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Proc Natl Acad Sci U S A ; 113(41): 11436-11440, 2016 10 11.
Artigo em Inglês | MEDLINE | ID: mdl-27671654

RESUMO

The development of more intricate devices for the analysis of small molecules and protein activity in single cells would advance our knowledge of cellular heterogeneity and signaling cascades. Therefore, in this study, a nanokit was produced by filling a nanometer-sized capillary with a ring electrode at the tip with components from traditional kits, which could be egressed outside the capillary by electrochemical pumping. At the tip, femtoliter amounts of the kit components were reacted with the analyte to generate hydrogen peroxide for the electrochemical measurement by the ring electrode. Taking advantage of the nanotip and small volume injection, the nanokit was easily inserted into a single cell to determine the intracellular glucose levels and sphingomyelinase (SMase) activity, which had rarely been achieved. High cellular heterogeneities of these two molecules were observed, showing the significance of the nanokit. Compared with the current methods that use a complicated structural design or surface functionalization for the recognition of the analytes, the nanokit has adapted features of the well-established kits and integrated the kit components and detector in one nanometer-sized capillary, which provides a specific device to characterize the reactivity and concentrations of cellular compounds in single cells.


Assuntos
Técnicas Eletroquímicas/métodos , Nanotecnologia/métodos , Análise de Célula Única/métodos , Animais , Eletrodos , Glucose/análise , Células HeLa , Humanos , Camundongos , Modelos Teóricos , Esfingomielina Fosfodiesterase/metabolismo
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