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- Title
Fluorometric visualization of mucin 1 glycans on cell surfaces based on rolling-mediated cascade amplification and CdTe quantum dots.
- Authors
Yang, XiaoTong; Tang, YingYing; Zhang, XiaoJing; Hu, Yue; Tang, Yu Ying; Hu, Lin Yu; Li, Su; Xie, Yaochen; Zhu, Dong
- Abstract
A rolling-mediated cascade (RMC) amplification strategy is described for improved visualization of profiling glycans of mucin 1 (MUC 1) on cell surfaces. CdTe quantum dots (QDs) are used as fluorescent labels. The RMC based amplification allows even distinct glycoforms of MUC1 to be visualized on the surface of MCF-7 cell via an amplified Förster resonance energy transfer (FRET) imaging strategy that works at excitation/emission wavelengths of 345/610 nm. This is achieved by utilizing antibody against MUC1 modified with the fluorescent label 7-amino-4-methylcoumarin-3-acetic acid (AMCA) as the energy donor in FRET. The QDs (used to label surface glycans) act as acceptors. N-Azidoacetylgalactosamine-Acetylated (Ac4GalNAz) as a non-natural azido sugar, can be incorporated into the glycans of the cell surface, which can promote further labeling. The method has the advantage of only requiring a small amount of non-natural sugar to be introduced in metabolic glycan labeling since too much of an artificial sugar will interfere with the physiological functions of cells.
- Subjects
CELL membranes; QUANTUM dots; GLYCANS; FLUORESCENCE resonance energy transfer; GLYCOCONJUGATES; VISUALIZATION
- Publication
Microchimica Acta, 2019, Vol 186, Issue 11, pN.PAG
- ISSN
0026-3672
- Publication type
Article
- DOI
10.1007/s00604-019-3840-8