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1828 KiB

Open AccessArticle

Amperometric Microsensors Monitoring Glutamate-Evoked In Situ Responses of Nitric Oxide and Carbon Monoxide from Live Human Neuroblastoma Cells

by Yejin Ha, Chaejeong Heo, Juhyun Woo, Hyunwoo Ryu, Youngmi Lee and And Minah Suh

Sensors 2017, 17(7), 1661; https://doi.org/10.3390/s17071661 - 19 Jul 2017

Cited by 2 | Viewed by 4556

Abstract

In the brain, nitric oxide (NO) and carbon monoxide (CO) are important signaling gases which have multifaceted roles, such as neurotransmitters, neuromodulators, and vasodilators. Even though it is difficult to measure NO and CO in a living system due to their high diffusibility [...] Read more.

In the brain, nitric oxide (NO) and carbon monoxide (CO) are important signaling gases which have multifaceted roles, such as neurotransmitters, neuromodulators, and vasodilators. Even though it is difficult to measure NO and CO in a living system due to their high diffusibility and extremely low release levels, electrochemical sensors are promising tools to measure in vivo and in vitro NO and CO gases. In this paper, using amperometric dual and septuple NO/CO microsensors, real-time NO and CO changes evoked by glutamate were monitored simultaneously for human neuroblastoma (SH-SY5Y) cells. In cultures, the cells were differentiated and matured into functional neurons by retinoic acid and brain-derived neurotrophic factor. When glutamate was administrated to the cells, both NO and CO increases and subsequent decreases returning to the basal levels were observed with a dual NO/CO microsensor. In order to facilitate sensor’s measurement, a flower-type septuple NO/CO microsensor was newly developed and confirmed in terms of the sensitivity and selectivity. The septuple microsensor was employed for the measurements of NO and CO changes as a function of distances from the position of glutamate injection. Our sensor measurements revealed that only functionally differentiated cells responded to glutamate and released NO and CO. Full article

(This article belongs to the Special Issue Whole Cell-Based Biosensors and Application)

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11113 KiB

Open AccessArticle

Microfluidic Platform for the Long-Term On-Chip Cultivation of Mammalian Cells for Lab-On-A-Chip Applications

by Frank Bunge, Sander Van den Driesche and Michael J. Vellekoop

Sensors 2017, 17(7), 1603; https://doi.org/10.3390/s17071603 - 10 Jul 2017

Cited by 23 | Viewed by 5509

Abstract

Lab-on-a-Chip (LoC) applications for the long-term analysis of mammalian cells are still very rare due to the lack of convenient cell cultivation devices. The difficulties are the integration of suitable supply structures, the need of expensive equipment like an incubator and sophisticated pumps [...] Read more.

Lab-on-a-Chip (LoC) applications for the long-term analysis of mammalian cells are still very rare due to the lack of convenient cell cultivation devices. The difficulties are the integration of suitable supply structures, the need of expensive equipment like an incubator and sophisticated pumps as well as the choice of material. The presented device is made out of hard, but non-cytotoxic materials (silicon and glass) and contains two vertical arranged membranes out of hydrogel. The porous membranes are used to separate the culture chamber from two supply channels for gases and nutrients. The cells are fed continuously by diffusion through the membranes without the need of an incubator and low requirements on the supply of medium to the assembly. The diffusion of oxygen is modelled in order to find the optimal dimensions of the chamber. The chip is connected via 3D-printed holders to the macroscopic world. The holders are coated with Parlyene C to ensure that only biocompatible materials are in contact with the culture medium. The experiments with MDCK-cells show the successful seeding inside the chip, culturing and passaging. Consequently, the presented platform is a step towards Lab-on-a-Chip applications that require long-term cultivation of mammalian cells. Full article

(This article belongs to the Special Issue Whole Cell-Based Biosensors and Application)

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1282 KiB

Open AccessArticle

Tuning the Sensitivity of the PDR5 Promoter-Based Detection of Diclofenac in Yeast Biosensors

by Astrid Schuller, Gerhard Rödel and Kai Ostermann

Sensors 2017, 17(7), 1506; https://doi.org/10.3390/s17071506 - 26 Jun 2017

Cited by 7 | Viewed by 4417

Abstract

The commonly used drug diclofenac is an important environmental anthropogenic pollutant. Currently, detection of diclofenac is mainly based on chemical and physical methods. Here we describe a yeast biosensor that drives the diclofenac-dependent expression of a recombinant fluorescent protein from the authentic promoter [...] Read more.

The commonly used drug diclofenac is an important environmental anthropogenic pollutant. Currently, detection of diclofenac is mainly based on chemical and physical methods. Here we describe a yeast biosensor that drives the diclofenac-dependent expression of a recombinant fluorescent protein from the authentic promoter of the PDR5 gene. This key component of the pleiotropic drug response encodes a multidrug transporter that is involved in cellular detoxification. We analyse the effects on diclofenac sensitivity of artificial PDR5 promoter derivatives in wild-type and various yeast mutant strains. This approach enabled us to generate sensor strains with elevated drug sensitivity. Full article

(This article belongs to the Special Issue Whole Cell-Based Biosensors and Application)

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7331 KiB

Open AccessArticle

Study of the Relation between the Resonance Behavior of Thickness Shear Mode (TSM) Sensors and the Mechanical Characteristics of Biofilms

by Pedro Castro, Luis Elvira, Juan Ramón Maestre and Francisco Montero de Espinosa

Sensors 2017, 17(6), 1395; https://doi.org/10.3390/s17061395 - 15 Jun 2017

Cited by 7 | Viewed by 4346

Abstract

This work analyzes some key aspects of the behavior of sensors based on piezoelectric Thickness Shear Mode (TSM) resonators to study and monitor microbial biofilms. The operation of these sensors is based on the analysis of their resonance properties (both resonance frequency and [...] Read more.

This work analyzes some key aspects of the behavior of sensors based on piezoelectric Thickness Shear Mode (TSM) resonators to study and monitor microbial biofilms. The operation of these sensors is based on the analysis of their resonance properties (both resonance frequency and dissipation factor) that vary in contact with the analyzed sample. This work shows that different variations during the microorganism growth can be detected by the sensors and highlights which of these changes are indicative of biofilm formation. TSM sensors have been used to monitor in real time the development of Staphylococcus epidermidis and Escherichia coli biofilms, formed on the gold electrode of the quartz crystal resonators, without any coating. Strains with different ability to produce biofilm have been tested. It was shown that, once a first homogeneous adhesion of bacteria was produced on the substrate, the biofilm can be considered as a semi-infinite layer and the quartz sensor reflects only the viscoelastic properties of the region immediately adjacent to the resonator, not being sensitive to upper layers of the biofilm. The experiments allow the microrheological evaluation of the complex shear modulus (G* = G′ + jG″) of the biofilm at 5 MHz and at 15 MHz, showing that the characteristic parameter that indicates the adhesion of a biofilm for the case of S. epidermidis and E. coli, is an increase in the resonance frequency shift of the quartz crystal sensor, which is connected with an increase of the real shear modulus, related to the elasticity or stiffness of the layer. In addition both the real and the imaginary shear modulus are frequency dependent at these high frequencies in biofilms. Full article

(This article belongs to the Special Issue Whole Cell-Based Biosensors and Application)

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3901 KiB

Open AccessArticle

Probe Sensor Using Nanostructured Multi-Walled Carbon Nanotube Yarn for Selective and Sensitive Detection of Dopamine

by Wed Al-Graiti, Zhilian Yue, Javad Foroughi, Xu-Feng Huang, Gordon Wallace, Ray Baughman and Jun Chen

Sensors 2017, 17(4), 884; https://doi.org/10.3390/s17040884 - 18 Apr 2017

Cited by 37 | Viewed by 6657

Abstract

The demands for electrochemical sensor materials with high strength and durability in physiological conditions continue to grow and novel approaches are being enabled by the advent of new electromaterials and novel fabrication technologies. Herein, we demonstrate a probe-style electrochemical sensor using highly flexible [...] Read more.

The demands for electrochemical sensor materials with high strength and durability in physiological conditions continue to grow and novel approaches are being enabled by the advent of new electromaterials and novel fabrication technologies. Herein, we demonstrate a probe-style electrochemical sensor using highly flexible and conductive multi-walled carbon nanotubes (MWNT) yarns. The MWNT yarn-based sensors can be fabricated onto micro Pt-wire with a controlled diameter varying from 100 to 300 µm, and then further modified with Nafion via a dip-coating approach. The fabricated micro-sized sensors were characterized by electron microscopy, Raman, FTIR, electrical, and electrochemical measurements. For the first time, the MWNT/Nafion yarn-based probe sensors have been assembled and assessed for high-performance dopamine sensing, showing a significant improvement in both sensitivity and selectivity in dopamine detection in presence of ascorbic acid and uric acid. It offers the potential to be further developed as implantable probe sensors. Full article

(This article belongs to the Special Issue Whole Cell-Based Biosensors and Application)

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3560 KiB

Open AccessArticle

Electrical Impedance Monitoring of C2C12 Myoblast Differentiation on an Indium Tin Oxide Electrode

by Ilhwan Park, Yeonhee Hong, Young-Hoo Jun, Ga-Yeon Lee, Hee-Sook Jun, Jae-Chul Pyun, Jeong-Woo Choi and Sungbo Cho

Sensors 2016, 16(12), 2068; https://doi.org/10.3390/s16122068 - 5 Dec 2016

Cited by 14 | Viewed by 8739

Abstract

Electrical cell-substrate impedance sensing is increasingly being used for label-free and real-time monitoring of changes in cell morphology and number during cell growth, drug screening, and differentiation. In this study, we evaluated the feasibility of using ECIS to monitor C2C12 myoblast differentiation using [...] Read more.

Electrical cell-substrate impedance sensing is increasingly being used for label-free and real-time monitoring of changes in cell morphology and number during cell growth, drug screening, and differentiation. In this study, we evaluated the feasibility of using ECIS to monitor C2C12 myoblast differentiation using a fabricated indium tin oxide (ITO) electrode-based chip. C2C12 myoblast differentiation on the ITO electrode was validated based on decreases in the mRNA level of MyoD and increases in the mRNA levels of myogenin and myosin heavy chain (MHC). Additionally, MHC expression and morphological changes in myoblasts differentiated on the ITO electrode were comparable to those in cells in the control culture dish. From the monitoring the integration of the resistance change at 21.5 kHz, the cell differentiation was label-free and real-time detectable in 30 h of differentiation (p < 0.05). Full article

(This article belongs to the Special Issue Whole Cell-Based Biosensors and Application)

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Review

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3526 KiB

Open AccessReview

The Application of Whole Cell-Based Biosensors for Use in Environmental Analysis and in Medical Diagnostics

by Qingyuan Gui, Tom Lawson, Suyan Shan, Lu Yan and Yong Liu

Sensors 2017, 17(7), 1623; https://doi.org/10.3390/s17071623 - 13 Jul 2017

Cited by 242 | Viewed by 21244

Abstract

Various whole cell-based biosensors have been reported in the literature for the last 20 years and these reports have shown great potential for their use in the areas of pollution detection in environmental and in biomedical diagnostics. Unlike other reviews of this growing [...] Read more.

Various whole cell-based biosensors have been reported in the literature for the last 20 years and these reports have shown great potential for their use in the areas of pollution detection in environmental and in biomedical diagnostics. Unlike other reviews of this growing field, this mini-review argues that: (1) the selection of reporter genes and their regulatory proteins are directly linked to the performance of celllular biosensors; (2) broad enhancements in microelectronics and information technologies have also led to improvements in the performance of these sensors; (3) their future potential is most apparent in their use in the areas of medical diagnostics and in environmental monitoring; and (4) currently the most promising work is focused on the better integration of cellular sensors with nano and micro scaled integrated chips. With better integration it may become practical to see these cells used as (5) real-time portable devices for diagnostics at the bedside and for remote environmental toxin detection and this in situ application will make the technology commonplace and thus as unremarkable as other ubiquitous technologies. Full article

(This article belongs to the Special Issue Whole Cell-Based Biosensors and Application)

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2627 KiB

Open AccessReview

Bioluminescence Assays for Monitoring Chondrogenic Differentiation and Cartilage Regeneration

by Hyeon Jeong Je, Min Gu Kim and Hyuck Joon Kwon

Sensors 2017, 17(6), 1306; https://doi.org/10.3390/s17061306 - 6 Jun 2017

Cited by 8 | Viewed by 5954

Abstract

Since articular cartilage has a limited regeneration potential, for developing biological therapies for cartilage regeneration it is important to study the mechanisms underlying chondrogenesis of stem cells. Bioluminescence assays can visualize a wide range of biological phenomena such as gene expression, signaling, metabolism, [...] Read more.

Since articular cartilage has a limited regeneration potential, for developing biological therapies for cartilage regeneration it is important to study the mechanisms underlying chondrogenesis of stem cells. Bioluminescence assays can visualize a wide range of biological phenomena such as gene expression, signaling, metabolism, development, cellular movements, and molecular interactions by using visible light and thus contribute substantially to elucidation of their biological functions. This article gives a concise review to introduce basic principles of bioluminescence assays and applications of the technology to visualize the processes of chondrogenesis and cartilage regeneration. Applications of bioluminescence assays have been highlighted in the methods of real-time monitoring of gene expression and intracellular levels of biomolecules and noninvasive cell tracking within animal models. This review suggests that bioluminescence assays can be applied towards a visual understanding of chondrogenesis and cartilage regeneration. Full article

(This article belongs to the Special Issue Whole Cell-Based Biosensors and Application)

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8486 KiB

Open AccessReview

Cell Microarray Technologies for High-Throughput Cell-Based Biosensors

by Hye Jin Hong, Woong Sub Koom and Won-Gun Koh

Sensors 2017, 17(6), 1293; https://doi.org/10.3390/s17061293 - 5 Jun 2017

Cited by 38 | Viewed by 10319

Abstract

Due to the recent demand for high-throughput cellular assays, a lot of efforts have been made on miniaturization of cell-based biosensors by preparing cell microarrays. Various microfabrication technologies have been used to generate cell microarrays, where cells of different phenotypes are immobilized either [...] Read more.

Due to the recent demand for high-throughput cellular assays, a lot of efforts have been made on miniaturization of cell-based biosensors by preparing cell microarrays. Various microfabrication technologies have been used to generate cell microarrays, where cells of different phenotypes are immobilized either on a flat substrate (positional array) or on particles (solution or suspension array) to achieve multiplexed and high-throughput cell-based biosensing. After introducing the fabrication methods for preparation of the positional and suspension cell microarrays, this review discusses the applications of the cell microarray including toxicology, drug discovery and detection of toxic agents. Full article

(This article belongs to the Special Issue Whole Cell-Based Biosensors and Application)

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3580 KiB

Open AccessReview

Domain 4 (D4) of Perfringolysin O to Visualize Cholesterol in Cellular Membranes—The Update

by Masashi Maekawa

Sensors 2017, 17(3), 504; https://doi.org/10.3390/s17030504 - 3 Mar 2017

Cited by 42 | Viewed by 9191

Abstract

The cellular membrane of eukaryotes consists of phospholipids, sphingolipids, cholesterol and membrane proteins. Among them, cholesterol is crucial for various cellular events (e.g., signaling, viral/bacterial infection, and membrane trafficking) in addition to its essential role as an ingredient of steroid hormones, vitamin D, [...] Read more.

The cellular membrane of eukaryotes consists of phospholipids, sphingolipids, cholesterol and membrane proteins. Among them, cholesterol is crucial for various cellular events (e.g., signaling, viral/bacterial infection, and membrane trafficking) in addition to its essential role as an ingredient of steroid hormones, vitamin D, and bile acids. From a micro-perspective, at the plasma membrane, recent emerging evidence strongly suggests the existence of lipid nanodomains formed with cholesterol and phospholipids (e.g., sphingomyelin, phosphatidylserine). Thus, it is important to elucidate how cholesterol behaves in membranes and how the behavior of cholesterol is regulated at the molecular level. To elucidate the complexed characteristics of cholesterol in cellular membranes, a couple of useful biosensors that enable us to visualize cholesterol in cellular membranes have been recently developed by utilizing domain 4 (D4) of Perfringolysin O (PFO, theta toxin), a cholesterol-binding toxin. This review highlights the current progress on development of novel cholesterol biosensors that uncover new insights of cholesterol in cellular membranes. Full article

(This article belongs to the Special Issue Whole Cell-Based Biosensors and Application)

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