Lanthanide Complex for Single-Molecule Fluorescent in Situ Hybridization and Background-Free Imaging

Su, Fei1; Chen, Shiyu2; Liu, Yuanhua2; Zhou, Jiajia1; Du, Zhongbo2; Luo, Xiongjian2; Wen, Shihui1,3; Jin, Dayong1,3

2024-03-06

10.1021/acs.analchem.3c04530

ANALYTICAL CHEMISTRY   影响因子和分区

0003-2700

1520-6882

Traditional single-molecule fluorescence in situ hybridization (smFISH) methods for RNA detection often face sensitivity challenges due to the low fluorescence intensity of the probe. Also, short-lived autofluorescence complicates obtaining clear signals from tissue sections. In response, we have developed an smFISH probe using highly grafted lanthanide complexes to address both concentration quenching and autofluorescence background. Our approach involves an oligo PCR incorporating azide-dUTP, enabling conjugation with lanthanide complexes. This method has proven to be stable, convenient, and cost-effective. Notably, for the mRNA detection in SKBR3 cells, the lanthanide probe group exhibited 2.5 times higher luminescence intensity and detected 3 times more signal points in cells compared with the Cy3 group. Furthermore, we successfully applied the probe to image HER2 mRNA molecules in breast cancer FFPE tissue sections, achieving a 2.7-fold improvement in sensitivity compared to Cy3-based probes. These results emphasize the potential of time-resolved smFISH as a highly sensitive method for nucleic acid detection, free of background fluorescence interference.

SCI ; EI

英语

其他

Australian Research Council["KQTD20170810110913065","20200925174735005"] ; Shenzhen Science and Technology Program["ACSRF658277","SQ2017YFGH001190"] ; Australia China Science and Research Fund Joint Research Centre for Point-of-Care Testing[DE220100846] ; Australian Research Council (ARC) Discovery Early Career Researcher Award Scheme[FL210100180] ; Australian Research Council[DE220100846]

Chemistry

Chemistry, Analytical

WOS:001180787800001

AMER CHEMICAL SOC

CHEMISTRY

Web of Science

1.Univ Technol Sydney, Inst Biomed Mat & Devices IBMD, Fac Sci, Sydney, NSW 2007, Australia
2.Southern Univ Sci & Technol, UTS SUStech Joint Res Ctr Biomed Mat & Devices, Dept Biomed Engn, Shenzhen 518055, Guangdong, Peoples R China
3.Eastern Inst Technol, Eastern Inst Adv Study, Ningbo 315200, Zhejiang, Peoples R China

Su, Fei,Chen, Shiyu,Liu, Yuanhua,et al. Lanthanide Complex for Single-Molecule Fluorescent in Situ Hybridization and Background-Free Imaging[J]. ANALYTICAL CHEMISTRY,2024,96(11).

Su, Fei.,Chen, Shiyu.,Liu, Yuanhua.,Zhou, Jiajia.,Du, Zhongbo.,...&Jin, Dayong.(2024).Lanthanide Complex for Single-Molecule Fluorescent in Situ Hybridization and Background-Free Imaging.ANALYTICAL CHEMISTRY,96(11).

Su, Fei,et al."Lanthanide Complex for Single-Molecule Fluorescent in Situ Hybridization and Background-Free Imaging".ANALYTICAL CHEMISTRY 96.11(2024).