Jia Ti, Fu Congying, Huang Chusen, Yang Haotian, Jia Nengqin
The Education Ministry Key Laboratory of Resource Chemistry and Shanghai Key Laboratory of Rare Earth Functional Materials, Department of Chemistry, College of Life and Environmental Sciences, Shanghai Normal University, 100 Guilin Road, Shanghai 200234, China.
ACS Appl Mater Interfaces. 2015 May 13;7(18):10013-21. doi: 10.1021/acsami.5b02429. Epub 2015 May 1.
Fluorescence polarization (FP)-based signal is a self-referencing fluorescence signal, and it is less dependent on dye concentration and environmental interferences, which makes FP measurement an attractive alternative sensing technology to fluorescence intensity-based detection. However, most of the fluorescence polarization probes were constructed by introducing fluorescein, rhodamine, and cyanine dyes, which have relatively shorter excited-state lifetimes compared with BODIPY and naphthalimide dyes. Herein, a first naphthalimide based fluorescence polarization probe (BIO) was designed and synthesized for selective and direct detection of cancer cells. The relatively longer excited-state lifetimes and high photostability of naphthalimide makes BIO more sensitive and accuracy in quantitative determination of HeLa cells in homogeneous solution without cell lysis and further separation steps. The detection limit of BIO for HeLa cells was about 85 cells mL(-1), the linear range was from 2.5 × 10(2) cells mL(-1) to 1 × 10(6) cells mL(-1) and the response time is no more than 25 min. Moreover, due to the relatively high photostability of naphthalimide, BIO was particularly suitable for live cell imaging under continuous irradiation with confocal microscopy, and the specific interaction of BIO with CD44-overexpressing cell lines was clearly visualized. Importantly, this BIO based sensing platform offers a direct and real-time tool for cancer cell diagnosis when complemented with the use of naphthalimide-based fluorescence polarization probe.
基于荧光偏振(FP)的信号是一种自参考荧光信号,它对染料浓度和环境干扰的依赖性较小,这使得FP测量成为基于荧光强度检测的一种有吸引力的替代传感技术。然而,大多数荧光偏振探针是通过引入荧光素、罗丹明和花青染料构建的,与硼二吡咯和萘二甲酰亚胺染料相比,它们的激发态寿命相对较短。在此,设计并合成了一种基于萘二甲酰亚胺的新型荧光偏振探针(BIO),用于选择性直接检测癌细胞。萘二甲酰亚胺相对较长的激发态寿命和高光稳定性使得BIO在无需细胞裂解和进一步分离步骤的情况下,能够更灵敏、准确地定量测定均相溶液中的HeLa细胞。BIO对HeLa细胞的检测限约为85个细胞/mL,线性范围为2.5×10²个细胞/mL至1×10⁶个细胞/mL,响应时间不超过25分钟。此外,由于萘二甲酰亚胺具有较高的光稳定性,BIO特别适合在共聚焦显微镜连续照射下进行活细胞成像,并且能够清晰地观察到BIO与过表达CD44的细胞系之间的特异性相互作用。重要的是,当与基于萘二甲酰亚胺的荧光偏振探针配合使用时,这种基于BIO的传感平台为癌细胞诊断提供了一种直接实时的工具。