Development of a carboxyl-terminated indium tin oxide electrode for improving cell adhesion and facilitating low noise, real-time impedance measurements.

The working electrode's surface property is crucial to cell adhesion and signal collection in electric cell-substrate impedance sensing (ECIS). To date, the indium tin oxide (ITO)-based working electrode is of interest in ECIS study due to its high transparency and biocompatibility. Of great concern is the impedance signal loss, distortion, and data interpretation conflict profoundly created by the movement of multiple cells during ECIS study. Here, a carboxyl-terminated ITO substrate was prepared by stepwise surface amino silanization, with N-hydroxy succinimide (NHS) and 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide hydrochloride (EDC) treatment, respectively. We investigated the stepwise changes in the property of the treated ITO, cell-substrate adhesion, collective cell mobility, and time course of change in absolute impedance from multiple Chinese hamster ovary (CHO) cells [(Δt-Δ|Z|)CELLS]. The carboxyl-terminated ITO substrate with a surface roughness of 6.37 nm shows enhanced conductivity, 75% visible light transparency, improved cell adherence, reduced collective cell migration speed by approximately twofold, and diminished signal distortion in the [(Δt-Δ|Z|)CELLS]. Thus, our study provides an ITO surface-treatment strategy to reduce multiple cell movement effects and to obtain essential cell information from the ECIS study of multiple cells through undistorted (Δt-Δ|Z|)CELLS.

Portable molecularly imprinted polymer-based platform for detection of histamine in aqueous solutions.

Histamine, which is a naturally occurring chemical in seafood, is known to cause undesirable inflammatory response when consumed in large amounts. Histamine is produced in unsafe amounts in colored seafood when improperly stored for just a few hours. Food and health regulatory bodies across the world have guidelines limiting the amount of histamine in fresh as well as processed seafood. Conventional histamine detection is performed in testing labs, which is a slow process and results in bottlenecks in the seafood supply-chain system. A system to rapidly detect the seafood histamine levels on site is very desirable for seafood suppliers. Herein, we describe an impedance-based histamine detection sensor built on a flexible substrate that can detect histamine in the range of 100-500 ppm. Moreover, our sensor discriminates histamine in the presence of DL-histidine and other biogenic amines, with the selectivity provided by molecular imprinting technology. As a proof of concept, a smartphone controlled, portable semi-quantitative histamine sensing device was fabricated that gave out reliable testing results for histamine in different test solutions as well as for real seafood. We believe this technology can be extended towards determination of other food contaminants in aqueous solutions.

Bioinspired, Self-Powered, and Highly Sensitive Electronic Skin for Sensing Static and Dynamic Pressures.

Flexible piezoresistive pressure sensors obtain global research interest owing to their potential applications in healthcare, human-robot interaction, and artificial nerves. However, an additional power supply is usually required to drive the sensors, which results in increased complexity of the pressure sensing system. Despite the great efforts in pursuing self-powered pressure sensors, most of the self-powered devices can merely detect the dynamic pressure and the reliable static pressure detection is still challenging. With the help of redox-induced electricity, a bioinspired graphite/polydimethylsiloxane piezoresistive composite film acting both as the cathode and pressure sensing layer, a neoteric electronic skin sensor is presented here to detect not only the dynamic forces but also the static forces without an external power supply. Additionally, the sensor exhibits a fascinating pressure sensitivity of ∼103 kPa-1 over a broad sensing range from 0.02 to 30 kPa. Benefiting from the advanced performance of the device, various potential applications including arterial pulse monitoring, human motion detecting, and Morse code generation are successfully demonstrated. This new strategy could pave a way for the development of next-generation self-powered wearable devices.

Gating a Single Cell: A Label-Free and Real-Time Measurement Method for Cellular Progression.

There is an ever-growing need for more advanced methods to study the response of cancer cells to new therapies. To determine cancer cells' response from a cell-mortality perspective to various cancer therapies, we report a label-free and real time method to monitor the in situ response of individual HeLa cells using a single cell gated transistor (SCGT). As a cell undergoes apoptotic cell death, it experiences changes in morphology and ion concentrations. This change is well in line with the threshold voltage of the SCGT, which has been verified by correlating the data with the cell morphologies by scanning electron microscopy and the ion-concentration analysis by inductively-coupled plasma mass spectrometry (ICPMS). This SCGT could replace patch clamps to study single cell activity via direct measurement in real time. Importantly, this SCGT can be used to study the electrical response of a single cell to stimuli that leaves the membrane intact.

Development of a Visible Light Triggerable Traceless Staudinger Ligation Reagent.

A series of substituted 9-methylenylanthracene photocages for diphenylphosphinothioesters have been synthesized to explore their photo-uncaging properties by visible light. Substituents such as phenyl, p-trifluoromethylphenyl, p-methoxyphenyl, ethyn-1-ylbenzene, and 3,3-dimethylbut-1-yn-1-yl have been introduced in order to extend the π-conjugation of the photocage and to shift the wavelength response of the uncaging process to the visible spectral range. Among these new photocages, the (10-(3,3-dimethylbut-1-yn-1-yl)anthracen-9-yl)methyl has been shown to have the best performance in terms of fast photo-uncaging and minimal byproduct formation. It is responsive to both UV and visible photoexcitation. Quantum yields of the photoinduced heterolytic anthracenylmethyl-phosphorus bond cleavage at 366 and 416 nm were found to be 0.08 and 0.025, respectively. This photocage enables traceless Staudinger ligation to be triggered by photoirradiation in the visible spectral range for bioconjugation applications. We demonstrate this with a series of visible-light-induced oligopeptide syntheses via the conjugation of amino acid/oligopeptide building blocks by the characteristic peptide linkage attained by traceless Staudinger ligation. Yields of the resultant conjugated oligopeptides ranged from 31 to 43%. This new photocage opens up the possibility of in situ synthesis of functional proteins/peptides mediated by visible-light-induced photoclick processes for the regulation of cellular/metabolic functions of life systems.

The molecularly imprinted polymer essentials: curation of anticancer, ophthalmic, and projected gene therapy drug delivery systems.

The development of polymeric materials as drug delivery systems has advanced from systems that rely on classical passive targeting to carriers that can sustain the precisely controlled release of payloads upon physicochemical triggers in desired microenvironment. Molecularly imprinted polymers (MIP), materials designed to capture specific molecules based on their molecular shape and charge distribution, are attractive candidates for fulfilling these purposes. In particular, drug-imprinted polymers coupled with active targeting mechanisms have been explored as potential drug delivery systems. In this review, we have curated important recent efforts in the development of drug-imprinted polymers in a variety of clinical applications, especially oncology and ophthalmology. MIP possesses properties that may complement the traditional delivery systems of these two disciplines, such as passive enhanced permeability and retention effect (EPR) in cancer tumors, and passive drug diffusion in delivering ophthalmic therapeutics. Furthermore, the prospects of MIP integration with the emerging gene therapies will be discussed.

Dual-Gated Transistor Platform for On-Site Detection of Lead Ions at Trace Levels.

On-site monitoring of heavy metals in drinking water has become crucial because of several high profile instances of contamination. Presently, reliable techniques for trace level heavy metal detection are mostly laboratory based, while the detection limits of contemporary field-based methods are barely meeting the exposure limits set by regulatory bodies such as the World Health Organization (WHO). Here, we show an on-site deployable, Pb2+ sensor on a dual-gated transistor platform whose lower detection limit is 2 orders of magnitude better than the traditional sensor and 1 order of magnitude lower than the exposure limit set by WHO. The enhanced sensitivity of our design is verified by numerically solving PNP (Planck-Nernst-Poisson) model. We demonstrate that the enhanced sensitivity is due to the suppression of ionic flux. The simplicity and the robustness of the design make it applicable for on-site screening, thereby facilitating rapid response to contamination events.

Profiling of Selected Functional Metabolites in the Central Nervous System of Marine Medaka (Oryzias melastigma) for Environmental Neurotoxicological Assessments.

The simultaneous profiling of 43 functional metabolites in the brain of the small model vertebrate organism, marine medaka (Oryzais melastigma), has been accomplished via dansyl chloride derivatization and LC-MS/MS quantification. This technique was applied to examine effects of 2,2',4,4'-tetrabromodiphenyl ether (BDE-47), one of the most abundant polybrominated diphenyl ether flame retardants in the natural environment, on the central nervous system (CNS) of vertebrates. The model teleosts were fed with bioencapsulated Artemia nauplii for up to 21 days. Multivariate statistical analysis has demonstrated that levels of numerous classical neurotransmitters and their metabolites in the CNS of the fish were perturbed even at the early phase of dietary exposure. Subsequent metabolic pathway analysis further implied potential impairment of the arginine and proline metabolism; glycine, serine and threonine metabolism; D-glutamine and D-glutamate metabolism; alanine, aspartate, and glutamate metabolism; valine, leucine, and isoleucine biosynthesis, and the cysteine and methionine metabolism in the brain of the test organism. Our results demonstrate that targeted profiling of functional metabolites in the CNS may shed light on how the various neurological pathways of vertebrates, including humans, are affected by toxicant/stress exposure.

A Photo-Triggered Traceless Staudinger-Bertozzi Ligation Reaction.

The use of light to control the course of a chemical/biochemical reaction is an attractive idea because of its ease of administration with high precision and fine spatial resolution. Staudinger ligation is one of the commonly adopted conjugation processes that involve a spontaneous reaction between azides and arylphosphines to form iminophosphoranes, which further hydrolyze to give stable amides. We designed an anthracenylmethyl diphenylphosphinothioester (1) that showed promising Staudinger ligation reactivity upon photo-excitation. Broadband photolysis at 360-400 nm in aqueous organic solvents induced heterolytic cleavage of its anthracenylmethyl-phosphorus bond, releasing a diphenylphosphinothioester (2) as an efficient traceless Staudinger-Bertozzi ligation reagent. The quantum yield of such a photo-induced heterolytic bond-cleavage at the optimal wavelength of photolysis (376 nm) at room temperature is ≥0.07. This work demonstrated the feasibility of photocaging arylphosphines to realize the photo-triggering of the Staudinger ligation reaction.

Ratiometric fluorescent probe for enantioselective detection of D-cysteine in aqueous solution.

A ratiometric fluorescent probe based on a Cd(2+)-ACAQ complex was designed and demonstrated for the chemo- and enantioselective detection of cysteine in 99:1 buffered HEPES:ACN solutions. Under the measuring conditions, the sensor demonstrates high selectivity toward Cys against Hcy and GSH, and an enantioselectivity of 3.35 can be achieved for antipodal forms of Cys.