In situ electrochemical imaging of membrane glycan expression on micropatterned adherent single cells.
A scanning electrochemical microscopic (SECM) method for in situ imaging of four types of membrane glycan motifs on single adherent cells was proposed using BGC-823 human gastric carcinoma (BGC) cells as the model. These adherent cells were first micropatterned in the microwell of poly(dimethylsiloxane) membrane for precisely controlling the localized surface interaction, and the membrane glycans were then specifically recognized with corresponding lectins labeled with horseradish peroxidase (HRP). On the basis of the enzymatic oxidization of ferrocenylmethanol (FMA) by H(2)O(2) to yield FMA(+), the glycan expression level was detected by the reduction current of FMA(+) at the SECM tip. The cell-surface glycans could, thus, be in situ imaged by SECM at a single-cell level without peeling the cells from culture dish. Under the optimized conditions, four types of membrane glycan motifs showed statistically distinguishable expression levels. The SECM results for different glycan motifs on adherent single cells were consistent with those estimated by flow cytometric assay. This work provides a reliable approach for in situ evaluation of the characteristic glycopattern of single living cells and can be applied in cell biologic study based on cell surface carbohydrate expression.
Xue
Yadong
Y
Key Laboratory of Analytical Chemistry for Life Science (Ministry of Education of China), Department of Chemistry, Nanjing University, Nanjing 210093, PR China.
Ding
Lin
L
Lei
Jianping
J
Yan
Feng
F
Ju
Huangxian
H
D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000737:chemistry* / Q000378:metabolism; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D005296:Ferrous Compounds / Q000737:chemistry; D005434:Flow Cytometry; D006735:Horseradish Peroxidase / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide / Q000737:chemistry; D020527:Microscopy, Scanning Probe / Q000379:methods*; D011134:Polysaccharides / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism
D004129:Dimethylpolysiloxanes; D005296:Ferrous Compounds; D011134:Polysaccharides; C071550:ferrocenemethanol; C013830:baysilon; D006861:Hydrogen Peroxide; D006735:Horseradish Peroxidase