Applying DNA rolling circle amplification in fluorescence imaging of cell surface glycans labeled by a metabolic method.

Glycans on the cell surfaces are essential for cellular communication. Metabolically labeling glycans can introduce unnatural sugars into cellular glycans, which can facilitate further labeling. We report herein imaging cell surface glycosylation by using click chemistry and DNA rolling circle amplification (RCA) to improve detection sensitivity. Through the RCA amplification, the image resolution of a cell was significantly improved and much fewer unnatural sugars were used than required previously. The advantage of this method is that it avoids introducing too much unnatural sugar, which can interfere with normal, physiological cell function. Simultaneously, the enhanced fluorescence intensity conveniently facilitates the detection of cells' own biosynthetic glycans by simply using a microplate reader. The results indicate that the metabolically labelling ability is different for different carbohydrates and different cells. Next, the RCA technique was adopted in a fluorescence resonance energy transfer (FRET)-based methodology that facilitated the glycan imaging of specific proteins on the cell surface. This method is broadly applicable to imaging the glycosylation of cellular proteins. Our results highlight the applications of RCA in metabolic glycan labeling, which can be used to monitor the glycosylation status on cells, and study the means by which glycosylation regulates cell function.