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

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.

Intracellular Electrochemical Sensing

“…Finally, cell activity can be electrochemically detected by using marker proteins, such as endogenous alkaline phosphatase (ALP). For example, cell differentiation of embryonic stem cells and early‐stage bone differentiation have been electrochemically detected. Several types of integrated electrochemical devices have been developed for bioanalysis including cell analysis and they are discussed in detail elsewhere .…”

“…Further, electrode arrays are used for imaging and mapping of dopamine released from cells . Recently, a new electrochemical imaging approach based on electrode arrays, designated “electrochemicolor imaging” , was developed for simultaneous detection of multiple analytes, such as dopamine and dissolved oxygen. By using the imaging system, dopamine release and respiratory activity of neuron‐like cells were successfully imaged in real time (Figure ).…”

Advances and Perspectives in Chemical Imaging in Cellular Environments Using Electrochemical Methods

“…To enhance the ability of the sensor array device, the approach has been extended to offer simultaneous multi-reaction imaging. In electrochemicolor imaging, two (or more) different potentials are applied at alternate electrodes within the array (Figure 9) [136]. Alternate potentials can be applied at alternate electrodes on the device (V1 and V2 modes in Figure 9), with only moderate loss of imaging spatial resolution, using a mathematical approach to fill in the now "missing" pixels.…”

Simultaneous and Selective Imaging of Dopamine and Glutamate Using an Enzyme‐modified Large‐scale Integration (LSI)‐based Amperometric Electrochemical Device

“…These chip devices have also been used to characterize glutamatergic and dopaminergic cells and to examine brain slices as well. A new device was developed, Bio‐LSI , a large‐scale integration (LSI)‐based amperometric device containing 400 sensors with a pitch of 250 μm, and used to evaluate DA released from 3D‐cultured pheochromocytoma (PC12) cells .…”

“…An electrochemicolor imaging method which enables simultaneous imaging of multiple redox species, was applied for selective and simultaneous imaging of both neurotransmitters using the enzyme‐modified Bio‐LSI. Two electrochemical images reconstructed from 400 electrochemical signals from the DA and Glu electrodes were obtained using a mathematical approach, as described in the experimental section.…”