Electrochemicolor Imaging Using an LSI-Based Device for Multiplexed Cell Assays.

Yusuke Kanno,Atsushi Suda,Ryota Kunikata,Hitoshi Shiku,Chika Sakamoto,Hiroya Abe,Tomokazu Matsue,Kumi Y Inoue,Takehiro Onodera,Kosuke Ino,Masahki Matsudaira

doi:10.1021/acs.analchem.7b03042

Yusuke Kanno, Atsushi Suda + Show 9 more

Open Access

https://doi.org/10.1021/acs.analchem.7b03042

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Abstract

Multiplexed bioimaging systems have triggered the development of effective assays, contributing new biological information. Although electrochemical imaging is beneficial for quantitative analysis in real time, monitoring multiple cell functions is difficult. We have developed a novel electrochemical imaging system, herein, using a large-scale integration (LSI)-based amperometric device for detecting multiple biomolecules simultaneously. This system is designated as an electrochemicolor imaging system in which the current signals from two different types of biomolecules are depicted as a multicolor electrochemical image. The mode-selectable function of the 400-electrode device enables the imaging system and two different potentials can be independently applied to the selected electrodes. The imaging system is successfully applied for detecting multiple cell functions of the embryonic stem (ES) cell and the rat pheochromocytoma (PC12) cell aggregates. To the best of our knowledge, this is the first time that a real-time electrochemical mapping technique for multiple electroactive species, simultaneously, has been reported. The imaging system is a promising bioanalytical method for exploring complex biological phenomena.

Highlights

B ioimaging is a powerful analytical method for bioanalysis to acquire visual and mapping information from single cells to three-dimensional cultured cells
We develop a novel electrochemical imaging system called the electrochemicolor imaging system for the simultaneous analysis of multiple biomolecules, beyond the conventional electrochemical imaging systems
Enzyme membranes consisting of glucose oxidase (GOx), alkaline phosphatase (ALP), GOx + ALP, and without an enzyme were introduced into solutions containing glucose as the enzymatic substrate for GOx, p-aminophenyl phosphate (PAPP) as the enzymatic substrate for ALP, or with no enzymatic substrate

Summary

Analytical Chemistry

On the oxidation of enzymatic products and have elucidated the differentiation levels of ES cells.[22,23] Amperometric devices can be used to determine the local O2 concentrations. Zhsanov et al reported that there was no O2 gradient in an extracellular medium containing differentiated PC12 cells with high K+ stimulation using a fluorescence-based sensing technique, which agrees well with our result.[38] research has revealed that O2 consumption decreases upon dopamine release on the surface of a slice of rat caudate nucleus, using fast-scan cyclic voltammetry (FSCV).[39] In electrochemical imaging with an electrode array, the electrochemical signals depend upon the position and a size of each cell aggregate.[40] more precise analysis regarding the relationship between the respiration activity and dopamine release is needed by a correction of the electrochemical signals, considering the position and size of each cell aggregate or by calculating the gradient of the oxygen concentration.[41]. The concept of electrochemicolor imaging is useful for multiple analysis

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■ ACKNOWLEDGMENTS

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