Analysis of Intracellular Glucose at Single Cells Using Electrochemiluminescence Imaging.
Here, luminol electrochemiluminescence was first applied to analyze intracellular molecules, such as glucose, at single cells. The individual cells were retained in cell-sized microwells on a gold coated indium tin oxide (ITO) slide, which were treated with luminol, triton X-100, and glucose oxidase simultaneously. The broken cellular membrane in the presence of triton X-100 released intracellular glucose into the microwell and reacted with glucose oxidase to generate hydrogen peroxide, which induced luminol luminescence under positive potential. To achieve fast analysis, the luminescences from 64 individual cells on one ITO slide were imaged in 60 s using a charge-coupled device (CCD). More luminescence was observed at all the microwells after the introduction of triton X-100 and glucose oxidase suggested that intracellular glucose was detected at single cells. The starvation of cells to decrease intracellular glucose produced less luminescence, which confirmed that our luminescence intensity was correlated with the concentration of intracellular glucose. Large deviations in glucose concentration at observed single cells revealed high cellular heterogeneity in intracellular glucose for the first time. This developed electrochemiluminescence assay will be potentially applied for fast analysis of more intracellular molecules in single cells to elucidate cellular heterogeneity.
State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University , Nanjing, Jiangsu 210093, China.
Jingjing
J
Xu
State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University , Nanjing, Jiangsu 210093, China.
Peiyuan
P
Huang
State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University , Nanjing, Jiangsu 210093, China.
Yu
Y
Qin
State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University , Nanjing, Jiangsu 210093, China.
Dechen
D
Jiang
State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University , Nanjing, Jiangsu 210093, China.
Hong-Yuan
HY
Chen
D055664:Electrochemical Techniques; D005947:Glucose / Q000032:analysis*; D005949:Glucose Oxidase / Q000737:chemistry / Q000378:metabolism; D006046:Gold / Q000737:chemistry; D006801:Humans; D008163:Luminescent Measurements; D008165:Luminol / Q000737:chemistry; D008855:Microscopy, Electron, Scanning; D017830:Octoxynol / Q000737:chemistry; D061848:Optical Imaging; D059010:Single-Cell Analysis / Q000379:methods*; D017971:Tin Compounds / Q000737:chemistry
D017971:Tin Compounds; D008165:Luminol; C109984:indium tin oxide; D006046:Gold; D017830:Octoxynol; D005949:Glucose Oxidase; D005947:Glucose