Micro‐ and Nano‐Technologies for Lipid Bilayer‐Based Ion‐Channel Functional Assays

Hirano‐Iwata, Ayumi; Ishinari, Yutaka; Yamamoto, Hideaki; Niwano, Michio

doi:10.1002/asia.201403391

Cited by 30 publications

(32 citation statements)

References 75 publications

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“…6,7 More recently, mechanically stable bilayers have been be exploited, which can be formed, often semi-automatically, by virtue of advances in the bilayer formation technique. 8,9 Therefore, the design of transmembrane signaling associated with stable planar bilayer membranes will be interesting for constructing sophisticated biosensors that respond to a variety of targets.…”

Section: Introductionmentioning

confidence: 99%

A Planar Bilayer Lipid Membrane Sensor Using a Miniaturized Auto-patch System

et al. 2017

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Section: Introductionmentioning

confidence: 99%

A Planar Bilayer Lipid Membrane Sensor Using a Miniaturized Auto-patch System

et al. 2017

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“…Although the patchclamp method is considered to be the gold standard for evaluating channel activities, the method also has limitations in that the observed currents are dependent on the condition of the target cells (3). Therefore, the development of an alternate method for recording ion channel activities, complementing the patch-clamp method, would be highly desirable (5).…”

Section: Introductionmentioning

confidence: 99%

“…Reconstitution of ion channel proteins in artificial freestanding bilayer lipid membranes (BLMs) is another approach for recording ion channel activities (5,6). The BLM reconstitution system has the advantage that the researcher can control the composition of the system, including the solution phase and the lipid environment.…”

Section: Introductionmentioning

confidence: 99%

“…One of the most critical steps in the formation of channelembedded stable BLMs is the integration of ion channels into BLMs (5,15,16). Ion channel proteins of pharmaceutical interest are generally extracted from tissues or cell lines that overexpress the channels and are then reconstituted into lipid vesicles to form proteoliposomes or membrane vesicles.…”

Section: Introductionmentioning

confidence: 99%

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Reconstitution of Human Ion Channels into Solvent-free Lipid Bilayers Enhanced by Centrifugal Forces

Hirano‐Iwata

Ishinari

Yoshida

et al. 2016

Biophysical Journal

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Artificially formed bilayer lipid membranes (BLMs) provide well-defined systems for functional analyses of various membrane proteins, including ion channels. However, difficulties associated with the integration of membrane proteins into BLMs limit the experimental efficiency and usefulness of such BLM reconstitution systems. Here, we report on the use of centrifugation to more efficiently reconstitute human ion channels in solvent-free BLMs. The method improves the probability of membrane fusion. Membrane vesicles containing the human ether-a-go-go-related gene (hERG) channel, the human cardiac sodium channel (Na v 1.5), and the human GABA A receptor (GABA A R) channel were formed, and the functional reconstitution of the channels into BLMs via vesicle fusion was investigated. Ion channel currents were recorded in 67% of the BLMs that were centrifuged with membrane vesicles under appropriate centrifugal conditions (14-55 Â g). The characteristic channel properties were retained for hERG, Na v 1.5, and GABA A R channels after centrifugal incorporation into the BLMs. A comparison of the centrifugal force with reported values for the fusion force revealed that a centrifugal enhancement in vesicle fusion was attained, not by accelerating the fusion process but by accelerating the delivery of membrane vesicles to the surface of the BLMs, which led to an increase in the number of membrane vesicles that were available for fusion. Our method for enhancing the probability of vesicle fusion promises to dramatically increase the experimental efficiency of BLM reconstitution systems, leading to the realization of a BLM-based, high-throughput platform for functional assays of various membrane proteins.

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“…[8][9][10][11] Among them, electronic micro/nano sensors are particularly attractive owing to their high sensitivity response, miniaturized design, cost-effectiveness and speedy response. Field effect transistors (FETs) are surface affinity type sensors that can amplify the biological interactions into electrical signals, due to their high transconductance gain.…”

mentioning

confidence: 99%

A Comprehensive Model for Whole Cell Sensing and Transmembrane Potential Measurement Using FET Biosensors

Pulikkathodi

Sarangadharan

Chen

et al. 2018

ECS J. Solid State Sci. Technol.

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In this research, the development of a highly sensitive FET based biosensor technology for whole cell sensing and cellular transmembrane potential measurement is discussed and a quantitative model is proposed to elucidate the sensor mechanism. Electrical double layer (EDL) gated FET biosensor platform offers high sensitivity target analyte detection, even in high ionic strength solutions, thereby eliminating the need for complex sample pre-processing methods. Using this sensor platform, we have developed a whole cell sensing technology that can detect and count cells, and monitor cellular bioelectric signals such as transmembrane potential changes. A quantitative model is developed to explain the sensor mechanism and theoretical prediction shows good agreement with experimental results. The changes in the cellular transmembrane potential upon extracellular stimulus is modeled based on the EDL FET sensor response. Our investigation reveals that EDL FET biosensor platform can be used to study the bioelectric signals of cells, dynamically which is highly relevant to cell biology in applications of drug development, ion channel studies and disease model establishment. This sensor technology can also be used for point of care diagnostics, in the diagnosis and prognosis of diseases such as cancer. Since the discovery of cells, the basic building blocks of life, they have remained as crucial links to unraveling the pathophysiology of disease and understanding the complex, fundamental processes that sustain life. Several technologies are employed for the identification and the study of fundamental cell biology. Optical technologies are commonly used for most types of cell based experiments. Flow cytometry is the traditional choice where cells are flown in through a channel and optical microscopy is used to image, analyze and characterize cells.1,2 Often fluorescent probes are used to image specific molecules within the cell to either identify the type of cell or track particular cellular functions.3,4 Newer technologies such as surface plasmon resonance (SPR) and resonant waveguide grating (RWG) have been well established for cellular investigation.5-7 However, these technologies require sophisticated instrument and trained laboratory staff to perform the experiments, which significantly increases the associated cost and inconvenience. Also, long turnaround times, photobleaching of fluorescent probes and poor signal to noise ratio are some of the issues that need to be addressed to improve the efficiency and reliability of the optical technologies.Recently, several micro/nano technology based cellular diagnostic and analytical platforms have been developed to overcome the limitations of traditional optical based technologies. [8][9][10][11] Among them, electronic micro/nano sensors are particularly attractive owing to their high sensitivity response, miniaturized design, cost-effectiveness and speedy response. Field effect transistors (FETs) are surface affinity type sensors that can amplify the biological interactions ...

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Micro‐ and Nano‐Technologies for Lipid Bilayer‐Based Ion‐Channel Functional Assays

Cited by 30 publications

References 75 publications

A Planar Bilayer Lipid Membrane Sensor Using a Miniaturized Auto-patch System

A Planar Bilayer Lipid Membrane Sensor Using a Miniaturized Auto-patch System

Reconstitution of Human Ion Channels into Solvent-free Lipid Bilayers Enhanced by Centrifugal Forces

A Comprehensive Model for Whole Cell Sensing and Transmembrane Potential Measurement Using FET Biosensors

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