Magnetic particle dosing and size separation in a microfluidic channel

Separation of functional magnetic particles or magnetically labeled entities is a key feature for bioanalytical or biomedical applications and therefore also an important component of lab-on-a-chip devices for biological applications. We present a novel integrated microfluidic magnetic bead manipulation device, comprising dosing of magnetic particles, controlled release and subsequent magnetophoretic size separation with high resolution. The system is designed to meet the requirements of specific bioassays, in particular of on-chip agglutination assays for the detection of rare analytes by particle coupling as doublets. Integrated soft-magnetic microtips with different shapes provide the magnetic driving force of the bead manipulation protocol. The magnetic tips that serve as field concentrators of an external electromagnetic field, are positioned in close contact to a microfluidic channel in order to generate high magnetic actuation forces. Mixtures of 1.0 μm and 2.8 μm superparamagnetic beads have been used to characterize the system. Magnetophoretic size separation with high resolution was performed in static conditions and in continuous flow mode. In particular, we could demonstrate the separation of 1.0 μm single beads and doublets in a sample flow.     

Size-selective separation of magnetic nanospheres in a microfluidic channel

This paper reported an efficient method to size-selective separate magnetic nanospheres using a self-focusing microfluidic channel equipped with a permanent magnet. Under external magnetic field, the magnetophoresis force exerted on particles leads to size-dependent deflections from their laminar flow paths and results in effective particles separation. By adjusting the distance between magnet and main path of channel, we obtained two monodisperse nanosphere samples (Ca. 90 nm, Ca. 160 nm) from polydispersing particles solution whose diameters varied from 40 to 280 nm. Based on the magnetostatic and laminar flow models, numerical simulations were also used to predict and optimize the nanospheres migrations. Two thresholds of particles diameters were obtained by the simulations and diverse at each position of magnet. Therefore, appropriate position of the magnet could be determined at a certain particle sizes’ range when the flow rate of the two inlets remains unchanged.     

A numerical design study of chaotic mixing of magnetic particles in a microfluidic bio-separator

A two-dimensional numerical investigation into the mixing of magnetic microparticles with bio-cells in a chaotic micromixer is carried out by using a multiphysics finite element analysis package. Fluid and magnetic problems are simulated in steady-state and time-dependent modes, respectively. Intensity of segregation is utilized as the main index to examine the efficiency of the mixer. Trajectories of the particles are used in order to detect chaos in their motion and quantify its extent. Moreover, probability of the collision between particles and target bio-cells is examined as a supplemental index to study the effects of driving parameters on the mixing process. Simulation results reveal that while in some ranges of operating conditions all indices are in good agreement, there are some ranges where they appear to predict contradicting results which is discussed in details. It is found that optimum operating conditions for the system is obtained when the Strouhal number is less than 0.6, which corresponds to the efficiency of about 85% in a mixing length of 500 μm (The mixer design described here is patent pending).     

A microfluidic sensor based on ferromagnetic resonance induced in magnetic bead labels

This report details preliminary studies towards the development of a microfluidic sensor that exploits ferromagnetic resonance, excited in magnetic bead labels, for signal transduction. The device consists of a microwave circuit in which a slotline and a coplanar waveguide are integrated with a biochemically activated sensor area. The magnetic beads are immobilized in the sensor area by bio-specific reactions. A microwave signal applied to the slotline is coupled to the coplanar waveguide only in the presence of magnetic beads at the functionalized sensor area. Ferromagnetic resonance in the beads further enhances the coupling. This inductive detection technique lends itself to miniaturization, is inexpensive to fabricate and can be adapted for the detection of a wide range of molecules for which bio-specific ligands are available.Experimentally, the variation of the output signal as a function of the location of magnetic beads was studied for the proposed technique. Subsequently, a prototype device was constructed by biotinylation of the sensor area and integration with a microfluidic chip fabricated in polydimethyl siloxane (PDMS). Preliminary experiments were conducted on this prototype using streptavidin-functionalized magnetic beads as labels. It was shown that the magnetic beads, immobilized at the sensor area by streptavidin-biotin linkage, produced a distinct ferromagnetic resonance response easily discernable from the background signal.     

Acoustically induced bubbles in a microfluidic channel for mixing enhancement

Due to small dimensions and low fluid velocity, mixing in microfluidic systems is usually poor. In this study, we report a method of enhancing microfluidic mixing using acoustically induced gas bubbles. The effect of applied frequency on mixing was investigated over the range 0.5–10 kHz. Under either low frequency 0.5 kHz or high frequency 10 kHz, no noticeable improvement in the present mixer was observed. However, a significant increase in the mixing efficiency was achieved within a window of the frequencies between 1.0 and 5.0 kHz. It was found in our present microfluidic structure, single (or multi-) bubble(s) could be acoustically generated under the frequency ranging from 1.0 to 5.0 kHz by a piezoelectric disc. The interaction between bubble and acoustic field causes bubble oscillation which in turn could disturb local flow field to result in mixing enhancement.     

Continuous generation of hydrogel beads and encapsulation of biological materials using a microfluidic droplet-merging channel

In this paper, we describe a method for encapsulation of biomaterials in hydrogel beads using a microfluidic droplet-merging channel. We devised a double T-junction in a microfluidic channel for alternate injection of aqueous fluids inside a droplet unit carried within immiscible oil. With this device, hydrogel beads with diameter <100 μm are produced, and various solutions containing cells, proteins and reagents for gelation could merge with the gel droplets with high efficiency in the broad range of flow rates. Mixing of reagents and reactions inside the hydrogel beads are continuously observed in a microchannel through a microscope. By enabling serial injection of each liquid with the dispersed gel droplets after they are produced from the oil-focusing channel, the device simplifies the sample preparation process, and gel-bead fabrication can be coupled with further assay continuously in a single channel. Instantaneous reactions of enzyme inside hydrogel and in-situ formation of cell-containing beads with high viability are demonstrated in this report.     

Dynamic coating for protein separation in cyclic olefin copolymer microfluidic devices

We report our study on using hydroxyethyl cellulose (HEC) as a dynamic coating for protein separation in microfluidic devices made from cyclic olefin copolymer (COC). The coating significantly enhances hydrophilicity of COC surface, evident from the decrease in contact angle of water in a COC channel. Surface treatment of COC channels with HEC also results in a 72% drop in electroosmotic (EO) mobility and a significant reduction in protein adsorption on the channel wall. Using bovine serum albumin as a model protein, the number of theoretical plates of 1.1 × 10⁴ was achieved in a separation distance of 3.3 cm using free solution electrophoresis. Hydroxyethyl cellulose dynamic coating is also found to have an effect on isoelectric focusing (IEF) of proteins. It not only prevents proteins from adsorption, but also reduces EO flow, both of which help achieve IEF of proteins with a difference of 0.1 pH values in isoelectric points (pI).     

New design for the separation of microorganisms using microfluidic deterministic lateral displacement

This paper presents a new design of microfluidic device for the separation of soft microorganisms using the deterministic lateral displacement principle. The novelty of the proposed design is based on creating new post shape that should address the challenges posed using the conventional circular post arrays. Based on experimental observations of trypanosome particles, circular posts have negative effects on the separation process of deformable and non-spherical cells. These effects reside in either shifting soft cells toward random streamlines further down the array or clogging the device. Airfoil and diamond post shapes have been proposed and compared against the circular posts. The flow around these posts has been simulated and analyzed numerically based on computational fluid dynamic (CFD). The results show that airfoil post shape has the potential to overcome complications when working with soft biological samples, namely the cells that are not rigid and not spherical. Microfluidic systems based on airfoil obstacles could easily be manufactured for clinical laboratory and biomedical applications providing a good promise in nano/micro-separation field.     

Computational fluid dynamics analysis of microbubble formation in microfluidic flow-focusing devices

Bubble formation in a microfluidic flow-focusing device is simulated using the volume-of-fluid approach to achieve a complete solution of the Navier–Stokes equations for both the gas and liquid phases. The results of the simulation show good agreement with previous experimental results. A detailed examination of the predicted pressure and velocity profiles from the simulation also provide further validation for the conclusions drawn previously with experimental results. The simulation results show the existence of two distinct modes of bubble formation. Simulations of systems an order of magnitude smaller than those investigated experimentally indicate that such reduced systems sizes are a viable approach that would result in much smaller bubble sizes.     

Fabrication of microfluidic device channel using a photopolymer for colloidal particle separation

We have developed a method of fabricating microfluidic device channels for bio-nanoelectronics system by using high performance epoxy based dry photopolymer films or dry film resists (DFRs). The DFR used was with a trademark name Ordyl SY355 from Elga Europe. The developing and exposing processes as well as the time taken in making the channels are recorded. Finally from those recorded methods, the accurate procedures and time taken for DFR development and exposure have been found and ultimately been consistently used in fabricating our channels. These channels were patterned and sandwiched in between two glass substrates. In our advance, the channel was formed for the colloidal particle separation system. They can be used for handling continuous fluid flow and particle repositioning maneuver using dielectrophoresis that have showed successful results in the separation.     

Characterization of passive microfluidic mixers fabricated using soft lithography

This work characterizes microfluidic mixers fabricated using soft lithography without grooves or with grooves on the top and/or bottom of the channels. The purpose of this study was to investigate whether grooves on the top and bottom of the channel significantly improve mixing in microfluidic systems. The channels studied were 200 μm wide with repeating sets of alternating patterns of diagonal stripes and chevrons. The study employed confocal microscopy to investigate the mixing of a 0.1 wt% fluorescein solution with a deionized water solution. The results of the study indicate a 10% improvement in mixing over systems with grooves only on the top of the channel.     

Theoretical analysis of a new, efficient microfluidic magnetic bead separator based on magnetic structures on multiple length scales

We present a theoretical analysis of a new design for microfluidic magnetic bead separation. It combines an external array of mm-sized permanent magnets with magnetization directions alternating between up and down with μm-sized soft magnetic structures integrated in the bottom of the separation channel. The concept is studied analytically for simple representative geometries and by numerical simulation of an experimentally realistic system geometry. The array of permanent magnets provides long-range magnetic forces that attract the beads to the channel bottom, while the soft magnetic elements provide strong local retaining forces that prevent captured beads from being torn loose by the fluid drag. The addition of the soft magnetic elements increases the maximum retaining force by two orders of magnitude. The design is scalable and provides an efficient and simple solution to the capture of large amounts of magnetic beads on a microsystem platform.     

Design optimization of an enzymatic assay in an electrokinetically-driven microfluidic device

Microfluidic systems are increasingly popular for rapid and cheap determinations of enzyme assays and other biochemical analysis. In this study reduced order models (ROM) were developed for the optimization of enzymatic assays performed in a microchip. The model enzyme assay used was β-galactosidase (β-Gal) that catalyzes the conversion of Resorufin β-d-galactopyranoside (RBG) to a fluorescent product as previously reported by Hadd et al. (Anal Chem 69(17): 3407–3412, 1997). The assay was implemented in a microfluidic device as a continuous flow system controlled electrokinetically and with a fluorescence detection device. The results from ROM agreed well with both computational fluid dynamic (CFD) simulations and experimental values. While the CFD model allowed for assessment of local transport phenomena, the CPU time was significantly reduced by the ROM approach. The operational parameters of the assay were optimized using the validated ROM to significantly reduce the amount of reagents consumed and the total biochip assay time. After optimization the analysis time would be reduced from 20 to 5.25 min which would also resulted in 50% reduction in reagent consumption.     

A rapid polymerisation process realizing 3D biocompatible structures in a microfluidic channel suitable for genetic analysis

Surface probe immobilisation is a complex and time consuming task undertaken prior to microfluidic integration, this requires surface functionalisation, biomolecule spotting, incubation and blocking steps. Traditional bonding techniques (anodic, thermal, etc.) or adhesives (UV cured) used to seal fluidic systems may denature biomolecules due to high temperature or vapour effects, thus bonding techniques such as thin film laminate or PDMS are used to seal systems, with substrate-fluidic alignment required prior to bonding. We propose a technique allowing probe DNA molecules to be immobilised in a sealed microfluidic system using (3D) hydrogel structures without any alignment steps. A prepolymer solution is introduced to the channels where photo-polymerisation is undertaken forming 3D structures covalently attached to the channel surface. We use a photo-initiated prepolymer material poly-ethylene-glycol (PEG) to form structures containing probe DNA. This process is fast compared to conventional biomolecule immobilisation techniques and is also biocompatible, this direct write approach removes overnight immobilisation/incubation of the probe DNA, it also facilitates immobilisation within a sealed fluidic system where conventionally DNA probe spots must be immobilised prior to channel sealing. We consider the transport of target DNA from bulk analyte to the 3D gel structure and evaluate hybridisation within the microfluidic system.     

Towards a continuous microfluidic rheometer

In a previous paper we presented a way to measure the rheological properties of complex fluids on a microfluidic chip (Guillot et al., Langmuir 22:6438, 2006). The principle of our method is to use parallel flows between two immiscible fluids as a pressure sensor. In fact, in a such flow, both fluids flow side by side and the size occupied by each fluid stream depends only on both flow rates and on both viscosities. We use this property to measure the viscosity of one fluid knowing the viscosity of the other one, both flow rates and the relative size of both streams in a cross-section. We showed that using a less viscous fluid as a reference fluid allows to define a mean shear rate with a low standard deviation in the other fluid. This method allows us to measure the flow curve of a fluid with less than 250 μL of fluid. In this paper we implement this principle in a fully automated set up which controls the flow rate, analyzes the picture and calculates the mean shear rate and the viscosity of the studied fluid. We present results obtained for Newtonian fluids and complex fluids using this set up and we compare our data with cone and plate rheometer measurements. By adding a mixing stage in the fluidic network we show how this set up can be used to characterize in a continuous way the evolution of the rheological properties as a function of the formulation composition. We illustrate this by measuring the rheological curve of four formulations of polyethylene oxide solution with only 1.3 mL of concentrated polyethylene oxide solution. This method could be very useful in screening processes where the viscosity range and the behavior of the fluid to an applied stress must be evaluated.     

Continuous-flow in-droplet magnetic particle separation in a droplet-based microfluidic platform

This paper presents a continuous-flow in-droplet magnetic particle separation in a droplet-based microfluidic device for magnetic bead-based bioassays. Two functions, electrocoalescence and magnetic particle manipulation, are performed in this device. A pair of charging metallic needles is inserted into two aqueous channels of the device. By electrostatic force, two different solutions can be merged to be mixed at a junction of droplet generation. The manipulation of magnetic particles is achieved using an externally applied magnetic field. The magnetic particles are separated by the magnetic field to one side of the droplet and extracted by splitting the droplet into two daughter droplets: one contains the majority of the magnetic particles and the other is almost devoid of magnetic particles. The applicability of the continuous-flow in-droplet magnetic particle separation is demonstrated by performing a proof-of-concept immunoassay between streptavidin-coated magnetic beads and biotin labelled with fluorescence. This approach will be useful for various biological and chemical analyses and compartmentalization of small samples.     

Electrospinning of hollow and core/sheath nanofibers using a microfluidic manifold

In this article, we describe a microfluidic approach to fabricate hollow and core/sheath nanofibers by electrospinning. Key benefits in using microfluidic devices for nanofiber synthesis include rapid prototyping, ease of fabrication, and the ability to spin multiple fibers in parallel through arrays of individual microchannels. Hollow poly (vinylpyrrolidone) (PVP) + titania (TiO₂) composite and core/sheath polypyrrole (PPy)/PVP nanofibers of the order of 100 and 250 nm, respectively, were successfully fabricated using elastomeric microfluidic devices. Fiber characterization was subsequently carried out using a combination of scanning electron microscopy and transmission electron microscopy.     

Effect of channel geometry on solute dispersion in pressure-driven microfluidic systems

Pressure-driven transport of fluid and solute samples is often desirable in microfluidic devices, particularly where sufficient electroosmotic flow rates cannot be realized or the use of an electric field is restricted. Unfortunately, this mode of actuation also leads to hydrodynamic dispersion due to the inherent fluid shear in the system. While such dispersivity is known to scale with the square of the Peclet number based on the narrower dimension of the conduit (often the channel depth), the proportionality constant can vary significantly depending on its actual cross section. In this article, we review previous studies to understand the effect of commonly microfabricated channel cross sections on the Taylor–Aris dispersion of solute slugs in simple pressure-driven flow systems. We also analyze some recently proposed optimum designs which can reduce the contribution to this band broadening arising from the presence of the channel sidewalls. Finally, new simulation results have been presented in the last section of this paper which describe solutal spreading due to bowing of microchannels that can occur from stresses developed during their fabrication or operation under high-pressure conditions.     

Development of high throughput microfluidic cell culture chip for perfusion 3-dimensional cell culture-based chemosensitivity assay

This study reports a microfluidic cell culture chip encompassing 36 microbioreactors for high throughput perfusion 3-dimensional (3D) cell culture-based chemosensitivity assays. Its advantages include the capability for multiplexed medium delivery, and the function for both efficient and high throughput micro-scale 3D culture construct preparation and loading. The results showed that the proposed medium pumping mechanism was able to provide a uniform pumping rates ranging from 1.2 to 3.9 μl h⁻¹. In addition, the simple cell/hydrogel loading scheme has been proven to be able to carry out 3D cell culture construct preparation and loading precisely and efficiently. Furthermore, a chemosensitivity assay was successfully demonstrated using the proposed cell culture chip. The results obtained were also compared with the same evaluation based on a conventional 2D monolayer cell culture. It can be concluded that the choice of cell culture format can result in different chemosensitivity evaluation results. Overall, because of the nature of miniaturized perfusion 3D cell culture, the cell culture chip not only can provide stable, well-defined and more biologically relevant culture environments, but it also features low consumption of research resources. All these traits are found particularly useful for high-precision and high-throughput 3D cell culture-based assays.