fluorescein-thiosemicarbazide and metaperiodate

A simple and efficient method for selectively imaging and monitoring in situ the expression of sialylated glycoproteins on living cells has been developed. Treating living cells by mild periodate oxidation to selectively generate aldehydes on sialylated glycoproteins, followed by direct labeling of aldehydes with a commercially available fluorescent tag, fluorescein-5-thiosemicarbazide (FTSC), allows in situ imaging and quantification of sialylated glycoproteins on living cells. Under optimum reaction conditions, the periodate oxidation-based FTSC ligation (PF) strategy could be completed within 40 min. The cells undergoing the PF assay revealed a 91% viability and a fairly high-level of metabolic activity. Compared with current labeling methods, the PF assay proved to be a simpler and faster means of imaging sialylated glycoproteins on living cells. The PF assay has been successfully applied to imaging the location and quantification of the abundance of sialylated glycoproteins on tumor and normal cells. Our results demonstrated the methodological significance in clinical diagnosis and functional elucidation studies.

Topics: Cell Survival; Flow Cytometry; Fluoresceins; Fluorescent Dyes; Glycoproteins; HeLa Cells; Humans; Microscopy, Confocal; Microscopy, Fluorescence; Oxidants; Oxidation-Reduction; Periodic Acid; Sialic Acids; Staining and Labeling; Tissue Fixation