Characterization of an enzymatic packed-bed microreactor: Experiments and modeling

A micro packed-bed reactor (µPBR) based on two-parallel-plates configuration with immobilized Candida antarctica lipase B in the form of porous particles (Novozym® 435) was theoretically and experimentally characterized. A residence time distribution (RTD) within µPBRs comprising various random distributions of particles placed in one layer was computationally predicted by a mesoscopic lattice Boltzmann (LB) method. Numerical simulations were compared with measurements of RTD, obtained by stimulus-response experiment with a pulse input using glucose as a tracer, monitored by an electrochemical glucose oxidase microbiosensor integrated with the reactor. The model was validated by a good agreement between the experimental data and predictions of LB model at different conditions. The developed µPBR was scaled-up in length and width comprising either a single or two layers of Novozym® 435 particles and compared regarding the selected enzyme-catalyzed transesterification. A linear increase in the productivity with the increase in all dimensions of the µPBR between two-plates demonstrated very efficient and simple approach for the capacity rise. Further characterization of µPBRs of various sizes using the piezoresistive pressure sensor revealed very low pressure drops as compared to their conventional counterparts and thereby great applicability for production systems based on numbering-up approach

Lab-on-a-chip Thermoelectric and Solid-phase Immunodetection of Biochemical Analytes and Extracellular Vesicles: Experimental and Computational Analysis

Microfluidics is the technology of controlling and manipulating fluids at the microscale. Microfluidic platforms provide precise fluidic control coupled with low sample volume and an increase in the speed of biochemical reactions. Lab-on-a-chip platforms are used for detection and quantification of biochemical analytes, capture, and characterization of various proteins, sensitive analysis of cytokines, and isolation and detection of extracellular vesicles (EVs). This study focuses on the development of microfluidic and solid-phase capture pin platforms for the detection of cytokines, extracellular vesicles, and cell co-culture. The fabrication processes of the devices, experimental workflows, numerical analysis to identify optimal design parameters, and reproducibility studies have been discussed. Layer-by-layer assembly of polyelectrolytes has been developed to functionalize glass and stainless-steel substrates with biotin for the immobilization of streptavidinconjugated antibodies for selective capture of cytokines or EVs. Microstructure characterization techniques (SEM, EDX, and fluorescence microscopy) have been implemented to assess the efficiency of substrate functionalization. A detailed overview of current methods for purification and analysis of EVs is discussed as well. Additionally, the dissertation demonstrates the feasibility of a calorimetric microfluidic immunosensor with an integrated antimony-bismuth (Sb/Bi) thermopile sensor for the detection of cytokines with picomolar sensitivity. The developed platform can be used for the universal detection of both exothermic or endothermic reactions. A three-dimensional numerical model was developed to define the critical design parameters that enhance the sensitivity of the platform. Mathematical analyses identified the optimal combinations of substrate material and dimensions that will maximize the heat transfer to the sensor. Lab-on-a-chip cell co-culture platform with integrated pneumatic valve was designed, numerically characterized, and fabricated. This device enables the reversible separation of two cell culture chambers and serves as a tool for the effective analysis of cell-to-cell communication. Intercellular communication is mediated by extracellular vesicles. A protocol for the functionalization of stainless-steel probe with exosomespecific CD63 antibody was developed. The efficiency of the layer-by-layer deposition of polyelectrolytes and the effectiveness of biotin and streptavidin covalent boding were characterized using fluorescent and scanning electron microscopy

A nanostructured Fabry-Perot interferometer for label-free biodetection

A polymer nanostructured Fabry-Perot interferometer (FPI) based biosensor has been developed, fabricated, and tested. Different from a conventional FPI, this nanostructured FPI has a layer of Au-coated nanopores inside its cavity. The Au-coated nanostructure layer offers significant enhancement of optical transducing signals due to the localized surface Plasmon resonance (L-SPR) effect. Compared to a traditional FPI for label-free biosensing applications, the polymer nanostructured FPI based biosensor offers increased sensing surface area, extended penetration depth of the excitation light, and amplification of optical transducing signals. Using a nanostructured FPI, measurements taken had great improvements in free spectral range (FSR), finesse, and contrast of optical transducing signals over a traditional FPI without any device performance optimization. Several chemicals have been evaluated using the prototype device. Fourier Transform has been performed on the measured optical signals to facilitate the analysis of the transducing signals. Control experiments incubating immunoglobulin G (IgG) on a gold surface confirmed the small affinity of IgG to the Au-coated sensing surface. Then, using fluorescent images, shifts of interference fringes for IgG and BSA interaction were indirectly confirmed. Using this technical platform, the immobilization of capture proteins (Protein A) on the nanostructure layer and their binding with IgG was monitored in real time, resulting in the direct observation of the shift in interference fringes of the optical transducing signals. The results showed that the detection of limit (DOL) for this kind of biosensor should be lower than 10 pg/mL, which is approximately 55 fIVI of IgG, for IgG-Protein A binding. Control experiments were performed to confirm that the biodetection is only specific to Protein A and IgG recognition. After the proof-of-concept demonstration for IgG-Protein A binding, the ultrasensitive label-free detection of a cancer biomarker free prostate specific antigen (fPSA) using this kind of nanostructured FPI was carried out. Experiments found that the DOL of the fabricated nanostructured FPI microchip for f-PSA is about 5 pg/mL and the upper detection range for f-PSA can be dynamically changed by varying the amount of mAb immobilized on the sensing surface. Control experiments have also demonstrated that the immunoassay protocol used shows excellent specificity and selectivity, suggesting great potential to detect cancer biomarkers at trace levels in biofluids. Given its nature of low cost, simple operation, and batch fabrication capability, the nanostructured FPI microchip based platform could provide an ideal technical tool for point-of-care diagnostic applications and anti-cancer drug screening and discovery

Modelling microelectrode biosensors : free-flow calibration can substantially underestimate tissue concentrations

Microelectrode amperometric biosensors are widely used to measure concentrations of analytes in solution and tissue including acetylcholine, adenosine, glucose and glutamate. A great deal of experimental and modelling effort has been directed at quantifying the response of the biosensors themselves; however, the influence that the macroscopic tissue environment has on biosensor response has not been subjected to the same level of scrutiny. Here we identify an important issue in the way microelectrode biosensors are calibrated that is likely to have led to underestimations of analyte tissue concentrations. Concentration in tissue is typically determined by comparing the biosensor signal to that measured in free-flow calibration conditions. In a free-flow environment the concentration of the analyte at the outer surface of the biosensor can be considered constant. However, in tissue the analyte reaches the biosensor surface by diffusion through the extracellular space. Because the enzymes in the biosensor break down the analyte, a density gradient is set up resulting in a significantly lower concentration of analyte near the biosensor surface. This effect is compounded by the diminished volume fraction (porosity) and reduction in the diffusion coefficient due to obstructions (tortuosity) in tissue. We demonstrate this effect through modelling and experimentally verify our predictions in diffusive environments

Numerical Modeling of Amperometric Biosensor

Amperometric biosensor is a type of biosensor which measures the change in the current of a working indicator electrode by direct electrochemical oxidation or reduction of the products of a biochemical reaction. In these types of biosensors, the potential at the electrode is made constant during the measurement of current. These are known to be reliable, cheap and highly sensitive for environment, clinical and industrial purposes. These biosensors have plethora of applications in diverse fields; hence mathematical modeling of the same is highly desirable. This can help in prefiguring its various characteristics. A mathematical model is proposed which can study the cyclic conversion of substrate in an amperometric biosensor. The governing parameters for the Michaelis-Menten kinetics of enzymatic reactions are the enzyme kinetic rate and the diffusion rate across the enzymatic layer. Relative influence of these parameters is decided by a non dimensional number called Damkohler number, which is a ratio of the rate of enzymatic reaction to the rate of diffusion. The effect of Damkohler number on the current density, substrate concentration, and product concentration has been studied. It has been observed that when the Damkohler number is low then enzyme kinetics controls the biosensor response whereas when it is high (of the order of 1) the response is under control of diffusion rate. The current density is found to increase with the decrease in Damkohler number and vice versa

Microelectromechanical Systems and Devices

The advances of microelectromechanical systems (MEMS) and devices have been instrumental in the demonstration of new devices and applications, and even in the creation of new fields of research and development: bioMEMS, actuators, microfluidic devices, RF and optical MEMS. Experience indicates a need for MEMS book covering these materials as well as the most important process steps in bulk micro-machining and modeling. We are very pleased to present this book that contains 18 chapters, written by the experts in the field of MEMS. These chapters are groups into four broad sections of BioMEMS Devices, MEMS characterization and micromachining, RF and Optical MEMS, and MEMS based Actuators. The book starts with the emerging field of bioMEMS, including MEMS coil for retinal prostheses, DNA extraction by micro/bio-fluidics devices and acoustic biosensors. MEMS characterization, micromachining, macromodels, RF and Optical MEMS switches are discussed in next sections. The book concludes with the emphasis on MEMS based actuators

BioMEMS

As technological advancements widen the scope of applications for biomicroelectromechanical systems (BioMEMS or biomicrosystems), the field continues to have an impact on many aspects of life science operations and functionalities. Because BioMEMS research and development require the input of experts who use different technical languages and come from varying disciplines and backgrounds, scientists and students can avoid potential difficulties in communication and understanding only if they possess a skill set and understanding that enables them to work at the interface of engineering and biosciences. Keeping this duality in mind throughout, BioMEMS: Science and Engineering Perspectives supports and expedites the multidisciplinary learning involved in the development of biomicrosystems. Divided into nine chapters, it starts with a balanced introduction of biological, engineering, application, and commercialization aspects of the field. With a focus on molecules of biological interest, the book explores the building blocks of cells and viruses, as well as molecules that form the self-assembled monolayers (SAMs), linkers, and hydrogels used for making different surfaces biocompatible through functionalization. The book also discusses: Different materials and platforms used to develop biomicrosystems Various biological entities and pathogens (in ascending order of complexity) The multidisciplinary aspects of engineering bioactive surfaces Engineering perspectives, including methods of manufacturing bioactive surfaces and devices Microfluidics modeling and experimentation Device level implementation of BioMEMS concepts for different applications. Because BioMEMS is an application-driven field, the book also highlights the concepts of lab-on-a-chip (LOC) and micro total analysis system (μTAS), along with their pertinence to the emerging point-of-care (POC) and point-of-need (PON) applications

Modeling and Fundamental Design Considerations for Portable, Wearable and Implantable Electronic Biosensors

Chronic diseases such as cancer, diabetes, acquired immune deficiency syndrome (AIDS), etc. are leading causes of mortality all over the world. Portable, wearable and implantable biosensors can go a long way in preventing these premature deaths by frequent or continuous self-monitoring of vital health parameters

Micro/nanofluidic and lab-on-a-chip devices for biomedical applications

Micro/Nanofluidic and lab-on-a-chip devices have been increasingly used in biomedical research [1]. Because of their adaptability, feasibility, and cost-efficiency, these devices can revolutionize the future of preclinical technologies. Furthermore, they allow insights into the performance and toxic effects of responsive drug delivery nanocarriers to be obtained, which consequently allow the shortcomings of two/three-dimensional static cultures and animal testing to be overcome and help to reduce drug development costs and time [2–4]. With the constant advancements in biomedical technology, the development of enhanced microfluidic devices has accelerated, and numerous models have been reported. Given the multidisciplinary of this Special Issue (SI), papers on different subjects were published making a total of 14 contributions, 10 original research papers, and 4 review papers. The review paper of Ko et al. [1] provides a comprehensive overview of the significant advancements in engineered organ-on-a-chip research in a general way while in the review presented by Kanabekova and colleagues [2], a thorough analysis of microphysiological platforms used for modeling liver diseases can be found. To get a summary of the numerical models of microfluidic organ-on-a-chip devices developed in recent years, the review presented by Carvalho et al. [5] can be read. On the other hand, Maia et al. [6] report a systematic review of the diagnosis methods developed for COVID-19, providing an overview of the advancements made since the start of the pandemic. In the following, a brief summary of the research papers published in this SI will be presented, with organs-on-a-chip, microfluidic devices for detection, and device optimization having been identified as the main topics.info:eu-repo/semantics/publishedVersio

Spheroids-on-a-chip: Recent advances and design considerations in microfluidic platforms for spheroid formation and culture

© 2018 Elsevier B.V. A cell spheroid is a three-dimensional (3D) aggregation of cells. Synthetic, in-vitro spheroids provide similar metabolism, proliferation, and species concentration gradients to those found in-vivo. For instance, cancer cell spheroids have been demonstrated to mimic in-vivo tumor microenvironments, and are thus suitable for in-vitro drug screening. The first part of this paper discusses the latest microfluidic designs for spheroid formation and culture, comparing their strategies and efficacy. The most recent microfluidic techniques for spheroid formation utilize emulsion, microwells, U-shaped microstructures, or digital microfluidics. The engineering aspects underpinning spheroid formation in these microfluidic devices are therefore considered. In the second part of this paper, design considerations for microfluidic spheroid formation chips and microfluidic spheroid culture chips (μSFCs and μSCCs) are evaluated with regard to key parameters affecting spheroid formation, including shear stress, spheroid diameter, culture medium delivery and flow rate. This review is intended to benefit the microfluidics community by contributing to improved design and engineering of microfluidic chips capable of forming and/or culturing three-dimensional cell spheroids