title: Electrochemiluminescence imaging for parallel single-cell analysis of active membrane cholesterol.
abstract: Luminol electrochemiluminescence (ECL) imaging was developed for the parallel measurement of active membrane cholesterol at single living cells, thus establishing a novel electrochemical detection technique for single cells with high analysis throughput and low detection limit. In our strategy, the luminescence generated from luminol and hydrogen peroxide upon the potential was recorded in one image so that hydrogen peroxide at the surface of multiple cells could be simultaneously analyzed. Compared with the classic microelectrode array for the parallel single-cell analysis, the plat electrode only was needed in our ECL imaging, avoiding the complexity of electrode fabrication. The optimized ECL imaging system showed that hydrogen peroxide as low as 10 μM was visible and the efflux of hydrogen peroxide from cells could be determined. Coupled with the reaction between active membrane cholesterol and cholesterol oxidase to generate hydrogen peroxide, active membrane cholesterol at cells on the electrode was analyzed at single-cell level. The luminescence intensity was correlated with the amount of active membrane cholesterol, validating our system for single-cell cholesterol analysis. The relative high standard deviation on the luminescence suggested high cellular heterogeneities on hydrogen peroxide efflux and active membrane cholesterol, which exhibited the significance of single-cell analysis. This success in ECL imaging for single-cell analysis opens a new field in the parallel measurement of surface molecules at single cells.
authors.affiliation: †School of Pharmacy, Nanjing Medical University, Jiangsu, 210000, China.
authors.forename: Junyu
authors.initials: J
authors.lastname: Zhou
authors.affiliation: ‡Key State Laboratory of Analytical Chemistry for Life Science and School of Chemistry and Chemical Engineering, Nanjing University, Jiangsu, 210093, China.
authors.forename: Guangzhong
authors.initials: G
authors.lastname: Ma
authors.affiliation: †School of Pharmacy, Nanjing Medical University, Jiangsu, 210000, China.
authors.forename: Yun
authors.initials: Y
authors.lastname: Chen
authors.affiliation: †School of Pharmacy, Nanjing Medical University, Jiangsu, 210000, China.
authors.forename: Danjun
authors.initials: D
authors.lastname: Fang
authors.affiliation: ‡Key State Laboratory of Analytical Chemistry for Life Science and School of Chemistry and Chemical Engineering, Nanjing University, Jiangsu, 210093, China.
authors.forename: Dechen
authors.initials: D
authors.lastname: Jiang
authors.affiliation: ‡Key State Laboratory of Analytical Chemistry for Life Science and School of Chemistry and Chemical Engineering, Nanjing University, Jiangsu, 210093, China.
authors.forename: Hong-Yuan
authors.initials: HY
authors.lastname: Chen
mesh_terms: D002784:Cholesterol / Q000032:analysis*; D055664:Electrochemical Techniques; D006367:HeLa Cells; D006801:Humans; D006861:Hydrogen Peroxide / Q000737:chemistry; D008163:Luminescent Measurements; D008566:Membranes / Q000737:chemistry; D059010:Single-Cell Analysis / Q000379:methods*
chemical_list: D002784:Cholesterol; D006861:Hydrogen Peroxide