Real-time gas mass spectroscopy by multivariate analysis.

Early and significant results for a real-time, column-free miniaturized gas mass spectrometer in detecting target species with partial overlapping spectra are reported. The achievements have been made using both nanoscale holes as a nanofluidic sampling inlet system and a robust statistical technique. Even if the presented physical implementation could be used with gas chromatography columns, the aim of high miniaturization requires investigating its detection performance with no aid. As a study case, in the first experiment, dichloromethane (CH2Cl2) and cyclohexane (C6H12) with concentrations in the 6-93 ppm range in single and compound mixtures were used. The nano-orifice column-free approach acquired raw spectra in 60 s with correlation coefficients of 0.525 and 0.578 to the NIST reference database, respectively. Then, we built a calibration dataset on 320 raw spectra of 10 known different blends of these two compounds using partial least square regression (PLSR) for statistical data inference. The model showed a normalized full-scale root-mean-square deviation (NRMSD) accuracy of [Formula: see text] and [Formula: see text] for each species, respectively, even in combined mixtures. A second experiment was conducted on mixes containing two other gasses, Xylene and Limonene, acting as interferents. Further 256 spectra were acquired on 8 new mixes, from which two models were developed to predict CH2Cl2 and C6H12, obtaining NRMSD values of 6.4% and 13.9%, respectively.

Development of an electrical impedance tomography set-up for the quantification of mineralization in biopolymer scaffolds.

Objective. 3D cell cultures are becoming a fundamental resource forin-vitrostudies, as they mimic more closelyin-vivobehavior. The analysis of these constructs, however, generally rely on destructive techniques, that prevent the monitoring over time of the same construct, thus increasing the results variability and the resources needed for each experiment.Approach. In this work, we focus on mineralization, a crucial process during maturation of artificial bone models, and propose electrical impedance tomography (EIT) as an alternative non-destructive approach. In particular, we discuss the development of an integrated hardware/software system capable of acquiring experimental data from 3D scaffolds and reconstructing the corresponding conductivity maps. We also show how the same software can test how the measurement is affected by biological features such as scaffold shrinking during the culture.Main results. An initial validation, comprising the acquisition of both a non-conductive phantom and alginate/gelatin scaffolds with known calcium content will be presented, together with thein-silicostudy of a cell-induced mineralization process. This analysis will allow for an initial verification of the systems functionality while limiting the effects of biological variability due to cell number and activity.Significance. Our results show the potential of EIT for the non-destructive quantification of matrix mineralization in 3D scaffolds, and open to the possible long term monitoring of this fundamental hallmark of osteogenic differentiation in hybrid tissue engineered constructs.

Sensing Technology for Fish Freshness and Safety: A Review.

Standard analytical methods for fish freshness assessment are based on the measurement of chemical and physical attributes related to fish appearance, color, meat elasticity or texture, odor, and taste. These methods have plenty of disadvantages, such as being destructive, expensive, and time consuming. All these techniques require highly skilled operators. In the last decade, rapid advances in the development of novel techniques for evaluating food quality attributes have led to the development of non-invasive and non-destructive instrumental techniques, such as biosensors, e-sensors, and spectroscopic methods. The available scientific reports demonstrate that all these new techniques provide a great deal of information with only one test, making them suitable for on-line and/or at-line process control. Moreover, these techniques often require little or no sample preparation and allow sample destruction to be avoided.

A Non-Invasive Soil Moisture Sensing System Electronic Architecture: A Real Environment Assessment.

This paper will show the electronic architecture of a portable and non-invasive soil moisture system based on an open rectangular waveguide. The spectral information, measured in the range of 1.5-2.7 GHz, is elaborated on by an embedded predictive model, based on a partial least squares (PLS) regression tool, for the estimation of the soil moisture (%) in a real environment. The proposed system is composed of a waveguide, containing Tx and Rx antennas, and an electronic circuit driven by a microcontroller (MCU). It will be shown how the system provides a useful and fast estimation of moisture on a silty clay loam soil characterized by a moisture range of about 9% to 32% and a soil temperature ranging from about 8 °C and 18 °C. Using the PLS approach, the moisture content can be predicted with an R2 value of 0.892, a root mean square error (RMSE) of 1.0%, and a residual prediction deviation (RPD) of 4.3. The results prove that it is possible to make accurate and rapid moisture assessments without the use of invasive electrodes, as currently employed by state-of-the-art approaches.

Optimum Design Rules for CMOS Hall Sensors.

This manuscript analyzes the effects of design parameters, such as aspect ratio, doping concentration and bias, on the performance of a general CMOS Hall sensor, with insight on current-related sensitivity, power consumption, and bandwidth. The article focuses on rectangular-shaped Hall probes since this is the most general geometry leading to shape-independent results. The devices are analyzed by means of 3D-TCAD simulations embedding galvanomagnetic transport model, which takes into account the Lorentz force acting on carriers due to a magnetic field. Simulation results define a set of trade-offs and design rules that can be used by electronic designers to conceive their own Hall probes.

A Low-Noise Transimpedance Amplifier for BLM-Based Ion Channel Recording.

High-throughput screening (HTS) using ion channel recording is a powerful drug discovery technique in pharmacology. Ion channel recording with planar bilayer lipid membranes (BLM) is scalable and has very high sensitivity. A HTS system based on BLM ion channel recording faces three main challenges: (i) design of scalable microfluidic devices; (ii) design of compact ultra-low-noise transimpedance amplifiers able to detect currents in the pA range with bandwidth >10 kHz; (iii) design of compact, robust and scalable systems that integrate these two elements. This paper presents a low-noise transimpedance amplifier with integrated A/D conversion realized in CMOS 0.35 µm technology. The CMOS amplifier acquires currents in the range ±200 pA and ±20 nA, with 100 kHz bandwidth while dissipating 41 mW. An integrated digital offset compensation loop balances any voltage offsets from Ag/AgCl electrodes. The measured open-input input-referred noise current is as low as 4 fA/√Hz at ±200 pA range. The current amplifier is embedded in an integrated platform, together with a microfluidic device, for current recording from ion channels. Gramicidin-A, α-haemolysin and KcsA potassium channels have been used to prove both the platform and the current-to-digital converter.

AC and Phase Sensing of Nanowires for Biosensing.

Silicon nanowires are label-free sensors that allow real-time measurements. They are economical and pave the road for point-of-care applications but require complex readout and skilled personnel. We propose a new model and technique for sensing nanowire sensors using alternating currents (AC) to capture both magnitude and phase information from the sensor. This approach combines the advantages of complex impedance spectroscopy with the noise reduction performances of lock-in techniques. Experimental results show how modifications of the sensors with different surface chemistries lead to the same direct-current (DC) response but can be discerned using the AC approach.

A Distributed Amplifier System for Bilayer Lipid Membrane (BLM) Arrays With Noise and Individual Offset Cancellation.

Lipid bilayer membrane (BLM) arrays are required for high throughput analysis, for example drug screening or advanced DNA sequencing. Complex microfluidic devices are being developed but these are restricted in terms of array size and structure or have integrated electronic sensing with limited noise performance. We present a compact and scalable multichannel electrophysiology platform based on a hybrid approach that combines integrated state-of-the-art microelectronics with low-cost disposable fluidics providing a platform for high-quality parallel single ion channel recording. Specifically, we have developed a new integrated circuit amplifier based on a novel noise cancellation scheme that eliminates flicker noise derived from devices under test and amplifiers. The system is demonstrated through the simultaneous recording of ion channel activity from eight bilayer membranes. The platform is scalable and could be extended to much larger array sizes, limited only by electronic data decimation and communication capabilities.

Noise limits of CMOS current interfaces for biosensors: a review.

Current sensing readout is one of the most frequent techniques used in biosensing due to the charge-transfer phenomena occurring at solid-liquid interfaces. The development of novel nanodevices for biosensing determines new challenges for electronic interface design based on current sensing, especially when compact and efficient arrays need to be organized, such as in recent trends of rapid label-free electronic detection of DNA synthesis. This paper will review the basic noise limitations of current sensing interfaces with particular emphasis on integrated CMOS technology. Starting from the basic theory, the paper presents, investigates and compares charge-sensitive amplifier architectures used in both continuous-time and discrete-time approaches, along with their design trade-offs involving noise floor, sensitivity to stray capacitance and bandwidth. The ultimate goal of this review is providing analog designers with helpful design rules and analytical tools. Also, in order to present a comprehensive overview of the state-of-the-art, the most relevant papers recently appeared in the literature about this topic are discussed and compared.

Lysis-on-chip of single target cells following forced interaction with CTLs or NK cells on a dielectrophoresis-based array.

Guiding the interaction of single cells acting as partners in heterotypic interactions (e.g., effectors and targets of immune lysis) and monitoring the outcome of these interactions are regarded as crucial biomedical achievements. In this study, taking advantage of a dielectrophoresis (DEP)-based Laboratory-on-a-chip platform (the DEPArray), we show that it is possible to generate closed DEP cages entrapping CTLs and NK cells as either single cells or clusters; reversibly immobilize a single virus-presenting or tumor cell within the chip at a selected position; move cages and their content to predetermined spatial coordinates by software-guided routing; force a cytotoxic effector to physically interact with a putative target within a secluded area by merging their respective cages; generate cages containing effector and target cells at predetermined E:T ratios; accurately assess cytotoxicity by real-time quantitation of the release kinetics of the fluorescent dye calcein from target cells (>50 lytic events may be tested simultaneously); estimate end points of calcein release within 16 min of initial E:T cell contact; simultaneously deliver Ab-based phenotyping and on-chip lysis assessment; and identify lytic and nonlytic E:T combinations and discriminate nonlytic effector phenotypes from target refractoriness to immune lysis. The proof of principle is provided that DEPArray technology, previously used to levitate and move single cells, can be used to identify highly lytic antiviral CTLs and tumor cells that are particularly refractory to NK cell lysis. These findings are of primary interest in targeted immunotherapy.

Programmable interactions of functionalized single bioparticles in a dielectrophoresis-based microarray chip.

Manipulating single biological objects is a major unmet challenge of biomedicine. Herein, we describe a lab-on-a-chip platform based on dielectrophoresis (DEP). The DEParray is a prototypal version consisting of 320 × 320 arrayed electrodes generating >10,000 spherical DEP cages. It allows the capture and software-guided movement to predetermined spatial coordinates of single biological objects. With the DEParray we demonstrate (a) forced interaction between a single, preselected target cell and a programmable number of either microspheres or natural killer (NK) cells, (b) on-chip immunophenotypic discrimination of individual cells based on differential rosetting with microspheres functionalized with monoclonal antibodies to an inhibitory NK cell ligand (HLA-G), (c) on-chip, real-time (few minutes) assessment of immune lysis by either visual inspection or semiautomated, time-lapse reading of a fluorescent dye released from NK cell-sensitive targets, and (d) manipulation and immunophenotyping with limiting amounts (about 500) cells. To our knowledge, this is the first report describing a DEP-based lab-on-a-chip platform for the quick, arrayed, software-guided binding of individually moved biological objects, the targeting of single cells with microspheres, and the real-time characterization of immunophenotypes. The DEParray candidates as a discovery tool for novel cell:cell interactions with no prior (immuno)phenotypic knowledge.

Model-based prediction of the alpha-hemolysin structure in the hexameric state.

The alpha-hemolysin toxin self-assembles in lipid bilayers to form water-filled pores. In recent years, alpha-hemolysin has received great attention, mainly due to its possible usage as a sensing element. We measured the ion currents through single alpha-hemolysin channels and confirmed the presence of two different subpopulations of channels with conductance levels of 465 +/- 30 pS and 280 +/- 30 pS. Different oligomerization states could be responsible for these two conductances. In fact, a heptameric structure of the channel was revealed by x-ray crystallography, whereas atomic force microscopy revealed a hexameric structure. Due to the low resolution of atomic force microscopy the atomic details of the hexameric structure are still unknown, and are here predicted by computational methods. Several possible structures of the hexameric channel were defined, and were simulated by molecular dynamics. The conductances of these channel models were computed by a numerical method based on the Poisson-Nernst-Planck electrodiffusion theory, and the values were compared to experimental data. In this way, we identified a model of the alpha-hemolysin hexameric state with conductance characteristics consistent with the experimental data. Since the oligomerization state of the channel may affect its behavior as a molecular sensor, knowing the atomic structure of the hexameric state will be useful for biotechnological applications of alpha-hemolysin.

Electronic sorting and recovery of single live cells from microlitre sized samples.

Sorting and recovering specific live cells from samples containing less than a few thousand cells have become major hurdles in rare cell exploration such as stem cell research, cell therapy and cell based diagnostics. We describe here a new technology based on a microelectronic chip integrating an array of over 100,000 independent electrodes and sensors which allow individual and parallel single cell manipulation of up to 10,000 cells while maintaining viability and proliferation capabilities. Manipulation is carried out using dynamic dielectrophoretic traps controlled by an electronic interface. We also demonstrate the capabilities of the chip by sorting and recovering individual live fluorescent cells from an unlabeled population.

Dielectrophoresis-based 'Lab-on-a-chip' devices for programmable binding of microspheres to target cells.

There is a general agreement on the fact that the Laboratory on chip (Lab-on-a-chip) technology will enable laboratory testing to move from laboratories employing complex equipments into non-laboratory settings. In this respect, dielectrophoresis (DEP) is a very valuable approach to design and produce Lab-on-a-chip devices able to manipulate microparticles and cells. In this study, we report the application of DEP-based devices for facilitating programmable interactions between microspheres and target tumor cells. We used two Lab-on-a-chip devices, one (the SmartSlide) carrying 193 parallel electrodes and generating up to 50 cylinder-shaped DEP cages, the other (the DEP array) carrying 102,400 arrayed electrodes and generating more than 10,000 spherical DEP cages. We determined whether these devices can be used to levitate and move microspheres and cells in order to obtain a forced interaction between microspheres and target cells. The first major point of this manuscript is that the DEP-based SmartSlide can be used for transfection experiments in which microspheres and target cells are forced to share the same DEP cage, leading to efficient binding of the microspheres to target cells. The data obtained using the DEP array show that this system allows the sequential, software-controlled binding of individually and independently moved single microspheres to a single target tumor cell. To our knowledge, this is the first report on the possible use of a DEP-based Lab-on-a-chip device for guided multiple binding of singularly moved microspheres to a single tumor cell. This approach can be of interest in the field of drug discovery, delivery and diagnosis.

Separation of white blood cells from erythrocytes on a dielectrophoresis (DEP) based 'Lab-on-a-chip' device.

The 'Lab-on-a-chip technology' involves miniaturization of complex analytical procedures and is expected to enable laboratory testing to move from the central laboratory employing complex equipment into non-laboratory settings. We report the application of a printed circuit board (PCB)-based chip, generating dielectrophoretic (DEP)-based cylinder-shaped cages for separation and recovery of white blood cells from erythrocytes. This possibility is of interest to develop low-cost Lab-on-a-chip devices for diagnostic purposes. Accordingly, we demonstrate that white blood cells recovered from this Lab-on-a-chip device are suitable for PCR-based molecular diagnosis procedures employing DNA sequencing or biospecific interaction analysis using surface plasmon resonance and biosensor technology.

Applications to cancer research of "lab-on-a-chip" devices based on dielectrophoresis (DEP).

The recent development of advanced analytical and bioseparation methodologies based on microarrays and biosensors is one of the strategic objectives of the so-called post-genomic. In this field, the development of microfabricated devices could bring new opportunities in several application fields, such as predictive oncology, diagnostics and anti-tumor drug research. The so called "Laboratory-on-a-chip technology", involving miniaturisation of analytical procedures, is expected to enable highly complex laboratory testing to move from the central laboratory into non-laboratory settings. The main advantages of Lab-on-a-chip devices are integration of multiple steps of different analytical procedures, large variety of applications, sub-microliter consumption of reagents and samples, and portability. One of the requirement for new generation Lab-on-a-chip devices is the possibility to be independent from additional preparative/analytical instruments. Ideally, Lab-on-a-chip devices should be able to perform with high efficiency and reproducibility both actuating and sensing procedures. In this review, we discuss applications of dielectrophoretic(DEP)-based Lab-on-a-chip devices to cancer research. The theory of dielectrophoresis as well as the description of several devices, based on spiral-shaped, parallel and arrayed electrodes are here presented. In addition, in this review we describe manipulation of cancer cells using advanced DEP-based Lab-on-a-chip devices in the absence of fluid flow and with the integration of both actuating and sensing procedures.