Polydimethylsiloxane Microfluidics Research Articles

Micropatterned Polymer Nanoarrays with Distinct Superwettability for a Highly Efficient Sweat Collection and Sensing Patch.

Wearable sweat sensor offers a promising means for noninvasive real-time health monitoring, but the efficient collection and accurate analysis of sweat remains challenging. One of the obstacles is to precisely modulate the surface wettability of the microfluidics to achieve efficient sweat collection. Here a facile initiated chemical vapor deposition (iCVD) method is presented to grow and pattern polymer nanocone arrays with distinct superwettability on polydimethylsiloxane microfluidics, which facilitate highly efficient sweat transportation and collection. The nanoarray is synthesized by manipulating monomer supersaturation during iCVD to induce controlled nucleation and preferential vertical growth of fluorinated polymer. Subsequent selective vapor deposition of a conformal hydrogel nanolayer results in superhydrophilic nanoarray floor and walls within the microchannel that provide a large capillary force and a superhydrophobic ceiling that drastically reduces flow friction, enabling rapid sweat transport along varied flow directions. A carbon/hydrogel/enzyme nanocomposite electrode is then fabricated by sequential deposition of highly porous carbon nanoparticles and hydrogel nanocoating to achieve sensitive and stable sweat detection. Further encapsulation of the assembled sweatsensing patch with superhydrophobic nanoarray imparts self-cleaning and water-proof capability. Finally, the sweat sensing patch demonstrates selective and sensitive glucose and lactate detection during the on-body test.

Evaluation of the passive permeability of antidepressants through pore-suspended lipid bilayer

HypothesisThe antidepressant drug imipramine, and its metabolite desipramine show different extents of interaction with, and passive permeation through, cellular membrane models, with the effects depending on the membrane composition. Through multimodal interrogation, we can observe that the drugs have a direct impact on the physicochemical properties of the membrane, that may play a role in their pharmacokinetics. ExperimentsMicrocavity pore-suspended lipid bilayers (MSLBs) of four different compositions, each with a different headgroup charge namely; zwitterionic dioleoylphosphatidylcholine (DOPC), mixed DOPC and negatively charged dioleoylphosphatidylglycerol (DOPG) (3:1), mixed DOPC and positively charged dioleoyltrimethylammoniumpropane (DOTAP) (3:1), and with increasing complex composition mimicking blood-brain-barrier (BBB) were prepared on gold and polydimethylsiloxane (PDMS) substrates using a Langmuir-Blodgett-vesicle fusion method. The molecular interaction and permeation of antidepressants, imipramine, and its metabolite desipramine with the lipid bilayers were evaluated using highly sensitive label-free electrochemical impedance spectroscopy (EIS) and surface-enhanced Raman spectroscopy (SERS). Drug-induced membrane packing/fluidity alterations were assessed using fluorescence lifetime imaging (FLIM) and fluorescence lifetime correlation spectroscopy (FLCS) of MSLB over microfluidic PDMS array. FindingsUsing EIS to evaluate in real-time membrane admittance changes, we found that imipramine greatly increases the ion permeability of negatively charged DOPC:DOPG (3:1) membranes. The effect was observed also at neutral (DOPC) and to a lesser extent at positively charged DOPC:DOTAP(3:1) membranes. In contrast, desipramine had a much weaker impact on ion permeability across all bilayer compositions. Temporal capacitance data show that desipramine intercalates at negatively charged membrane thereby increasing the thickness of the membrane. The overall kinetics of the imipramine permeation is higher than that of desipramine. This was confirmed using SERS, which also provides an evaluation of drug passive permeation based on arrival time across the membrane. Using FLCS, we found that imipramine increases the lipid membrane fluidity, whereas desipramine lowers it, with the exception of the negatively charged membrane. A translocation rate pharmacokinetics model was established for the first time at the MSLB platform by real-time monitoring of the variation in membrane resistance of pristine DOPC and blood-brain-barrier (BBB) membrane.

Artificial Intelligence-Aided Massively Parallel Spectroscopy of Freely Diffusing Nanoscale Entities.

Massively parallel spectroscopy (MPS) of many single nanoparticles in an aqueous dispersion is reported. As a model system, bioconjugated photon-upconversion nanoparticles (UCNPs) with a near-infrared excitation are prepared. The UCNPs are doped either with Tm3+ (emission 450 and 802 nm) or Er3+ (emission 554 and 660 nm). These UCNPs are conjugated to biotinylated bovine serum albumin (Tm3+-doped) or streptavidin (Er3+-doped). MPS is correlated with an ensemble spectra measurement, and the limit of detection (1.6 fmol L-1) and the linearity range (4.8 fmol L-1 to 40 pmol L-1) for bioconjugated UCNPs are estimated. MPS is used for observing the bioaffinity clustering of bioconjugated UCNPs. This observation is correlated with a native electrophoresis and bioaffinity assay on a microtiter plate. A competitive MPS bioaffinity assay for biotin is developed and characterized with a limit of detection of 6.6 nmol L-1. MPS from complex biological matrices (cell cultivation medium) is performed without increasing background. The compatibility with polydimethylsiloxane microfluidics is proven by recording MPS from a 30 μm deep microfluidic channel.

Microfluidic integrated D-shaped optical fiber SERS probe with high sensitivity and ability of multi-molecule detection.

In order to enhance the sensitivity, integration, and practical application capability of Raman detection systems, we propose a multi-channel microfluidic integrated D-shaped optical fiber SERS (Surface-enhanced Raman scattering) probe structure. Firstly, a microfluidic polydimethylsiloxane (PDMS) channel was fabricated using a self-designed multi-channel microfluidic template. Secondly, a uniform layer of silver nanoparticles was deposited on the D-shaped optical fiber using the liquid-liquid interface method. Finally, the D-shaped optical fiber was plasma-bonded to the multi-channel microfluidic channel and a cover glass, resulting in a microfluidic integrated D-shaped optical fiber SERS probe. The prepared sample exhibited excellent detection performance for R6G (rhodamine 6 G) with a detection limit as low as 10-11 mol/L and an enhancement factor of 1.14 × 109. Moreover, the multi-channel configuration enables simultaneous detection of multiple molecules, demonstrating excellent multi-channel multiplexing capability.

Sensing High 17β-Estradiol Concentrations Using a Planar Microwave Sensor Integrated with a Microfluidic Channel.

The global issue of pollution caused by endocrine-disrupting chemicals (EDCs) has been gaining increasing attention. Among the EDCs of environmental concern, 17β-estradiol (E2) can produce the strongest estrogenic effects when it enters the organism exogenously through various routes and has the potential to cause harm, including malfunctions of the endocrine system and development of growth and reproductive disorders in humans and animals. Additionally, in humans, supraphysiological levels of E2 have been associated with a range of E2-dependent disorders and cancers. To ensure environmental safety and prevent potential risks of E2 to human and animal health, it is crucial to develop rapid, sensitive, low cost and simple approaches for detecting E2 contamination in the environment. A planar microwave sensor for E2 sensing is presented based on the integration of a microstrip transmission line (TL) loaded with a Peano fractal geometry with a narrow slot complementary split-ring resonator (PF-NSCSRR) and a microfluidic channel. The proposed technique offers a wide linear range for detecting E2, ranging from 0.001 to 10 mM, and can achieve high sensitivity with small sample volumes and simple operation methods. The proposed microwave sensor was validated through simulations and empirical measurements within a frequency range of 0.5-3.5 GHz. The E2 solution was delivered to the sensitive area of the sensor device via a microfluidic polydimethylsiloxane (PDMS) channel with an area of 2.7 mm2 and sample value of 1.37 µL and measured by a proposed sensor. The injection of E2 into the channel resulted in changes in the transmission coefficient (S21) and resonance frequency (Fr), which can be used as an indicator of E2 levels in solution. The maximum quality factor of 114.89 and the maximum sensitivity based on S21 and Fr at a concentration of 0.01 mM were 1746.98 dB/mM and 40 GHz/mM, respectively. Upon comparing the proposed sensor with the original Peano fractal geometry with complementary split-ring (PF-CSRR) sensors without a narrow slot, several parameters were evaluated, including sensitivity, quality factor, operating frequency, active area, and sample volume. The results showed that the proposed sensor exhibited an increased sensitivity of 6.08% and had a 40.72% higher quality factor, while the operating frequency, active area, and sample volume showed decreases of 1.71%, 25%, and 28.27%, respectively. The materials under tests (MUTs) were analyzed and categorized into groups using principal component analysis (PCA) with a K-mean clustering algorithm. The proposed E2 sensor has a compact size and simple structure that can be easily fabricated with low-cost materials. With the small sample volume requirement, fast measurement with a wide dynamic range, and a simple protocol, this proposed sensor can also be applied to measure high E2 levels in environmental, human, and animal samples.

The Additive Manufacturing Approach to Polydimethylsiloxane (PDMS) Microfluidic Devices: Review and Future Directions.

This paper presents a comprehensive review of the literature for fabricating PDMS microfluidic devices by employing additive manufacturing (AM) processes. AM processes for PDMS microfluidic devices are first classified into (i) the direct printing approach and (ii) the indirect printing approach. The scope of the review covers both approaches, though the focus is on the printed mold approach, which is a kind of the so-called replica mold approach or soft lithography approach. This approach is, in essence, casting PDMS materials with the mold which is printed. The paper also includes our on-going effort on the printed mold approach. The main contribution of this paper is the identification of knowledge gaps and elaboration of future work toward closing the knowledge gaps in fabrication of PDMS microfluidic devices. The second contribution is the development of a novel classification of AM processes from design thinking. There is also a contribution in clarifying confusion in the literature regarding the soft lithography technique; this classification has provided a consistent ontology in the sub-field of the fabrication of microfluidic devices involving AM processes.

Ultraviolet control of bacterial biofilms in microfluidic chips.

Polydimethylsiloxane (PDMS) microfluidic systems have been instrumental in better understanding couplings between physical mechanisms and bacterial biofilm processes, such as hydrodynamic effects. However, precise control of the growth conditions, for example, the initial distribution of cells on the substrate or the boundary conditions in a flow system, has remained challenging. Furthermore, undesired bacterial colonization in crucial parts of the systems, in particular, in mixing zones or tubing, is an important factor that strongly limits the duration of the experiments and, therefore, impedes our ability to study the biophysics of biofilm evolving over long periods of time, as found in the environment, in engineering, or in medicine. Here, we develop a new approach that uses ultraviolet-C (UV-C) light-emitting diodes (LEDs) to confine bacterial development to specific zones of interest in the flow channels. The LEDs are integrated into a 3D printed light guide that is positioned upon the chip and used to irradiate germicidal UV-C directly through the PDMS. We first demonstrate that this system is successful in controlling undesired growth of Pseudomonas aeruginosa biofilm in inlet and outlet mixing zones during 48 h. We further illustrate how this can be used to define the initial distribution of bacteria to perturb already formed biofilms during an experiment and to control colonization for seven days-and possibly longer periods of time-therefore opening the way toward long-term biofilm experiments in microfluidic devices. Our approach is easily generalizable to existing devices at low cost and may, thus, become a standard in biofilm experiments in PDMS microfluidics.

Impacts of Liquid Level on Microwave Resonance Sensing with a Flexible Microfluidic Channel

AbstractPermittivity sensing based on resonance tracking lays the fundamental principle for a variety of biochemical sensors, which has found vast applications in cancer biomarker detection, antigen‐antibody analysis, and so on. Driven by continuous promotion of the detection limit, precise environmental control highlights its critical importance. Here, the impacts of liquid level on microwave resonance sensing are investigated, in which a flexible polydimethylsiloxane microfluidic channel and soft tubing are employed to control the liquid under test. The hydraulic pressure affects the effective permittivity of the liquid and the channel material, hence causing extra resonance shift signals. Both contactless and contacting sensing scenarios are studied in numerical simulations and experiments. It is demonstrated that the resonance frequency varies sensitively with the liquid level, and a sensitivity of 343 kHz mm−1 is measured. Meanwhile, a spoof localized surface plasmon resonator and its optimized excitation structure are employed and analyzed for a good figure of merit, addressing the detectability difficulties for high‐permittivity and high‐loss aqueous solutions. These results provide general guidelines for understanding and controlling the resonance sensors in aqueous environments and help to realize further lower detection limits.

Microfluidic cell engineering on high-density microelectrode arrays for assessing structure-function relationships in living neuronal networks.

Neuronal networks in dissociated culture combined with cell engineering technology offer a pivotal platform to constructively explore the relationship between structure and function in living neuronal networks. Here, we fabricated defined neuronal networks possessing a modular architecture on high-density microelectrode arrays (HD-MEAs), a state-of-the-art electrophysiological tool for recording neural activity with high spatial and temporal resolutions. We first established a surface coating protocol using a cell-permissive hydrogel to stably attach a polydimethylsiloxane microfluidic film on the HD-MEA. We then recorded the spontaneous neural activity of the engineered neuronal network, which revealed an important portrait of the engineered neuronal network-modular architecture enhances functional complexity by reducing the excessive neural correlation between spatially segregated modules. The results of this study highlight the impact of HD-MEA recordings combined with cell engineering technologies as a novel tool in neuroscience to constructively assess the structure-function relationships in neuronal networks.

Leveraging the elastic deformability of polydimethylsiloxane microfluidic channels for efficient intracellular delivery.

With the rapid development of microfluidic based cell therapeutics systems, the need arises for compact, modular, and microfluidics-compatible intracellular delivery platforms with small footprints and minimal operational requirements. Physical deformation of cells passing through a constriction in a microfluidic channel has been shown to create transient membrane perturbations that allow passive diffusion of materials from the outside to the interior of the cell. This mechanical approach to intracellular delivery is simple to implement and fits the criteria outlined above. However, available microfluidic platforms that operate through this mechanism are traditionally constructed from rigid channels with fixed dimensions that suffer from irreversible clogging and incompatibility with larger size distributions of cells. Here we report a flexible and elastically deformable microfluidic channel, and we leverage this elasticity to dynamically generate temporary constrictions with any given size within the channel width parameters. Additionally, clogging is prevented by increasing the size of the constriction momentarily to allow clogs to pass. By tuning the size of the constriction appropriately, we show the successful delivery of GFP-coding plasmids to the interior of three mammalian cell lines and fluorescent gold nanoparticles to HEK293 FT cells all the while maintaining a high cell viability rate. We also demonstrate the device capabilities by systematically identifying the optimum constriction size that maximizes the intracellular delivery efficiency of FITC-dextran for three different cell lines. This development will no doubt lead to miniaturized intracellular delivery microfluidic components that can be easily integrated into larger lab-on-a-chip systems for future cell modification devices.

An Improved Split-Ring Resonator-Based Sensor for Microfluidic Applications.

This study proposes an ultrahigh-sensitivity split-ring resonator-based microwave sensor for retrieving the complex permittivity of liquid samples. An interdigital capacitor structure was used to expand the sensing area and the sensitivity. A defected ground structure and A parallel dual split-ring resonator were introduced to improve the quality factor. A polydimethylsiloxane microfluidic channel substrate was placed above the interdigital capacitor structure. The channel route coincided with the interdigital gap to fully utilize the strong electric field. Ethanol-water solutions with varying ethanol fractions were injected into the channel as the testing liquid. It was demonstrated that the variation in resonant frequency can be used to retrieve the dielectric properties of liquid samples. The proposed sensor used a small liquid volume of ~0.68 μL and provided values in good agreement with the reference data.

A simple and accessible approach for processing photopolymer master molds for the fabrication of microfluidic polydimethylsiloxane devices

The use of three-dimensional (3D) printing for fabrication of master molds for microfluidic devices is very attractive due to its availability and simplicity and replaces the standard methods of soft lithography. However, the commercially available photopolymer resins inhibit the curing of polydimethylsiloxane (PDMS), preventing reliable replication of 3D printed master mold structures. Here, we present a simple and safe method to post-process 3D printed photopolymer master molds for PDMS microfluidic devices. This approach expands the possibilities of prototyping microfluidic PDMS devices for a wider research community without complex post-processing tools currently required for fabrication of 3D photopolymer master molds.

Microfabrication Bonding Process Optimization for a 3D Multi-Layer PDMS Suspended Microfluidics

Microfluidic systems have received increased attention due to their wide variety of applications, from chemical sensing to biological detection to medical analysis. Microfluidics used to be fabricated by using etching techniques that required cleanroom and aggressive chemicals. However, another microfluidic fabrication technique, namely, soft lithography, is less expensive and safer compared to former techniques. Polydimethylsiloxane (PDMS) has been widely employed as a fabrication material in microfluidics by using soft lithography as it is transparent, soft, bio-compatible, and inexpensive. In this study, a 3D multi-layer PDMS suspended microfluidics fabrication process using soft lithography is presented, along with its manufacturing issues that may deteriorate or compromise the microsystem’s test results. The main issues considered here are bonding strength and trapped air-bubbles, specifically in multi-layer PDMS microfluidics. In this paper, these two issues have been considered and resolved by optimizing curing temperature and air-vent channel integration to a microfluidic platform. Finally, the suspended microfluidic system has been tested in various experiments to prove its sensitivity to different fluids and flow rates.

Aerotaxis Assay in Caenorhabditis elegans to Study Behavioral Plasticity.

C. elegans shows robust and reproducible behavioral responses to oxygen. Specifically, worms prefer O 2 levels of 5-10% and avoid too high or too low O 2 . Their O 2 preference is not fixed but shows plasticity depending on experience, context, or genetic background. We recently showed that this experience-dependent plasticity declines with age, providing a useful behavioral readout for studying the mechanisms of age-related decline of neural plasticity. Here, we describe a technique to visualize behavioral O 2 preference and its plasticity in C. elegans , by creating spatial gradients of [O 2 ] in a microfluidic polydimethylsiloxane (PDMS) chamber and recording the resulting spatial distribution of the animals.

Flexible liquid metal display using 3-Aminopropyl triethoxysilane-treated light emitting diodes (LEDs) array

In recent years, the fabrication of flexible electronic devices has been focused on the use eutectic gallium‑indium (EGaIn) liquid metal (LM) materials with polydimethylsiloxane (PDMS) microfluidics, which are less toxic. The flexible electronic devices offer the advantage of attaining various shapes as they are liquid at room temperature. However, discontinuities in the circuit occur while bending of the flexible LM circuit as EGaIn LM easily leaks into the space between the electrical components such as light-emitting diodes (LEDs) and PDMS microfluidic channels. Thus, the present study proposes use of a (3-Aminopropyl) triethoxysilane (APTES)-treatment method to create a circuit consisting of 100 μm channels based on EGaIn. The strategy enhances the bonding strength of surface mount device (SMD) such as LEDs and PDMS. The circuit mold to be patterned with the PDMS has been made with a 3D printer and coated with a polymer as buffer layer to allow the PDMS to detach the circuit mold easily. The change in resistance of the EGaIn LM by varying the bending angle from 0° to 60° is presented for one channel through the I-V curves. Additionally, effect of bending on the on/off state of an APTES-treated SMD type LED has been studied. Furthermore, the proposed method has been employed for developing a 5 × 1 LED array circuit. An Arduino has been employed to express a 5 × 5 LED array circuit in the form of letters. Hence, a flexible device with improved stability has been demonstrated using EGaIn LM that does not overflow while bending due to an enhanced binding force between the PDMS and SMD-type LEDs as a result of the APTES-treatment processing.

Quantitative, Temperature-Calibrated and Real-Time Glucose Biosensor Based on Symmetrical-Meandering-Type Resistor and Intertwined Capacitor Structure.

Here, we propose a glucose biosensor with the advantages of quantification, excellent linearity, temperature-calibration function, and real-time detection based on a resistor and capacitor, in which the resistor works as a temperature sensor and the capacitor works as a biosensor. The resistor has a symmetrical meandering type structure that increases the contact area, leading to variations in resistance and effective temperature monitoring of a glucose solution. The capacitor is designed with an intertwined structure that fully contacts the glucose solution, so that capacitance is sensitively varied, and high sensitivity monitoring can be realized. Moreover, a polydimethylsiloxane microfluidic channel is applied to achieve a fixed shape, a fixed point, and quantitative measurements, which can eliminate influences caused by fluidity, shape, and thickness of the glucose sample. The glucose solution in a temperature range of 25–100 °C is measured with variations of 0.2716 Ω/°C and a linearity response of 0.9993, ensuring that the capacitor sensor can have reference temperature information before detecting the glucose concentration, achieving the purpose of temperature calibration. The proposed capacitor-based biosensor demonstrates sensitivities of 0.413 nF/mg·dL−1, 0.048 nF/mg·dL−1, and 0.011 pF/mg·dL−1; linearity responses of 0.96039, 0.91547, and 0.97835; and response times less than 1 second, respectively, at DC, 1 kHz, and 1 MHz for a glucose solution with a concentration range of 25–1000 mg/dL.

Microfluidic system evaluation for the semi-automatic detection of MOG-IgG in serum samples

Myelin oligodendrocyte glycoprotein reactive immunoglobulin G antibody (MOG-IgG) is in patients with central nervous system demyelination. Reliability of the conventional detection method relies on technician skills and pipetting error accumulation. This work develops a microfluidic system for semi-automatic MOG-IgG detection using cell-based immunofluorescence (IF) assay. The polydimethylsiloxane (PDMS) microfluidic was modified by poly-l-lysine to enhance the adhesion of Human embryonic kidney (HEK) cell. The untransfected and GFP-MOG transfected HEK cells were cultured, fixed, and stained in the microfluidic with the feeding reagents regulated by a syringe pump. Cell characterization, limit of detection (LOD), and turnaround time of the IF assay operation in microfluidic were compared to those in standard microplate. In microfluidic, cell-clumping formation can be avoided and thus signal variations that are caused by cell overlapping can be significantly reduced. LOD of MOG-IgG detection in the microfluidic is at least 2.5 times better than that in the microplate. Signal intensities of the IF staining for 1 h in microfluidic are comparable to those stained for overnight in the standard microplate. By integration with a serial dilution microfluidic, the optimal cutoff titer for MOG-IgG positivity in the patient samples was determined by Receiver operating characteristic curve (ROC) analysis.

Ultrahigh-Sensitivity Microwave Microfluidic Sensors Based on Modified Complementary Electric-LC and Split-Ring Resonator Structures

The paper proposes an ultrahigh sensitivity microwave sensor for retrieving the complex permittivity of the liquid sample. The proposed sensor originally evolves from the complementary split-ring resonator-based sensor, which confines the electric field in the slot of the resonator. A complementary electric-LC resonator structure is applied to substitute the CSRR due to its excellent ability of concentrating the electric field. To further improve the field confinement, a meander slot is embedded in the central arm of the resonator. Then, two etched complementary split-ring resonators are added on both sides of the central meander arm of the modified complementary electric-LC resonator. A polydimethylsiloxane microfluidic channel substrate is placed on the meander arm, and a mixture of water-ethanol is injected. The variations in the resonant frequency and peak attenuation are used to retrieve the complex permittivity of the injected liquid sample. The developed sensors are fabricated and tested, and a good agreement is observed between measured data and reference values. The peak sensitivities in retrieving the relative permittivity achieve 2.5% and 2.71% with the proposed sensors, which are much higher than previously reported designs.

Biocompatible Micron-Scale Silk Fibers Fabricated by Microfluidic Wet Spinning.

For successful material deployment in tissue engineering, the material itself, its mechanical properties, and the microscopic geometry of the product are of particular interest. While silk is a widely applied protein-based tissue engineering material with strong mechanical properties, the size and shape of artificially spun silk fibers are limited by existing processes. This study adjusts a microfluidic spinneret to manufacture micron-sized wet-spun fibers with three different materials enabling diverse geometries for tissue engineering applications. The spinneret is direct laser written (DLW) inside a microfluidic polydimethylsiloxane (PDMS) chip using two-photon lithography, applying a novel surface treatment that enables a tight print-channel sealing. Alginate, polyacrylonitrile, and silk fibers with diameters down to 1µm are spun, while the spinneret geometry controls the shape of the silk fiber, and the spinning process tailors the mechanical property. Cell-cultivation experiments affirm bio-compatibility and showcase an interplay between the cell-sized fibers and cells. The presented spinning process pushes the boundaries of fiber fabrication toward smaller diameters and more complex shapes with increased surface-to-volume ratio and will substantially contribute to future tailored tissue engineering materials for healthcareapplications.

Easy Surface Functionalization and Bioconjugation of Peptides as Capture Agents of a Microfluidic Biosensing Platform for Multiplex Assay in Serum.

The development of assays for protein biomarkers in complex matrices is a demanding task that still needs implementation of new approaches. Antibodies as capture agents have been largely used in bioassays but their low stability, low-efficiency production, and cross-reactivity in multiplex approaches impairs their larger applications. Instead, synthetic peptides, even with higher stability and easily adapted amino acid sequences, still remain largely unexplored in this field. Here, we provide a proof-of-concept of a microfluidic device for direct detection of biomarker overexpression. The multichannel microfluidic polydimethylsiloxane (PDMS) device was first derivatized with PAA (poly(acrylic acid)) solution. CRP-1, VEGF-114, and ΦG6 peptides were preliminarily tested to respectively bind the biomarkers, C-reactive protein (CRP), vascular endothelial growth factor (VEGF), and tumor necrosis factor-alpha (TNF-α). Each PDMS microchannel was then respectively bioconjugated with a specific peptide (CRP-1, VEGF-114, or ΦG6) to specifically capture CRP, VEGF, and TNF-α. With such microdevices, a fluorescence bioassay has been set up with sensitivity in the nanomolar range, both in buffered solution and in human serum. The proposed multiplex assay worked with a low amount of sample (25 μL) and detected biomarker overexpression (above nM concentration), representing a noninvasive and inexpensive screening platform.