ARTICLE | doi:10.20944/preprints201901.0037.v1
Subject: Engineering, Electrical & Electronic Engineering Keywords: microwaves, microfluidic, sensor, LTCC
Online: 4 January 2019 (09:00:27 CET)
This paper compares two types of microfluidic sensors that are designed for operation in ISM bands at microwave frequencies of 2.45 GHz and 5.8 GHz. In the case of the first sensor, the principle of operation is based on the resonance phenomenon in a microwave circuit filled with a test sample. The second sensor is based on the interferometric principle and makes use of the superposition of two coherent microwave signals, where only one of them goes through a test sample. Both sensors are monolithic structures fabricated using low temperature co-fired ceramics (LTCC). The LTCC-based microwave-microfluidic sensor properties are examined and compared by measuring their responses for various concentrations of two types of test fluids: one is a mixture of water/ethanol, and the other is dopamine dissolved in a buffer solution. The experiments show a linear response for the LTCC-based microwave-microfluidic sensors as a function of the concentration of the components in both test fluids.
ARTICLE | doi:10.20944/preprints202103.0264.v2
Online: 12 March 2021 (11:29:04 CET)
Morphological dissimilarity and its evolution over time are one of the most unexpected variations found when comparing cell cultures in 2D and 3D. Monolayer cells appear to flatten in the lower part of the plate, adhering to and spreading in the horizontal plane while not extending vertically. Consequently, cells developed in two dimensions have a forced apex-basal polarity. Co-cultivation and crosstalking between multiple cell types, which control development and formation in the in vivo counterpart, are possible in 3D cultures. With or without a scaffold matrix, 3D model culture may exhibit more in vivo-like morphology and physiology. 3D cultures mimic relevant physiological cellular processes, transforming them into one-of-a-kind drug screening platforms. The structures and dynamics of regulatory networks, which are increasingly studied with live-imaging microscopy, must be considered to help and guarantee the functional maintenance of a 3D structure. However, commercially available technologies that can be used for current laboratory needs are minimal, despite the need to make it easier to acquire cellular kinetics with high spatial and temporal resolution, in order to improve visual efficiency and, as a result, experimentation performance. The CELLviewer is a newly developed multi-technology instrument that integrates and synchronizes the work of various scientific disciplines. The aim of this study is to test the device using two different models: a single Jurkat cell and an MCF-7 spheroid. The two models are loaded into the microfluidic cartridge for each experiment after they have been grown and captured in time-lapse for a total of 4 hours. The samples used are tracked under the operation of the optics after adaptive autofocus, while slipping inside the cartridge chamber, and the 3D rotation was successfully obtained experimentally. The MitoGreen dye, a fluorescence marker selectively permeable to live cells, was then used to determine cell viability. To measure the model diameter, construct fluorescence intensity graphs along a straight line passing through the cell, and visualize the spatial fluorescence intensity distribution in 3D, ImageJ software was used.
REVIEW | doi:10.20944/preprints202010.0622.v1
Subject: Engineering, Biomedical & Chemical Engineering Keywords: Cancerous Cell, Tumors, Cancer Metastasis, Circulating Tumor Cells (CTCs), Microfluidic Devices, lab-On-a-Chip (LOC), Lab-On-a-CD (LOCD)
Online: 29 October 2020 (15:38:54 CET)
Separation and interpretation of rebellious Circulating Tumor Cells (CTCs) originating from the primary tumor or cancer tissue plays a significant role in diagnostics, cancer progression analyses, suitable medicine exploration, and treatment proficiency examination. Cancer metastasis occurs when CTCs spread throughout the body and invade healthy tissues, leading to new tumors in that area. Although a dramatic rate of deaths begins from spreading CTCs around the body, valuable measures have been made to control their development. However, the first step is separating these harmful cells from the bloodstream and investigating their features. Having examined the characteristics of CTCs as cancer’s main strength, researchers can introduce complementary treatments that can affect cancerous cells without damaging the healthy cells. Therefore, according to their unique characteristics, numerous techniques have been established for continuous and fast separation and sorting of CTCs. Nevertheless, few separators enjoy the efficient performance and appropriate accuracy and can be produced in mass numbers due to the available fabrication equipment. Microfabrication advancements enable separators to combine the advantages of active and passive methods in a small-scale platform for probing individual cells and separation purposes. Reduction in reagents, sample volume, analysis time, and less harmfulness to patients are some of the motivations that encourage researchers to employ microfluidic instruments for CTCs separation from other blood cells over the last two decades. However, microfabrication limitations mean effective separators, and the diagnostic option they provide, are not readily available. Addressing these limitations requires optimizing the design and fabrication of separators such that they are reduced in their size and fabrication cost, while also maintaining high-throughput separating capability. The emergence of the Lab-On-a-Chip (LOC) and then Lab-On-a-CD (LOCD) technologies, having more inherent benefits than conventional microfluidic devices, has created new opportunities and become increasingly widespread in recent years. Evidence suggests that employing single methodologies or integrating approaches without sufficient understanding of potential outcomes is unlikely to result in successful diagnostic results. This paper contributes an extensive review of several separation systems, including fundamental theories and experimental details, and describes detailed operating principles and device performance.
Subject: Chemistry, Analytical Chemistry Keywords: Microfluidic, Conductivity, Mobil-phone, urea, urease enzyme
Online: 22 February 2021 (16:44:31 CET)
In this work, PMMA(Poly methyl methacrylate) microfluidic system was used as a micro-reactor for urea hydrolysis by urease enzyme with use conductivity principle and utilize sound level meter(SLM) App in smartphone as a novel detector by considering the peak height in the App as an indicate for urea concentration. the advantage of use small volume and how the reaction carried on in the microfluidic system with simple and low cost are discussed, and the results were analyzed and statically determine. The linearity, detection limit (3×noise )and Correlation Coefficient, 62.5-500 ppm, 31.25 ppm and 0.992 respectively also, recovery studied were between(98.5-100.13%).
ARTICLE | doi:10.20944/preprints201903.0031.v1
Subject: Keywords: Organ-on-Chip, tissue engineering, microfluidic technology
Online: 4 March 2019 (10:11:22 CET)
Organ-on-Chip is a game-changing technology born from the convergence of tissue engineering and microfluidic technology. Organ-on-Chip devices (OoCs) are expected to offer effective solutions to persisting problems in drug development and personalized disease treatments. This opinion paper surveys the current landscape in research, development, application and market opportunities for OoCs to help establishing a global and multi-stakeholder OoC ecosystem. Based on a bibliometric study, a market analysis, expert interviews, and panel discussions held at the ORCHID Vision Workshop (Stuttgart, 23 May 2018), we outline presently unmet needs, key challenges, barriers and perspectives of the field, and finally propose recommendations towards the definition of a comprehensive roadmap that could render OoCs realistic models of human (patho)physiology in the near future.
Subject: Physical Sciences, Acoustics Keywords: capillary; microfluidic device; single-molecule recycling; maximum likelihood
Online: 26 May 2021 (10:50:13 CEST)
Microfluidic devices have been extensively investigated in recent years in fields including ligand-binding analysis, chromatographic separation, molecular dynamics, and DNA sequencing. To prolong the observation of a single molecule in aqueous buffer, the solution in a sub-micron scale channel is driven by the electric field and reversed after a fixed delay following each passage, so that the molecule passes back and forth through the laser focus and the time before irreversible photobleaching is extended. However, this practice requires complex chemical treatment to the inner surface of the channel to prevent unexpected sticking to the surface and the confined space renders features, such as a higher viscosity and lower dielectric constant, which slow the Brownian motion of the molecule compared to the bulk liquid. In this paper, we have fixed a capillary microchannel with an inner diameter of 2 microns on top of a piezo stage to recycle the molecule and collected the fluorescence by a confocal microscope. The passing times of the molecule through the laser focus are calculated by a real-time control system based on an FPGA and the commands of translation are given to the piezo stage through a feedback algorithm. We have achieved a maximum number of recycles of more than 200 and developed a maximum-likelihood estimation of the diffusivity of the molecule, which attains results of the same magnitude as previous reports. This technique simplifies the overall procedure of the single-molecule recycling and could be useful for the ligand-binding studies of biomolecules.
ARTICLE | doi:10.20944/preprints201807.0232.v2
Subject: Engineering, Biomedical & Chemical Engineering Keywords: microfluidic; microelectrodes; negatively charged; TiO2; assisted reproductive technology
Online: 18 September 2018 (10:34:00 CEST)
This research was presented the special designed microfluidic device generated for sperm separation based on assumption of different surface electrical charged of sperms X and Y. However, to avoid ethical problem, the microfluidic chip has been tested with the mimic electrical charged particles, TiO2-coated Polystyrene beads, (TiO2-coated Ps-beads), instead of spermatozoa. The work has been separated into three main parts. Firstly, the simply but efficient fabrication of negatively charged TiO2-coated Ps-beads has been presented. In addition, various characterization techniques such as X-ray diffraction (XRD), Tungsten Scanning Electron Microscopy (W-SEM) with energy-dispersive X-ray spectroscopy (EDS) mode, and X-ray Absorption Spectroscopy (XAS), have been reported in this work to elucidate the reasons behind the persistence of negatively charged on the surface of TiO2-coated Ps-beads. Results show that the fabricated TiO2-coated Ps-beads was partly coated in the mixed forms of amorphous Ti4+ and had caused a negatively charge to appear on the surface after fabrication and had sustained its electrical charged for long. Secondly, process of simulation and fabrication of microfluidic device was presented. Finally the negatively charged TiO2-coated Ps-beads were tested in this microfluidic devices. For design of microfluidic devices integrated with a couple of microelectrodes, the simulated structures were fabricated by photolithographic technique and tested with the Ps-beads. Percentage of validation for Ps-beads separation indicated that the 100 mm-distance-between-electrodes microfluidic device exhibits to be the highest performance prototype at 86.96%. For further confirmation, another model so called the single path prototype has been established. It is confirmed by 92.59% of validation for the utilization of the device. The successfully designed microfluidic devices can be examined with actual spermatozoa later. Furthermore, process to fabricate the negatively charged TiO2-coated Ps-beads can be established as testified samples for development of other microfluidic devices.
ARTICLE | doi:10.20944/preprints202107.0597.v1
Subject: Engineering, Biomedical & Chemical Engineering Keywords: 3D printing; microfabrication; microfluidic guillotine; single cell; wound healing
Online: 27 July 2021 (09:21:04 CEST)
Micro-blade design is an important factor in the cutting of single cells and other biological structures. This paper describes the fabrication process of three dimensional (3D) micro-blades for the cutting of single cells in a microfluidic “guillotine” intended for fundamental wound repair and regeneration studies. Our microfluidic guillotine consists of a fixed 3D micro-blade centered in a microchannel to bisect cells flowing through. We show that the Nanoscribe two-photon polymerization direct laser writing system is capable of fabricating complex 3D micro-blade geometries. However, structures made of the Nanoscribe IP-S resin have low adhesion to silicon, and they tend to peel off from the substrate after at most two times of replica molding in poly(dimethylsiloxane) (PDMS). Our work demonstrates that the use of a secondary mold replicates Nanoscribe printed features faithfully for at least 10 iterations. Finally, we show that complex micro-blade features can generate different degrees of cell wounding and cell survival rates compared with simple blades possessing a vertical cutting edge fabricated with conventional 2.5D photolithography. Our work lays the foundation for future applications in single cell analyses, wound repair and regeneration studies, as well as investigations of the physics of cutting and the interaction between the micro-blade and biological structures.
ARTICLE | doi:10.20944/preprints202010.0506.v1
Subject: Chemistry, Analytical Chemistry Keywords: Paper-based microfluidic device; colorimetric; multiple detection; smartphone application
Online: 26 October 2020 (08:56:58 CET)
Paper-based microfluidic analysis devices (μPADs) have attracted attention as a cost-effective platform for point-of-care testing (POCT), food safety, and environmental monitoring. Recently, three-dimensional (3D)-μPADs have been developed to improve the performance of μPADs. For accurate diagnosis of diseases, however, 3D-μPADs need to be developed to simultaneously detect multiple biomarkers. Here, we report a 3D-μPADs platform for the detection of multiple biomarkers that can be analyzed and diagnosed with a smartphone. The 3D-μPADs were fabricated using a 3D digital light processing printer and consisted of a sample reservoir (300 µL) connected to 24 detection zones (of 4 mm in diameter) through 8 microchannels (of 2 mm in width). With the smartphone application, eight different biomarkers related to various diseases were detectable in concentrations ranging from normal to abnormal conditions: glucose (0–20 mmol/L), cholesterol (0–10 mmol/L), albumin (0–7 g/dL), alkaline phosphatase (0–800 U/L), creatinine (0–500 µmol/L), aspartate aminotransferase (0–800 U/L), alanine aminotransferase (0–1000 U/L), and urea nitrogen (0–7.2 mmol/L). These results suggest that 3D-µPADs can be used as a POCT platform for simultaneous detection of multiple biomarkers.
ARTICLE | doi:10.20944/preprints201811.0406.v1
Subject: Engineering, Biomedical & Chemical Engineering Keywords: SU-8, microchannel, prototyping, microfluidic gradient generator, axon elongation
Online: 16 November 2018 (11:19:18 CET)
We have developed a cast microfluidic chip for concentration gradient generation that contains a thin (~5 μm^2 crosssectional area) microchannel. Durable 2 μm-high microchannel mold features with a smooth bell-shaped sidewall were fabricated by exposing SU-8 photoresist to diffused 185 nm UV light emitted by a low-cost ozone lamp from the backside of the substrate to ensure sufficient crosslinking of small regions of the SU-8 photoresist. An H-shaped microfluidic configuration was used, in which the thin channel was able to maintain constant diffusion fronts beyond purely static diffusion confirmed with experiment. We also demonstrated the long-term effects of a gradient of nerve growth factor on axon elongation by primary neuronal cells cultured in the microfluidic channel.
ARTICLE | doi:10.20944/preprints201803.0268.v1
Subject: Engineering, Electrical & Electronic Engineering Keywords: surface modification; electroosmotic flow; microfluidic; silicon nanochannel; thermal oxidation
Online: 30 March 2018 (10:14:47 CEST)
A simple fabrication method in the surface modification of electroosmotic silicon microchannel using thermal dry oxidation is presented. The surface modification is done by coating the silicon surface with a silicon dioxide (SiO2) layer using thermal oxidation process. The process is aimed not only to improve the surface quality of the channel to be suitable for electroosmotic fluid transport but also to reduce the channel width using a simple technique. Initially, the parallel microchannel array with dimensions of 0.5 mm length and width ranging from 1.8 µm to 2 µm are created using plasma etching on the 2x2 cm <100> silicon substrate. The oxidation of silicon channel in a thermal chamber is then conducted to create the SiO2 layer. The layer properties and the quality of the surface are analyzed using SEM and surface profiler, respectively. The results show that the maximum oxidation growth rate occurs in the first 4 hours of oxidation time and the rate decreases by time as the oxide layer becomes thicker. It is also found that the surface roughness is reduced with the increase of process temperature and oxide thickness. The scallop effect on the vertical wall due to plasma etching process also improved with the presence of the oxide layer. After the oxidation, the channel width is reduced by ~40%. The demonstrated method is suggested for the fabrication of a uniform channel cross section with high aspect ratio in sub-micro and nanometer scale that will be useful for the electroosmotic flow (EOF) manipulation of the biomedical fluid sample.
ARTICLE | doi:10.20944/preprints201708.0028.v1
Subject: Physical Sciences, Condensed Matter Physics Keywords: crystal growth; calcite; microfluidic; nanoconfinement; reflection interference contrast microscopy.
Online: 8 August 2017 (08:09:11 CEST)
Slow growth of calcite in confinement is abundant in Nature and man made materials. There is ample evidence that such confined growth may create forces that fracture solids. The thermodynamic limits are well known but since confined crystal growth is transport limited and difficult to control in experiment we have almost no information on the mechanisms or limits of these processes. We present a novel approach to in situ study of confined crystal growth using microfluidics for accurate control of the saturation state of the fluid and interferometric measurement of the topography of the growing confined crystal surface. We observe and explain the diffusion limited confined growth structures observed and can measure the crystal "floating" on a fluid film of 10-40~nm thickness due to the disjoining pressure. We find that there are two end member behaviours: smooth or intermittent growth in the contact region, the latter being faster than the former.
ARTICLE | doi:10.20944/preprints202209.0372.v1
Subject: Physical Sciences, Optics Keywords: magnetic fluid; thermotropic nematic liquid crystals; terahertz modulation; microfluidic chip
Online: 23 September 2022 (10:27:05 CEST)
In recent years, solid state terahertz (THz) modulators have obtained rapid progress with the widespread use of two-dimensional (2D) materials in the field of THz; however, challenges remain in preparing flexible THz modulators. In this study, flexible ferromagnetic nematic materials were prepared by doping thermotropic nematic liquid crystals 5CB into magnetic fluids, and the influence of hydrogen bonding in water was reduced by a self-made cyclic olefin copolymer (COC) microfluidic chip. THz modulation characteristics of magnetic fluid and ferromagnetic nematic liquid crystal (FNLC) under the induction of external magnetic field were compared using a THz time domain spectroscopy system. Under the action of a 91 mT magnetic field, the magnetic fluid has a maximum modulation depth (MD) of 54%. Under the same magnetic field, the maximum MD of the ferromagnetic nematic liquid crystal materials increase to 78% because of the rearrangement of Fe3O4 nanoparticles induced by the topological defect of the liquid crystal. We demonstrate that the magneto-optical effect is significantly enhanced in the ferromagnetic nematic liquid crystal hybrid system. This strategy of doping thermotropic nematic liquid crystals to enhance the magneto-optical effect has great potential for THz filtering, modulation, and sensing applications.
ARTICLE | doi:10.20944/preprints201804.0363.v1
Subject: Physical Sciences, Optics Keywords: surface plasmon resonance; biosensing; nanofabrication; lab-on-a-chip; microfluidic
Online: 27 April 2018 (16:06:58 CEST)
Surface Plasmon Resonance (SPR) based sensors have the advantage of being label-free, enzyme-free and real-time. However, their spreading in multidisciplinary research is still limited and almost confined to prism-coupled devices. Plasmonic gratings, combined with a simple and cost-effective instrumentation, have been poorly developed compared to prism-coupled system mainly due to their lower sensitivity. Here we describe the optimization and signal enhancement of a sensing platform based on phase-interrogation method, which entails the exploitation of a nanostructured sensor. This technique is particularly suitable for integration of the plasmonic sensor in a lab-on-a-chip platform and can be used in a microfluidic circuit to ease the sensing procedures and limit the injected volume. The careful optimization of most suitable experimental parameters by numerical simulations leads to a 30 to 50% enhancement of SPR response, opening new possibilities for applications in the biomedical research field while maintaining the ease and versatility of the configuration.
ARTICLE | doi:10.20944/preprints202009.0259.v1
Subject: Engineering, Biomedical & Chemical Engineering Keywords: droplet breakup; microfluidic droplet generation; live image analysis; orifice; high pressure homogenization
Online: 11 September 2020 (12:46:49 CEST)
For our research on droplet deformation and breakup in scaled high-pressure homogenizing units we developed a pressure stable inline droplet generator. It consists of an optically accessible flow channel with a combination of stainless steel and glass capillaries and a 3D printed orifice. The droplet size is determined online by live image analysis. The influence of the orifice diameter, the mass flow of the continuous phase and the mass flow of the disperse phase on the droplet diameter was investigated. Furthermore, the droplet detachment mechanisms were identified. Droplet diameters with small diameter fluctuation between 175 µm and 500 µm could be realized, which allows a precise adjustment of the Ca and We Number in the subsequent scaled high pressure homogenizer disruption unit. The determined influence of geometry and process parameters on the resulting droplet size and droplet detachment mechanism agreed well with literature on microfluidics. Furthermore, droplet trajectories in an exemplary scaled high-pressure homogenizer disruption unit are presented which show that the droplets can be reinjected on a trajectory close to the center axis or close to the wall, which should result in different stresses on the droplets.
ARTICLE | doi:10.20944/preprints202109.0056.v1
Subject: Engineering, Biomedical & Chemical Engineering Keywords: Aptamer; Fluorescence Resonance Energy Transfer; Heavy Metal Ions; Graphene Oxide; Microfluidic Device; Sensor.
Online: 3 September 2021 (10:29:50 CEST)
Heavy metal contaminants have serious consequences for the environment and human health. Consequently, effective methods for detecting their presence, particularly in water and food, are urgently required. Accordingly, the present study proposes a sensor for the detection of mercury Hg(II) and lead Pb(II) ions using graphene oxide (GO) as a quenching agent and aptamer solu-tion as a reagent. In the proposed device, the aptamer sequences are labeled by FAM and HEX fluorescent dyes, respectively, and are mixed with 500 ppm GO solution in a microfluidic device. The presence of Hg(II) and Pb(II) ions is then detected by measuring the change in the fluores-cence intensity of the GO/aptamer suspension as the aptamer molecules undergo fluorescence resonance energy transfer (FRET). The experimental results show that the aptamer sensors have a linear range of 10~250 nM (i.e., 2.0~50 ppb) for Hg(II) ions and 10~100 nM (i.e., 2.1~20.7 ppb) for Pb(II) ions. Furthermore, the limit of detection is around 2 ppb for both metals, which is signifi-cantly lower than the maximum limits of 6 ppb and 10 ppb prescribed by the World Health Or-ganization (WHO) for Hg(II) and Pb(II) in drinking water, respectively.
ARTICLE | doi:10.20944/preprints201807.0605.v1
Subject: Chemistry, Electrochemistry Keywords: arrayed flexible chloride sensor; wireless sensing system; hysteresis voltage; selectivity coefficient; dynamic microfluidic
Online: 31 July 2018 (05:16:24 CEST)
Water quality monitoring was an important objective in the surroundings. In this study, we investigated the sensing characteristics of the arrayed flexible chloride sensor with XBee wireless sensing system. The sensitivity and linearity of the wireless chloride sensing devices were 91.6 mV/pCl and 0.988, respectively. The hysteresis voltages were 50.14 mV and 36.71 mV during the cycles of 1 M → 10−1 M → 1 M → 10−3 M → 1 M and 1 M → 10−3 M → 1 M → 10−1 M → 1 M, respectively. The selectivity coefficients of the ClO− ion, ClO4− ion, NO3− ion and I− ion for Cl− ion were 5.0 × 10−2, 1.0 × 10−1, 5.9 × 10−3 and 5.6×10−1, respectively. The sensing characteristics of real time measurement were investigated for dynamic microfluidic. The arrayed flexible chloride sensor was integrated with the microfluidic device, syringe pump and wireless sensing system. The sensitivity and linearity were 273.1 mV/pCl and 0.978 at 35 μL/min, respectively.
REVIEW | doi:10.20944/preprints201806.0011.v1
Subject: Engineering, Biomedical & Chemical Engineering Keywords: spheroid culture; microfluidic cell culture; spheroids on-chip; tumor microenvironment; in vitro cell culture
Online: 1 June 2018 (09:58:58 CEST)
Three-dimensional (3D) cell culture systems can be regarded as suitable platforms to bridge the huge gap between animal studies and two-dimensional (2D) monolayer cell culture to study chronic diseases such as cancer. In particular, the preclinical platforms for multicellular spheroid formation and culture can be regarded as ideal in vitro tumor models. The complex tumor microenvironment such as hypoxic region and necrotic core can be recapitulated in 3D spheroid configuration. Cells aggregated in spheroid structures can better illustrate the performance of anti-cancer drugs as well. Various methods have been proposed so far to create such 3D spheroid aggregations. Both conventional techniques and microfluidic methods can be used for generation of multicellular spheroids. In this review paper, we first discuss various spheroid formation phases. Then, the conventional spheroid formation techniques such as bioreactor flasks, liquid overlay and hanging droplet technique are explained. Next, a particular topic of the hydrogel in spheroid formation and culture is explored. This topic has received less attention in the literature. Hydrogels entail some advantages to the spheroid formation and culture such as size uniformity, the formation of porous spheroids or hetero-spheroids as well as chemosensitivity and invasion assays and protecting from shear stress. Finally, microfluidic methods for spheroid formation and culture are briefly reviewed.
ARTICLE | doi:10.20944/preprints201912.0172.v1
Subject: Life Sciences, Virology Keywords: ticks; cattle, rna viruses; next-generation sequencing; phylogeny; microfluidic real-time pcr technology; Caribbean; lips
Online: 12 December 2019 (12:29:36 CET)
Ticks transmit a wide variety of pathogens including bacteria, parasites and viruses. Over the last decade, numerous novel viruses have been described in arthropods, including ticks, and their characterization has provided new insights into RNA virus diversity and evolution. However, little is known about their ability to infect vertebrates. As very few studies have described the diversity of viruses present in ticks from the Caribbean, we implemented an RNA-sequencing approach on Amblyomma variegatum and Rhipicephalus microplus ticks collected from cattle in Guadeloupe and Martinique. Among the viral communities infecting Caribbean ticks, we selected four viruses belonging to the Chuviridae, Phenuiviridae and Flaviviridae families for further characterization and designing antibody screening tests. While viral prevalence in individual tick samples revealed high infection rates, suggesting a high level of exposure of Caribbean cattle to these viruses, no seropositive animals were detected. These results suggest that the Chuviridae- and Phenuiviridae-related viruses identified in the present study are more likely tick endosymbionts, raising the question of the epidemiological significance of their occurrence in ticks, especially regarding their possible impact on tick biology and vector capacity. The characterization of these viruses might open the door to new ways of preventing and controlling tick-borne diseases.
ARTICLE | doi:10.20944/preprints202109.0424.v1
Subject: Engineering, Biomedical & Chemical Engineering Keywords: high frequency dielectrophoresis; glioblastoma cells; single cell manipulation; microfluidic point-of-care device; cancer stem cells
Online: 24 September 2021 (11:11:35 CEST)
Glioblastoma (GBM) is one of the most aggressive solid tumors, particularly due to the presence of cancer stem cells (CSCs). Today the characterization of this type of cells with an efficient, fast and low-cost method remains an issue. Hence, we have developed a microfluidic lab-on-a-chip based on dielectrophoresis (DEP) single cell electro-manipulation to measure the two crossover frequencies: fx01 in low frequency range (below 500 kHz) and fx02 in Ultra High Frequency range (UHF, above 50 MHz). First, in vitro conditions were investigated. U87-MG cell lines were cultured in different conditions in order to induce an undifferentiated phenotype. Then, ex vivo GBM cells from patients’ primary cell culture, were passed through the developed microfluidic system and characterized in order to reflect clinical conditions. This article demonstrates that the usual exploitation of low frequency range DEP does not allow the discrimination of the undifferentiated from the differentiated phenotypes of GBM cells. However, the presented study highlights the use of UHF-DEP as a relevant discriminant parameter. The proposed microfluidic lab-on-a-chip is able to follow the kinetic of U87-MG phenotype transformation in a CSC enrichment medium and their cancer stem cells phenotype acquirement.
ARTICLE | doi:10.20944/preprints201611.0111.v1
Subject: Engineering, Biomedical & Chemical Engineering Keywords: Y-shaped microfluidic device; wall shear stress; adenosine triphosphate (ATP) signal; combined effect; vascular endothelial cells; calcium dynamics
Online: 22 November 2016 (09:51:31 CET)
The intracellular calcium dynamics in vascular endothelial cells (VECs) in response to wall shear stress (WSS) and/or adenosine triphosphate (ATP) have been commonly regarded as an important factor in regulating VEC function and behavior including proliferation, migration and apoptosis. However, the effects of time-varying ATP signals have been usually neglected in the past investigations in the field of VEC mechanobiology. In order to investigate the combined effects of WSS and dynamic ATP signals on the intracellular calcium dynamic in VECs, a Y-shaped microfluidic device, which can provide the cultured cells on the bottom of its mixing micro-channel with stimuli of WSS signal alone and different combinations of WSS and ATP signals in one single micro-channel, is proposed. Both numerical simulation and experimental studies verify the feasibility of its application. Cellular experimental results also suggest that a combination of WSS and ATP signals rather than a WSS signal alone might play a more significant role in VEC Ca2+ signal transduction induced by blood flow.
ARTICLE | doi:10.20944/preprints202007.0039.v1
Subject: Life Sciences, Biotechnology Keywords: Aviation security; biosensor; flow injection assay; monoclonal antibody; fluorescence microscope; lab-on-a-chip; microfluidic systems; antibody labeling; CMOS; diode laser; monolithic column; laser-induced fluorescence detector (LIF)
Online: 3 July 2020 (12:26:26 CEST)
The illegal use of explosives by terrorists and other criminals is an increasing issue in public spaces, such as airports, railway stations, highways, sports arenas, theaters, and other large buildings. Security in these environments can be achieved by a set of different means, including the installation of scanners and other analytical devices to detect ultra-small traces of explosives in a very short time-frame to be able to take action as early as possible to prevent the detonation of such devices. Unfortunately, an ideal explosive detection system still does not exist, which means that a compromise is needed in practice. Most detection devices lack the extreme analytical sensitivity, which is nevertheless necessary due to the low vapor pressure of nearly all explosives. In addition, the rate of false positives needs to be virtually zero, which is also very difficult to achieve. Here we present an immunosensor system based on kinetic competition, which is known to be very fast and may even overcome affinity limitation, which impairs the performance of many traditional competitive assays. This immunosensor consists of a monolithic glass column with a vast excess of immobilized hapten, which traps the fluorescently labeled antibody as long as no explosive is present. In the case of TNT occurring, some binding sites of the antibody will be blocked, which leads to an immediate breakthrough of the labeled protein, detectable by highly sensitive laser-induced fluorescence with the help of a Peltier-cooled CMOS camera. Liquid handling is performed with high-precision syringe pumps and chip-based mixing-devices and flow-cells. The system achieved limits of detection of 1 pM (1 ppt) of the fluorescent label and around 100 pM (20 ppt) of the explosive 2,4,6-trinitrotoluene (TNT). The total assay time is less than 8 min. A cross-reactivity test with 5000 pM solutions showed no signal by PETN, RDX, and HMX. This immunosensor belongs to the most sensitive and fastest detectors for TNT with no significant cross-reactivity by non-related compounds.
ARTICLE | doi:10.20944/preprints202107.0521.v1
Subject: Chemistry, Analytical Chemistry Keywords: Online detection, security; biosensor; flow injection assay; monoclonal antibody; fluorescence microscope; lab-on-a-chip; microfluidic systems; antibody labeling; CMOS; diode laser; monolithic column; laser-induced fluorescence detector (LIF); low-cost; high-speed; non-competitive immunoassay; immunometric assay
Online: 22 July 2021 (14:13:46 CEST)
The trafficking of illegal drugs by criminal networks at borders, harbors, or airports is an increasing issue in public health as these routes ensure the main supply of illegal drugs. The prevention of drug smuggling, including the installation of scanners and other analytical devices to detect ultra-small traces of drugs within a reasonable time frame, remains a challenge. The presented immunosensor is based on a monolithic affinity column with a large excess of immobilized hapten, which traps fluorescently labeled antibodies as long as the analyte cocaine is absent. In the presence of the drug, some binding sites of the antibody will be blocked, which leads to an immediate breakthrough of the labeled protein, detectable by highly sensitive laser-induced fluorescence with the help of a Peltier-cooled complementary metal-oxide-semiconductor (CMOS) camera. Liquid handling is performed with high-precision syringe pumps and microfluidic chip-based mixing devices and flow cells. The biosensor achieved limits of detection of 23 pM (7 ppt) of cocaine with a response time of 90 seconds and a total assay time below 3 minutes. With surface wipe sampling, the biosensor was able to detect 300 pg of cocaine. This immunosensor belongs to the most sensitive and fastest detectors for cocaine and offers near-continuous analyte measurement.