Various on-chip sensors with microfluidics for biological applications.

In this paper, we review recent advances in on-chip sensors integrated with microfluidics for biological applications. Since the 1990s, much research has concentrated on developing a sensing system using optical phenomena such as surface plasmon resonance (SPR) and surface-enhanced Raman scattering (SERS) to improve the sensitivity of the device. The sensing performance can be significantly enhanced with the use of microfluidic chips to provide effective liquid manipulation and greater flexibility. We describe an optical image sensor with a simpler platform for better performance over a larger field of view (FOV) and greater depth of field (DOF). As a new trend, we review consumer electronics such as smart phones, tablets, Google glasses, etc. which are being incorporated in point-of-care (POC) testing systems. In addition, we discuss in detail the current optical sensing system integrated with a microfluidic chip.

Pysanky to Microfluidics: An Innovative Wax-Based Approach to Low Cost, Rapid Prototyping of Microfluidic Devices.

A wax-based contact printing method to create microfluidic devices is demonstrated. This printing technology demonstrates a new pathway to rapid, cost-effective device prototyping, eliminating the use of expensive micromachining equipment and chemicals. Derived from the traditional Ukrainian Easter egg painting technique called "pysanky" a series of microfluidic devices were created. Pysanky is the use of a heated wax stylus, known as a "kistka", to create micro-sized, intricate designs on the surface of an egg. The proposed technique involves the modification of an x-y-z actuation translation system with a wax extruder tip in junction with Polydimethysiloxane (PDMS) device fabrication techniques. Initial system optimization was performed considering design parameters such as extruder tip size, contact angle, write speed, substrate temperature, and wax temperature. Channels created ranged from 160 to 900 µm wide and 10 to 150 µm high based upon system operating parameters set by the user. To prove the capabilities of this technology, a series of microfluidic mixers were created via the wax technique as well as through traditional photolithography: a spiral mixer, a rainbow mixer, and a linear serial dilutor. A thermo-fluidic computational fluid dynamic (CFD) model was generated as a means of enabling rational tuning, critical to the optimization of systems in both normal and extreme conditions. A comparison between the computational and experimental models yielded a wax height of 57.98 µm and 57.30 µm, respectively, and cross-sectional areas of 11,568 µm2 and 12,951 µm2, respectively, resulting in an error of 1.18% between the heights and 10.76% between the cross-sectional areas. The device's performance was then compared using both qualitative and quantitative measures, considering factors such as device performance, channel uniformity, repeatability, and resolution.

Review of imaging test phantoms.

Significance: Photoacoustic tomography has emerged as a prominent medical imaging technique that leverages its hybrid nature to provide deep penetration, high resolution, and exceptional optical contrast with notable applications in early cancer detection, functional brain imaging, drug delivery monitoring, and guiding interventional procedures. Test phantoms are pivotal in accelerating technology development and commercialization, specifically in photoacoustic (PA) imaging, and can be optimized to achieve significant advancements in PA imaging capabilities. Aim: The analysis of material properties, structural characteristics, and manufacturing methodologies of test phantoms from existing imaging technologies provides valuable insights into their applicability to PA imaging. This investigation enables a deeper understanding of how phantoms can be effectively employed in the context of PA imaging. Approach: Three primary categories of test phantoms (simple, intermediate, and advanced) have been developed to differentiate complexity and manufacturing requirements. In addition, four sub-categories (tube/channel, block, test target, and naturally occurring phantoms) have been identified to encompass the structural variations within these categories, resulting in a comprehensive classification system for test phantoms. Results: Based on a thorough examination of literature and studies on phantoms in various imaging modalities, proposals have been put forth for the development of multiple PA-capable phantoms, encompassing considerations related to the material composition, structural design, and specific applications within each sub-category. Conclusions: The advancement of novel and sophisticated test phantoms within each sub-category is poised to foster substantial progress in both the commercialization and development of PA imaging. Moreover, the continued refinement of test phantoms will enable the exploration of new applications and use cases for PA imaging.

An Integrated Centrifugal Degassed PDMS-Based Microfluidic Device for Serial Dilution.

We propose an integrated serial dilution generator utilizing centrifugal force with a degassed polydimethylsiloxane (PDMS) microfluidic device. Using gas-soluble PDMS as a centrifugal microfluidic device material, the sample can be dragged in any arbitrary direction using vacuum-driven force, as opposed to in a single direction, without adding further actuation components. The vacuum-driven force allows the device to avoid the formation of air bubbles and exhibit high tolerance in the surface condition. The device was then used for sample metering and sample transferring. In addition, centrifugal force was used for sample loading and sample mixing. In this study, a series of ten-fold serial dilutions ranging from 100 to 10-4 with about 8 µL in each chamber was achieved, while the serial dilution ratio and chamber volume could easily be altered by changing the geometrical designs of the device. As a proof of concept of our hybrid approach with the centrifugal and vacuum-driven forces, ten-fold serial dilutions of a cDNA (complementary DNA) sample were prepared using the device. Then, the diluted samples were collected by fine needles and subject to a quantitative polymerase chain reaction (qPCR), and the results were found to be in good agreement with those for samples prepared by manual pipetting.

2-layer based microfluidic concentration generator by hybrid serial and volumetric dilutions.

We present a 2-layer based microfluidic concentration generator by a hybrid of a serial and a volumetric dilution for dose-response experiments in drug screening. The hybrid dilution method using 2-layer based microfluidic network significantly reduces the total number of cascaded serial dilution stages. The proposed strategy is capable of generating a large number of universal stepwise monotonic concentrations with a wide range of logarithmic and linear scales. We have studied an equivalent electrical circuit to that of the 2-layer based microfluidic network, where the only variable parameter is channel length. We have designed a microfluidic dilution generator simultaneously covering 14 doses with a combination of 4-order logarithmic and 4-point linear concentrations. The design has been verified by a commercial circuit analysis software (e.g., P-Spice) for the electrical circuit analysis and a computational fluid dynamics software (e.g., CFD-ACE+) for the microfluidic circuit analysis. As a real-life application of the proposed dilution generator, we have successfully performed a dose-response experiment using MCF-7 human breast cancer cells. We expect that the proposed dilution method will be useful to study not only high throughput drug screening but also optimization in biology, chemistry, medicine, and material sciences.

Microfluidic Devices for Biomedical Applications: Biomedical Microfluidic Devices 2019.

Microfluidic devices and systems are well-suited for the manipulation of biomolecules, cells, or particles [...].

Passive micropumping in microfluidics for point-of-care testing.

Suitable micropumping methods for flow control represent a major technical hurdle in the development of microfluidic systems for point-of-care testing (POCT). Passive micropumping for point-of-care microfluidic systems provides a promising solution to such challenges, in particular, passive micropumping based on capillary force and air transfer based on the air solubility and air permeability of specific materials. There have been numerous developments and applications of micropumping techniques that are relevant to the use in POCT. Compared with active pumping methods such as syringe pumps or pressure pumps, where the flow rate can be well-tuned independent of the design of the microfluidic devices or the property of the liquids, most passive micropumping methods still suffer flow-control problems. For example, the flow rate may be set once the device has been made, and the properties of liquids may affect the flow rate. However, the advantages of passive micropumping, which include simplicity, ease of use, and low cost, make it the best choice for POCT. Here, we present a systematic review of different types of passive micropumping that are suitable for POCT, alongside existing applications based on passive micropumping. Future trends in passive micropumping are also discussed.

Dual Scan Mammoscope (DSM)-A New Portable Photoacoustic Breast Imaging System With Scanning in Craniocaudal Plane.

OBJECTIVE: We present a new photoacoustic tomography system that provides visualization of angiographic features in a human breast with mammogram-like images. METHODS: The system images a mildly compressed breast, from both top and bottom, using two 128-element, 2.25 MHz linear transducer arrays and line optical fiber bundles. The mild compression is achieved using plastic films, which is a more comfortable experience for the patient compared to rigid metal plates used in a traditional mammogram. RESULTS: We could image a D cup-sized breast of 7 cm thickness within 1 minute and achieve a spatial resolution of around 1 mm in all directions. CONCLUSION: Our system possesses the benefits of portability, speedy scanning, and patient comfort. The craniocaudal-view images can be easily correlated with existing imaging modalities for data interpretation. SIGNIFICANCE: Early cancer detection plays a critical role in overall cancer survival rate. Our system may address the limitations of mammogram and offer a radiation-free screening technique for patients with dense breasts.

Generalized serial dilution module for monotonic and arbitrary microfluidic gradient generators.

In this paper, we propose a generalized serial dilution module for universal microfluidic concentration gradient generators including N cascaded-mixing stages in a stepwise manner. Desired concentrations were generated by means of controlled volumetric mixing ratios of two merging solutions in each stage. The flow rates were adjusted by controlling channel length, which is proportional to fluidic resistance in each channel. A generalized mathematical model for generating any complex concentration and output flow rate gradients is presented based on the fact that there is an analogy between microfluidic circuits and electrical circuits. The pressure drop corresponds to a voltage drop, the flow rate to an electrical current, and the flow resistance to an electrical resistance. A simple equivalent electrical circuit model was generalized, and in the model each channel segment was represented by an electrical resistance. As a result of the mathematical modelling, the only variable parameter in the generalized serial dilution module was the channel length. By the use of the generalized serial dilution module with N = 4, three types of microfluidic gradient generators for linear, logarithmic and Gaussian gradients were successfully designed and tested. The proposed strategy is capable of generating universal monotonic gradients with a single module or arbitrary gradients with multiple modules ranging from linear to complex non-linear shapes of concentration gradients as well as arbitrary output flow rate gradients in a stepwise manner. The simple universal gradient generation technology using the generalized serial dilution module will find widespread use in the greater chemical and biological community, and address many challenges of gradient-dependent phenomena.

A Compact, Syringe-Assisted, Vacuum-Driven Micropumping Device.

In this paper, a simple syringe­assisted pumping method is introduced. The proposed fluidic micropumping system can be used instead of a conventional pumping system which tends to be large, bulky, and expensive. The micropump was designed separately from the microfluidic channels and directly bonded to the outlet of the microfluidic device. The pump components were composed of a dead­end channel which was surrounded by a microchamber. A syringe was then connected to the pump structure by a short tube, and the syringe plunger was manually pulled out to generate low pressure inside the microchamber. Once the sample was loaded in the inlet, air inside the channel diffused into the microchamber through the PDMS (polydimethylsiloxane) wall, acting as a dragging force and pulling the sample toward the outlet. A constant flow with a rate that ranged from 0.8 nl · s - 1 to 7.5 nl · s - 1 was achieved as a function of the geometry of the pump, i.e., the PDMS wall thickness and the diffusion area. As a proof-of-concept, microfluidic mixing was demonstrated without backflow. This method enables pumping for point-of-care testing (POCT) with greater flexibility in hand-held PDMS microfluidic devices.

Single-Layered Microfluidic Network-Based Combinatorial Dilution for Standard Simplex Lattice Design.

In this paper, we presented a straightforward strategy to generate 15 combinations of three samples based on an experimental simplex lattice design using a single-layer microfluidic network. First, we investigated the performances of the plain structural and the groove structural combinatorial devices by computational simulation (CFD-ACE+). The simulated output concentrations were extremely close to the desirable values within an absolute error of less than 1%. Based on the simulated designs, polydimethylsiloxane (PDMS) devices were fabricated with soft lithography and tested with fluorescent dye (sodium salt). The mixing results for 15 combinations showed good performance, with an absolute error of less than 4%. We also investigated two liquid handling methods (bottom⁻up and top⁻down) for high-throughput screening and assay. The liquid-handling methods were successfully accomplished by adding the systematic structured groove sets on the mixing channels.

A robust, portable and backflow-free micromixing device based on both capillary- and vacuum-driven flows.

In capillary- or vacuum-driven microfluidics, surge backflow events are common when merging or pumping two similar or dissimilar liquids together if a pressure difference exists between them. In this work, a robust, portable micromixing device that is insensitive to backflow was designed, fabricated and characterised. A capillary-driven pressure balancing bypass connected between two inlet ports diminished the initial pressure difference caused by capillarity and gravity present in each liquid at the two inlet ports. Then, using manual syringe-assisted vacuum-driven pumping that operated based on the high gas permeability of polydimethylsiloxane, the two pre-balanced liquid streams could synchronously enter a dead-end micromixing channel without any backflow. To test the performance of this device, we first used it to mix two aqueous solutions of different coloured dyes. We varied the initial volume difference between the solutions to study the effect of gravity-induced pressure difference on mixing. Next, as a proof-of-concept application, ABO/Rh blood groups were successfully determined through detection of blood antigen-antibody agglutination. The filling time of agglutinated samples, driven by the simple syringe-assisted pumping, in the dead-end mixing channel was consistently 10% longer than that of blood samples without the agglutination reaction. Thus, the proposed device shows great potential for use in a wide variety of blood typing assays, agglutination-based assays and point-of-care or lab-on-a-chip testing applications.

Manipulation of micro-objects using acoustically oscillating bubbles based on the gas permeability of PDMS.

This paper presents a novel manipulation method for micro-objects using acoustically oscillating bubbles with a controllable position based on the gas permeability of polydimethylsiloxane. The oscillating bubble trapped within the side channel attracts the neighboring micro-objects, and the position of the air-liquid interface is controlled by generating temporary pressure difference between the side channel and the air channel. To demonstrate the feasibility of the method in technological applications, polystyrene microparticles of 10 µm in diameter were successfully captured, transported, and released. The influence of pressure difference on the movement speed of the air-liquid interface was demonstrated in our experiments, and the manipulation performance was also characterized by varying the frequency of the acoustic excitation and the pressure difference. Since the bubble generation and the air-liquid interface movement in our manipulation method do not need any electrochemical reaction and any high temperature, this on-chip manipulation method provides a controllable, efficient, and noninvasive tool for handling micro-objects such as particles, cells, and other entities. The whole manipulation process, including capturing, transporting, and releasing of particles, spent less than 1 min. It can be used to select the cells and particles in the microfluidic device or change the cell culture medium.

Optimizing the light delivery of linear-array-based photoacoustic systems by double acoustic reflectors.

Although linear transducer arrays have been intensely used in photoacoustic imaging, their geometrical shape constrains light illumination. Today, most linear array based photoacoustic systems utilize side-illumination geometry, which consists of two line fiber bundles attached to the side of the probe. The angled light illumination increases the light travel distance in deep tissue, consequently limiting the imaging depth. This issue was partially addressed by adding a right angle prism in front of the transducer. While this design makes the light illumination and acoustic detection co-axial, the transducer and the fiber bundles are orthogonal to each other, making the system inconvenient for handheld use. To overcome this limitation, here we propose a double-reflector design, in which the second reflector redirects the acoustic signals by another 90°, so that the transducer and the fiber bundle are now parallel to each other. In this design, both the transducer and fiber bundle output are fitted into a compact housing for convenient handheld imaging. To evaluate the efficiency of our design, we performed various phantom and human in vivo experiments. Our results demonstrate that the double-reflector design indeed provides deeper imaging depth and it also allows for easy imaging of objects with uneven surfaces.

Multidisciplinary Role of Microfluidics for Biomedical and Diagnostic Applications: Biomedical Microfluidic Devices.

Life scientists are closely working with engineers to solve biological and biomedical problems through the application of engineering tools.[...].

Introduction of a Chemical-Free Metal PDMS Thermal Bonding for Fabrication of Flexible Electrode by Metal Transfer onto PDMS.

Polydimethylsiloxane (PDMS) is a flexible and biocompatible material widely used in the fabrication of microfluidic devices, and is often studied for the fabrication of flexible electrodes. The most popular method of fabricating a flexible electrode using PDMS is done by transferring a metal electrode onto said PDMS. However, the transfer process is difficult and the transferred metal layer is easily damaged due to inherently weak adhesion forces between the metal and PDMS, thus requiring a chemical treatment or sacrificial layer between the two. The fabrication process using a chemical treatment or sacrificial layer is complicated and expensive, which is the major limitation of using PDMS in the fabrication of flexible electrodes. This paper discusses the findings of a possible solution to create strong bonding between PDMS and various metals (copper, nickel and silver) using a chemical-free metal to PDMS thermal bonding technique. This method is the same as the PDMS curing process, but with a variation in the curing condition. The condition required to create strong bonding was studied by observing copper transferred by various PDMS curing conditions, including the standard condition. The condition creating the strong bonding was baking PDMS (5:1 = base polymer: curing agent) at 150 °C for 20 min. Experimentation showed that the optimum thickness of the transferred metal shows that the optimum thickness is approximately 500 nm, which allows for a higher resistance to stresses. The successful transfer of copper, nickel and silver layers onto PDMS with a stronger adhesion force opens up many new applications dealing with the fabrication of flexible electrodes, sensors, and flexible soft magnets.

Clinical evaluation of micro-scale chip-based PCR system for rapid detection of hepatitis B virus.

The polymerase chain reaction (PCR) is widely used to amplify a small amount of DNA in samples for genetic analysis. Rapid and accurate amplification is prerequisite for broad applications including molecular diagnostics of diseases, food safety, and biological warfare tests. We have developed a rapid real-time micro-scale chip-based PCR system, which consists of six individual thermal cycling modules capable of independent control of PCR protocols. The PCR volume is 1 microl and it takes less than 20 min to complete 40 thermal cycles. To test utility of a chip-based PCR system as a molecular diagnostic device, we have conducted the first large-scale clinical evaluation study. Three independent clinical evaluation studies (n = 563) for screening the hepatitis B virus (HBV) infection, the most popular social epidemic disease in Asia, showed an excellent sensitivity, e.g. 94%, and specificity, e.g. 93%, demonstrating micro-scale chip-based PCR can be applied in molecular diagnostics.

A Simple Method for Fabrication of Microstructures Using a PDMS Stamp.

We report a simple method to fabricate PDMS (polydimethylsiloxane) microwell arrays on glass by using a PDMS stamp to study cell-to-cell adhesion. In the cell-to-cell study, a glass substrate is required since glass has better cell attachment. The microwell arrays are replicated from an SU-8 master mold, and then are transferred to a glass substrate by lifting the PDMS stamp, followed by oxygen plasma bonding of the PDMS stamp on the glass substrate. For the cell-to-cell adhesion, four different types of PDMS arrays (e.g., rectangle, bowtie, wide-rhombus, and rhombus) were designed to vary the cell-to-cell contact length. The transfer success rates of the microwell arrays were measured as a function of both the contact area of the PDMS and the glass substrate and the different ratios between the base polymers and the curing agent. This method of generating the microwell arrays will enable a simple and robust construction of PDMS-based devices for various biological applications.

World-to-chip microfluidic interface with built-in valves for multichamber chip-based PCR assays.

We report a practical world-to-chip microfluidic interfacing method with built-in valves suitable for microscale multichamber chip-based assays. One of the primary challenges associated with the successful commercialization of fully integrated microfluidic systems has been the lack of reliable world-to-chip microfluidic interconnections. After sample loading and sealing, leakage tests were conducted at 100 degrees C for 30 min and no detectable leakage flows were found during the test for 100 microchambers. To demonstrate the utility of our world-to-chip microfluidic interface, we designed a microscale PCR chip with four chambers and performed PCR assays. The PCR results yielded a 100% success rate with no contamination or leakage failures. In conclusion, we have introduced a simple and inexpensive microfluidic interfacing system for both sample loading and sealing with no dead volume, no leakage flow and biochemical compatibility.

Vacuum-driven power-free microfluidics utilizing the gas solubility or permeability of polydimethylsiloxane (PDMS).

Suitable pumping methods for flow control remain a major technical hurdle in the path of biomedical microfluidic systems for point-of-care (POC) diagnostics. A vacuum-driven power-free micropumping method provides a promising solution to such a challenge. In this review, we focus on vacuum-driven power-free microfluidics based on the gas solubility or permeability of polydimethylsiloxane (PDMS); degassed PDMS can restore air inside itself due to its high gas solubility or gas permeable nature. PDMS allows the transfer of air into a vacuum through it due to its high gas permeability. Therefore, it is possible to store or transfer air into or through the gas soluble or permeable PDMS in order to withdraw liquids into the embedded dead-end microfluidic channels. This article provides a comprehensive look at the physics of the gas solubility and permeability of PDMS, a systematic review of different types of vacuum-driven power-free microfluidics, and guidelines for designing solubility-based or permeability-based PDMS devices, alongside existing applications. Advanced topics and the outlook in using micropumping that utilizes the gas solubility or permeability of PDMS will be also discussed. We strongly recommend that microfluidics and lab-on-chip (LOC) communities harness vacuum energy to develop smart vacuum-driven microfluidic systems.