Integration Of Microfluidics Research Articles

Lab-on-Chip Prototype Platform for Ochratoxin A Detection in Wine and Beer

We describe the development of a micro total analysis system for the detection of ochratoxin A (OTA) in wine and beer. The OTA detection strategy is based on an indirect competitive ELISA detection by chemiluminescence with the integration of microfluidics with microfabricated amorphous silicon photodiodes fabricated on glass substrates [1] . The full lab-on-chip (LoC) assay is implemented on a prototype device for fully-automatic measurement and monitoring of photodiode signals and control of fluid handling. The device is composed of three peripheral control and measurement modules: a peristaltic pump, a fluid dispenser for fluid handling, and a multi-sensor data acquisition with custom transimpedance amplifier. A microcontroller interfaces the individual modules with a computer via USB communication. The analysis is run automatically and the results displayed through a PC controlled user interface. OTA was detected in wine and beer extracts down to 0.5 ng/mL.

Optofluidic opportunities in global health, food, water and energy

Optofluidics is a rapidly advancing field that utilizes the integration of optics and microfluidics to provide a number of novel functionalities in microsystems. In this review, we discuss how this approach can potentially be applied to address some of the greatest challenges facing both the developing and developed world, including healthcare, food shortages, malnutrition, water purification, and energy. While medical diagnostics has received most of the attention to date, here we show that some other areas can also potentially benefit from optofluidic technology. Whenever possible we briefly describe how microsystems are currently used to address these problems and then explain why and how optofluidics can provide better solutions. The focus of the article is on the applications of optofluidic techniques in low-resource settings, but we also emphasize that some of these techniques, such as those related to food production, food safety assessment, nutrition monitoring, and energy production, could be very useful in well-developed areas as well.

Elastomer based tunable optofluidic devices

The synergetic integration of photonics and microfluidics has enabled a wide range of optofluidic devices that can be tuned based on various physical mechanisms. One such tuning mechanism can be realized based on the elasticity of polydimethylsiloxane (PDMS). The mechanical tuning of these optofluidic devices was achieved by modifying the geometry of the device upon applying internal or external forces. External or internal forces can deform the elastomeric components that in turn can alter the optical properties of the device or directly induce flow. In this review, we discuss recent progress in tunable optofluidic devices, where tunability is enabled by the elasticity of the construction material. Different subtypes of such tuning methods will be summarized, namely tuning based on bulk or membrane deformations, and pneumatic actuation.

Microfluidic very large scale integration (mVLSI) with integrated micromechanical valves

Microfluidic chips with a high density of control elements are required to improve device performance parameters, such as throughput, sensitivity and dynamic range. In order to realize robust and accessible high-density microfluidic chips, we have fabricated a monolithic PDMS valve architecture with three layers, replacing the commonly used two-layer design. The design is realized through multi-layer soft lithography techniques, making it low cost and easy to fabricate. By carefully determining the process conditions of PDMS, we have demonstrated that 8 × 8 and 6 × 6 μm(2) valve sizes can be operated at around 180 and 280 kPa differential pressure, respectively. We have shown that these valves can be fabricated at densities approaching 1 million valves per cm(2), substantially exceeding the current state of the art of microfluidic large-scale integration (mLSI) (thousands of valves per cm(2)). Because the density increase is greater than two orders of magnitude, we describe this technology as microfluidic very large scale integration (mVLSI), analogous to its electronic counterpart. We have captured and tracked fluorescent beads, and changed the electrical resistance of a fluidic channel by using these miniaturized valves in two different experiments, demonstrating that the valves are leakproof. We have also demonstrated that these valves can be addressed through multiplexing.

Lab-in-a-tube: on-chip integration of glass optofluidic ring resonators for label-free sensing applications

The fabrication of tubular rolled-up optofluidic ring resonators (RU-OFRRs) based on glass (SiO(2)) material with high quality factors is reported. A novel methodology combining lab-on-a-chip fabrication methods and rolled-up nanotech is presented for the fabrication of fully integrated tubular optofluidic sensors. The microfluidic integration of several RU-OFRRs on one chip is solved by enclosing the microtubes with a patterned robust SU-8 polymeric matrix. A viewport on each microtube enables exact excitation and monitoring of whispering gallery modes with a photoluminescence spectroscopy system under constant ambient conditions, while exchanging the content of the RU-OFRR with liquids of different refractive indices. The refractrometric sensor capabilities are investigated regarding signal stability, sensitivity and reliability. The sensitivity of the integrated RU-OFRR, which is the response of the modes to the change in refractive index of the liquid, is up to 880 nm/refractive index units (RIU).

Three-dimensional fit-to-flow microfluidic assembly

Three-dimensional microfluidics holds great promise for large-scale integration of versatile, digitalized, and multitasking fluidic manipulations for biological and clinical applications. Successful translation of microfluidic toolsets to these purposes faces persistent technical challenges, such as reliable system-level packaging, device assembly and alignment, and world-to-chip interface. In this paper, we extended our previously established fit-to-flow (F2F) world-to-chip interconnection scheme to a complete system-level assembly strategy that addresses the three-dimensional microfluidic integration on demand. The modular F2F assembly consists of an interfacial chip, pluggable alignment modules, and multiple monolithic layers of microfluidic channels, through which convoluted three-dimensional microfluidic networks can be easily assembled and readily sealed with the capability of reconfigurable fluid flow. The monolithic laser-micromachining process simplifies and standardizes the fabrication of single-layer pluggable polymeric modules, which can be mass-produced as the renowned Lego(®) building blocks. In addition, interlocking features are implemented between the plug-and-play microfluidic chips and the complementary alignment modules through the F2F assembly, resulting in facile and secure alignment with average misalignment of 45 μm. Importantly, the 3D multilayer microfluidic assembly has a comparable sealing performance as the conventional single-layer devices, providing an average leakage pressure of 38.47 kPa. The modular reconfigurability of the system-level reversible packaging concept has been demonstrated by re-routing microfluidic flows through interchangeable modular microchannel layers.

Optofluidic microsystems for chemical and biological analysis

Optofluidics - the synergistic integration of photonics and microfluidics - has recently emerged as a new analytical field that provides a number of unique characteristics for enhanced sensing performance and simplification of microsystems. In this review, we describe various optofluidic architectures developed in the past five years, emphasize the mechanisms by which optofluidics enhances bio/chemical analysis capabilities, including sensing and the precise control of biological micro/nanoparticles, and envision new research directions to which optofluidics leads.

Implantable microdevice for peripheral nerve regeneration: materials and fabrications

Rapid innovations in tissue engineering have increased the likelihood that fabricated microdevices for neuro-regeneration can finally be applied to humans. The advent of microdevices has created strong interest in many fields, including diagnostics, drug/gene delivery, and tissue engineering. The integration of microfluidics and tissue engineering is believed to hold promise in applications for neuro-regeneration. Early clinical results suggest that the fabrication of microdevices with appropriate properties indeed yields multi-functional applications with enhanced efficacy and less adverse effects. A prerequisite for advancing this area of research is the development of apt devices for nerve restoration, which can provide molecular, electrical, and micro-environmental cues for efficient neuronal cell regeneration. Based on the increasing clinical application of fabrication methods and continued efforts to advance this technology, it is likely that microdevice fabrication will become an effective method for achieving the above-mentioned criteria. Therefore, the aim of this review is to provide basic information on the fabrication of microdevices by focusing exclusively on several kinds of biomaterials, such as biocompatible, biodegradable, non-conducting, conducing, elastomeric and thermoplastic materials with natural, synthetic polymers, inorganic biomaterials, and physiochemical parameter for neuro-regeneration. We also discuss distinctive nerve growth factors and neural cell types within the context of developing micro-based neuro-degenerative applications. The information provided in this review is important with regards to the safe and widespread use of microdevice fabrication, particularly in the neuro-regenerative field.

Multianalyte On-Chip Native Western Blotting

We introduce and characterize multiplexed native Western blotting in an automated and unified microfluidic format. While slab gel Western blotting is slow and laborious, conventional multiplexed blotting ("reblotting": probing one sample with multiple antibodies) requires even more resources. Here we detail three key advances that enable an automated and rapid microfluidic alternative to slab gel reblotting. First, we introduce both assay and microdevice designs that integrate protein blotting against multiple antibody blotting regions with native polyacrylamide gel electrophoresis. This microfluidic integration strategy overcomes nonspecific material losses inherent to harsh antibody stripping steps typically needed for conventional reblotting; said conditions can severely limit analyte quantitation. Second, to inform rational design of the multiplexed microfluidic device we develop an analytical model for analyte capture on the blotting regions. Comparison to empirical observations is reported, with capture efficiencies of >85%. Third, we introduce label free detection that makes simultaneous and quantitative multiplexed measurements possible without the need for prelabeling of sample. Assay linear dynamic range spans 8-800 nM with assay completion in 5 min. Owing to the speed, automation, enhanced quantitation capability, and the difficulty of conventional slab gel Western reblotting, microfluidic multiplexed native Western blotting should find use in systems biology, in particular in analyses of protein isoforms and multimeric protein complexes.

A low-temperature parylene-to-silicon dioxide bonding technique for high-pressure microfluidics

We introduce a new low-temperature (280 °C) parylene-to-SiO2 bonding process with high device yield (>90%) for the fabrication and integration of high-pressure-rated microfluidic chips. Pull tests demonstrate a parylene-to-SiO2 bonding strength of 10 ± 3 MPa. We apply this technique for bonding Pyrex and silicon wafers having multiple metal layers to fabricate standard packaged microfluidic devices. By performing electrochemical impedance spectroscopy of electrolyte solutions in such devices, we demonstrate that electrodes remain functional after the etching, bonding and dicing steps. We also develop a high-pressure microfluidic and electrical integration technology, eliminating special fluidic interconnections and wire-bonding steps. The burst pressure of the integrated system is statistically shown to be 7.6 ± 1.3 MPa, with a maximum achieved burst pressure of 11.1 MPa, opening perspectives for high-pressure applications of these types of microfluidic devices.

Electrochemical Immunosensor for Simultaneous Detection of Dual Cardiac Markers Based on a Poly(Dimethylsiloxane)-Gold Nanoparticles Composite Microfluidic Chip: A Proof of Principle

The emergence of microfluidic immunosensors has provided a promising tool for improving clinical diagnoses. We developed an electrochemical immunoassay for the simultaneous detection of cardiac troponin I (cTnI) and C-reactive protein (CRP), based on microfluidic chips. The quantitative methodology was based on ELISA in poly(dimethylsiloxane)-gold nanoparticle composite microreactors. CdTe and ZnSe quantum dots were bioconjugated with antibodies for sandwich immunoassay. After the CdTe and ZnSe quantum dots were dissolved, Cd(2+) and Zn(2+) were detected by square-wave anodic stripping voltammetry to enable the quantification of the 2 biomarkers. The 2 biomarkers were measured in 20 human serum samples by using the proposed method and commercially available methods. This immunosensor allowed simultaneous detection of serum cTnI and CRP. The linear range of this assay was between 0.01 and 50 μg/L and 0.5 and 200 μg/L, with the detection limits of approximately 5 amol and approximately 307 amol in 30-μL samples corresponding to cTnI and CRP, respectively. Slopes close to 1 and the correlation coefficient over 0.99 were obtained for both analytes. This strategy demonstrates a proof of principle for the successful integration of microfluidics with electrochemistry that can potentially provide an alternative to protein detection in the clinical laboratory.

Microfluidic blood filtration device

Rapid decentralized biomedical diagnostics have become increasingly necessary in a medical environment of growing costs and mounting demands on healthcare personnel and infrastructure. Such diagnostics require low-cost novel devices that can operate at bedside or in doctor offices using small amounts of sample that can be extracted and processed on the spot. Thus, point-of-care sample preparation is an important component of the necessary diagnostic paradigm shift. We therefore introduce a microfluidic device which produces plasma from whole blood. The device is inexpensive, reliable, easy to fabricate, and requires only 3.5kPa pressure to operate. The device is fully compatible with microfluidic diagnostic chips. The output 23-gauge microtube of the former can be directly plugged into the input ports of the latter allowing immediate applicability in practice as a sample-prep pre-stage to a variety of emergent microfluidic diagnostic devices. In addition, the shown approach of filter encapsulation in elastomer has principle importance as it is compatible with and applicable to microfluidic sample-prep integration with analytical stages within the same elastomeric chip. This can eventually lead to finger-prick blood tests in point-of-care settings.

An automated instrument for human STR identification: Design, characterization, and experimental validation

The microfluidic integration of an entire DNA analysis workflow on a fully integrated miniaturized instrument is reported using lab-on-a-chip automation to perform DNA fingerprinting compatible with CODIS standard relevant to the forensic community. The instrument aims to improve the cost, duration, and ease of use to perform a "sample-to-profile" analysis with no need for human intervention. The present publication describes the operation of the three major components of the system: the electronic control components, the microfluidic cartridge and CE microchip, and the optical excitation/detection module. Experimental details are given to characterize the level of performance, stability, reliability, accuracy, and sensitivity of the prototype system. A typical temperature profile from a PCR amplification process and an electropherogram of a commercial size standard (GeneScan 500™, Applied Biosystems) separation are shown to assess the relevance of the instrument to forensic applications. Finally, we present a profile from an automated integrated run where lysed cells from a buccal swab were introduced in the system and no further human intervention was required to complete the analysis.

Microfluidic integration of nanomechanical resonators for protein analysis in serum

We describe a technique for reversibly encapsulating nanomechanical resonant sensors within microfluidic channels that are reusable and easy to fabricate. We demonstrate the feasibility and sensitivity of our microfluidic system by detecting prostate specific antigen (PSA) at 1–100 fM concentrations in serum, using a nanoparticle-based mass-labeling sandwich assay. By comparing these results to our previous data for PSA detection, we observe enhanced assay sensitivity in a significantly reduced time with the microfluidic assay. In addition, we show that a binary mixture of the proteins vascular endothelial growth factor (VEGF) and interleukin-8 (IL-8) can be detected in serum using a single microfluidic channel. This work lays the groundwork for future applications of these nanomechanical resonant devices for highly sensitive, multiplexed protein analysis using microfluidics.

Demonstration of the coupling of optofluidic ring resonator lasers with liquid waveguides

Optofluidic lasers are of particular interest for lab-on-a-chip-type devices, with broad spectral tunability, convenient microfluidic integration, and a small footprint. Optofluidic ring resonator (OFRR) lasers are advantageous in terms of size but typically generate nondirectional emission that is of minimal practical use. We introduce two unique geometries for soft-lithography-based OFRR lasers--side-coupled rings and spiral rings--both of which can be produced in polydimethyl siloxane substrates with contact molding. These rings utilize evanescent and direct butt-coupling, respectively, to effectively couple the OFRR laser emission into microfluidic channels. A laser threshold of a few to tens of microJ/mm(2) is achieved.

Single-step ELISA capillary sensor based on surface-bonded glucose oxidase, antibody, and physically-adsorbed PEG membrane containing peroxidase-labeled antibody

A single-step enzyme-linked immunosorbent assay (ELISA) capillary immunosensor that exploits the combination of surface-bonded glucose oxidase (GOD), antibody and physically-adsorbed poly(ethyleneglycol) (PEG) membrane containing peroxidase (POD)-labeled antibody in a single capillary, is presented. The present simple detection technique involves a proximity-based two-enzyme system where the product of the first enzyme reaction is used as a substrate into the second enzyme reaction. Since the degree of antigen–antibody reaction controls the second enzymatic reaction process, amount of final fluorescent product varies with antigen concentration that leads to the making ELISA procedure single step. Sample solution that contains antigen, glucose, ascorbic acid and a fluorescent substrate (amplex red) is simply introduced into the capillary immunosensor by capillary action. In the presence of an antigen in the nanoliter sample volume, the released POD-labeled antibody forms a sandwich immunocomplex on the surface bearing the antibody. This leads to a proximity of the POD and GOD. We confirmed that the rate of conversion of a non-fluorescent probe, amplex red, to a red fluorescent dye, resorufin, is dependent on the two-enzyme proximity in a certain experimental condition. Preliminary results yielded an approximate detection limit of around subnanogram per milliliter concentration for human IgG. The relative standard deviation of the fluorescence response was ca. 4.1% for the capillary pieces prepared by cutting a long immunosensor capillary. This approach could be an enabling technology applicable to microfluidic device integration for multiple analyte sensing. Realization of such an endeavor could be very promising for drug screening and clinical diagnostic applications.

Biosensors with label-free detection designed for diagnostic applications

Since the first biosensor was introduced in 1962 by Clark and Lyons, there has been increasing demand for such analytical devices in diagnostic applications. Research initially focussed mainly on detector principles and recognition elements, whereas the packaging of the biosensors and the microfluidic integration has been discussed only more recently. However, to obtain a user-friendly and well-performing analytical device, those components have to be considered all together. This review outlines the requirements and the solutions suggested for the integration of suitable biosensors in packaging and the integration of those encapsulated biosensors into a microfluidic surrounding resulting in a complete and efficient analytical device for diagnostic applications. The components required for a complete biosensor instrument are described and the latest developments which meet the requirements for diagnostic applications, such as single-use components and arrays for multiparameter detection, are discussed. The current state and the future of biosensors in the field of clinical diagnostics are outlined, particularly on the basis of label-free assay formats and the detection of prominent biomarkers for cancer and autoimmune disorders.

Three‐dimensional femtosecond laser micromachining of photosensitive glass for biomicrochips

AbstractInternal modification of transparent materials such as glass can be carried out using multiphoton absorption induced by a femtosecond (fs) laser. The fs‐laser modification followed by thermal treatment and successive chemical wet etching in a hydrofluoric (HF) acid solution forms three‐dimensional (3D) hollow microstructures embedded in photosensitive glass. This technique is a powerful method for directly fabricating 3D microfluidic structures inside a photosensitive glass microchip. We used fabricated microchips, referred to as a nanoaquarium, for dynamic observations of living microorganisms. In addition, the present technique can also be used to form microoptical components such as micromirrors and microlenses inside the photosensitive glass, since the fabricated structures have optically flat surfaces. The integration of microfluidics and microoptical components in a single glass chip yields biophotonic microchips, in other words, optofluidics, which provide high sensitivity in absorption and fluorescence measurements of small volumes of liquid samples.

Microfluidic lab-on-a-chip platforms: requirements, characteristics and applications

This critical review summarizes developments in microfluidic platforms that enable the miniaturization, integration, automation and parallelization of (bio-)chemical assays (see S. Haeberle and R. Zengerle, Lab Chip, 2007, 7, 1094-1110, for an earlier review). In contrast to isolated application-specific solutions, a microfluidic platform provides a set of fluidic unit operations, which are designed for easy combination within a well-defined fabrication technology. This allows the easy, fast, and cost-efficient implementation of different application-specific (bio-)chemical processes. In our review we focus on recent developments from the last decade (2000s). We start with a brief introduction into technical advances, major market segments and promising applications. We continue with a detailed characterization of different microfluidic platforms, comprising a short definition, the functional principle, microfluidic unit operations, application examples as well as strengths and limitations of every platform. The microfluidic platforms in focus are lateral flow tests, linear actuated devices, pressure driven laminar flow, microfluidic large scale integration, segmented flow microfluidics, centrifugal microfluidics, electrokinetics, electrowetting, surface acoustic waves, and dedicated systems for massively parallel analysis. This review concludes with the attempt to provide a selection scheme for microfluidic platforms which is based on their characteristics according to key requirements of different applications and market segments. Applied selection criteria comprise portability, costs of instrument and disposability, sample throughput, number of parameters per sample, reagent consumption, precision, diversity of microfluidic unit operations and the flexibility in programming different liquid handling protocols (295 references).

Fundamentals of microfluidic cell culture in controlled microenvironments

Microfluidics has the potential to revolutionize the way we approach cell biology research. The dimensions of microfluidic channels are well suited to the physical scale of biological cells, and the many advantages of microfluidics make it an attractive platform for new techniques in biology. One of the key benefits of microfluidics for basic biology is the ability to control parameters of the cell microenvironment at relevant length and time scales. Considerable progress has been made in the design and use of novel microfluidic devices for culturing cells and for subsequent treatment and analysis. With the recent pace of scientific discovery, it is becoming increasingly important to evaluate existing tools and techniques, and to synthesize fundamental concepts that would further improve the efficiency of biological research at the microscale. This tutorial review integrates fundamental principles from cell biology and local microenvironments with cell culture techniques and concepts in microfluidics. Culturing cells in microscale environments requires knowledge of multiple disciplines including physics, biochemistry, and engineering. We discuss basic concepts related to the physical and biochemical microenvironments of the cell, physicochemical properties of that microenvironment, cell culture techniques, and practical knowledge of microfluidic device design and operation. We also discuss the most recent advances in microfluidic cell culture and their implications on the future of the field. The goal is to guide new and interested researchers to the important areas and challenges facing the scientific community as we strive toward full integration of microfluidics with biology.