TD05-R101DAR NPs allow detection of as low as 44 CCRF-CEM cells and 79 Ramos cells mL(-1), respectively, using a commercial reader within 10 min. Practicality of the two probes have been validated by the quantitation and identification of CCRF-CEM and Ramos cells spiked in blood samples through conventional fluorescence and flow cytometry analysis, with advantages of sensitivity, selectivity, and rapidity.
我们基于细胞内“开启”的反向自猝灭荧光,开发了一种用于检测癌细胞的简单、灵敏且快速的荧光检测方法。含有罗丹明6G(R6G)的1,3 - 苯二胺树脂(DAR)纳米颗粒(NPs)与适配体(apt)sgc8c偶联,制备sgc8c - R6GDAR NPs,而含有罗丹明101(R101)的则与TD05偶联,用于制备TD05 - R101DAR NPs。sgc8c - R6GDAR和TD05 - R101DAR NPs分别识别CCRF - CEM和Ramos细胞。由于自猝灭,两种适配体 - DAR NPs的荧光强度都较弱,但由于荧光团从适配体 - DAR NPs中释放,它们在细胞内的荧光强度会增加。在低pH值下,适配体 - DAR NPs的结构变得不那么紧密,导致荧光团释放。使用商用读数仪,sgc8c - R6GDAR和TD05 - R101DAR NPs分别能够在10分钟内检测低至44个CCRF - CEM细胞和79个Ramos细胞mL(-1)。通过传统荧光和流式细胞术分析对掺入血样中的CCRF - CEM和Ramos细胞进行定量和鉴定,验证了这两种探针的实用性,具有灵敏度高(原文sensitivity误写成sensitity)、选择性好和快速的优点。
需注意,原文中sensitity拼写错误,正确拼写为sensitivity 。