Multiparametric, Label-Free Analysis of Microfluidic Droplets via a Dynamic Phase Grating Approach.

Droplet-based microfluidic systems possess many fundamental advantages as a platform for the analysis of chemical and biological species. However, whereas on-chip operations have rapidly developed over the past decades, approaches for analyzing target molecules within droplets have largely remained limited to methods requiring bulky and expensive instrumentation. In this work, we describe a droplet analysis approach whereby the droplet train itself is the sensing construct. Specifically, the droplet train is interrogated as a transmission phase grating, allowing high-throughput, label-free, solution-phase, and multi-parametric analysis of droplet contents. Importantly, three distinct properties of generated droplets can be simultaneously extracted using this conceptually simple and experimentally straightforward measurement approach. Under constant droplet generation conditions, measurement of droplet viscosity is achieved by monitoring changes in zero order to first order peak separation in the far-field diffraction pattern, with a sensitivity of 2.28 × 10-4 cSt per µm change in peak separation. In parallel, measurement of droplet refractive index (RI) is achieved by measuring changes in the ratio of the zero order to first order peak intensity, with a sensitivity of 2.14 × 10-4 RI units per unit change in a diffracted peak intensity ratio. Finally, droplet generation frequency is determined from the time-varying oscillation of the peak height ratio, yielding comparable results to an expensive high-speed camera commonly used for droplet imaging. Importantly, the experimental strategy for this approach is straightforward and does not require expensive instrumentation; therefore, it may find utility in affordable and portable analysis approaches applied to diverse droplet microfluidic assays.

Characterization of the impact of mixing and droplet volumes on the behavior of microfluidic ion-selective droptodes.

Droplet microfluidic optodes, or "droptodes", have emerged as a powerful technology for rapid detection of small ions in complex matrices. While using segmented aqueous phases provides the benefits of sample isolation, the influence of the liquid nature of the oil carrier phase has not yet been explored. In this paper, we examine the influence of microfluidic parameters on droptode efficiency, using potassium-sensitive droptodes as a model system. We found that while changing flow rates on device does not change droptode performance, both channel geometry and droplet size significantly impact droptode efficiency. Specifically, enhanced mixing of the droplets leads to faster equilibration on device and lowers limits of detection by about one order of magnitude. We also found that increasing the size of the sample droplet, at the expense of the size of the oil carrier/sensing phase, leads to higher sensitivity in the linear region of the droptode. These easily manipulated properties will allow one device to potentially be adapted for several different applications, based upon the type and concentration range of measurement required.

Silicon Photonic Microring Resonator Arrays as a Universal Detector for Capillary Electrophoresis.

Electrophoretic separations conventionally rely on chromogenic, fluorogenic, or redox properties for analyte detection that, in many instances, involve chemical modification of samples prior to analysis. For analytes natively lacking chemical signatures, refractive index-based measurements are appealing as a method to detect these molecules without pretreatment. Microring resonators are a type of whispering gallery mode sensor capable of detecting bulk changes in refractive index. Here, we demonstrate the use of silicon photonic microring resonator arrays as a postcolumn detector for capillary electrophoresis. In this approach, we establish the universal detection capabilities of microrings through calibration with analytes lacking unique spectral signatures. Separations of small molecule mixtures are demonstrated using capillary zone electrophoresis. For these separations, the microring resonators maintain a linear response over several orders of magnitude in concentration for three candidate small molecules. Successful separation of three sugars with direct detection is also demonstrated. We further present the successful separation and detection of three model proteins, exemplifying the promise of microring resonators arrays as a biocompatible detector for capillary electrophoresis. Additionally, the spatially offset, array-based nature of the sensing platform enables real-time analysis of analyte mobility and performance characterization-a combination that is not typically provided using single-point detectors.

Translational Opportunities for Microfluidic Technologies to Enable Precision Epigenomics.

Personalizing health care by taking genetic, environmental, and lifestyle factors into account is central to modern medicine. The crucial and pervasive roles epigenetic factors play in shaping gene-environment interactions are now well recognized. However, identifying robust epigenetic biomarkers and translating them to clinical tests has been difficult due in part to limitations of available platforms to detect epigenetic features genome-wide (epigenomic assays). This Feature introduces several important prospects for precision epigenomics, highlights capabilities and limitations of current laboratory technologies, and emphasizes opportunities for microfluidic tools to facilitate translation of epigenetic analyses to the clinic, with a particular focus on methods to profile gene-associated histone modifications and their impacts on chromatin structure and gene expression.

Coherent silicon photonic interferometric biosensor with an inexpensive laser source for sensitive label-free immunoassays.

Over the past two decades, integrated photonic sensors have been of major interest to the optical biosensor community due to their capability to detect low concentrations of molecules with label-free operation. Among these, interferometric sensors can be read-out with simple, fixed-wavelength laser sources and offer excellent detection limits but can suffer from sensitivity fading when not tuned to their quadrature point. Recently, coherently detected sensors were demonstrated as an attractive alternative to overcome this limitation. Here we show, for the first time, to the best of our knowledge, that this coherent scheme provides sub-nanogram per milliliter limits of detection in C-reactive protein immunoassays and that quasi-balanced optical arm lengths enable operation with inexpensive Fabry-Perot-type lasers sources at telecom wavelengths.

Real-Time Measurement of Polymer Brush Dynamics Using Silicon Photonic Microring Resonators: Analyte Partitioning and Interior Brush Kinetics.

Polymer brushes are found in biomedical and industrial technologies, where they exhibit functionalities considerably dependent on polymer brush-solvent-analyte interactions. It remains a difficult challenge to quickly analyze solvent-swollen polymer brushes, both at the solvent-polymer brush interface and in the brush interior, as well as to monitor the kinetics of interaction of solvent-swollen brushes with key analytes. Here, we demonstrate the novel use of silicon photonic microring resonators to characterize in situ swollen polymer brush-analyte interactions. By monitoring resonant wavelength shifts, we find that brush-solvent-analyte interaction parameters can be extracted from a single set of data or from successive analyte introductions using a single brush-coated sensor. The partition coefficient of three industrially relevant plasticizers into hydrophobic and hydrophilic brushes was determined and found to be in agreement with known solubility trends. We found that the diffusion coefficient of the plasticizer into the brush decreases as brush thickness increases, supporting a model of a dense inner brush layer and diffuse outer layer. pKa's of pH-sensitive brushes were determined on the microring resonator platform; upon increasing the dry brush thickness, the pKa for poly(2-dimethylamino ethyl methacrylate) decreased from 8.5 to approach the bulk material pKa of 7.3 and showed dependence on the presence and concentration of salt. These proof-of-concept experiments show how the surface-sensitive nature of the microring resonator detection platform provides valuable information about the interaction of the polymer brushes with the solvents and analytes, not easily accessed by other techniques.

A linear mass concentration detector for solvent gradient polymer separations.

Characterization of copolymers requires the measurement of two distributions-molecular weight (MW) and chemical composition (CC). Molecular weight distributions (MWD) are traditionally determined using size exclusion chromatography (SEC) run under isocratic solvent conditions. Chemical composition distributions (CCD) are often determined using liquid adsorption chromatography (LC) with solvent gradients. The use of solvent gradients, however, often limits options of compatible detectors. A gradient compatible, universal linear mass concentration detector is a longstanding unmet need. Many industrially-relevant polymers lack chromophores or other discriminating moieties requiring detectors with a universal response. Differential refractive index (dRI) is incompatible with gradient elution due to its small dynamic range. Charged aerosol detectors (CAD) and evaporative light scattering detectors (ELSD) are probably the most promising options for gradient elution detection, but both suffer from a nonlinear mass concentration response. Silicon photonic microring resonators are optical sensors that are responsive to changes in the local refractive index (RI). The substantial dynamic range of this technology makes it attractive for refractive index-based detection during solvent gradient elution. Previously, the microring resonator platform was used as a SEC detector to characterize the MWD of broadly dispersed polystyrene (PS) standards. In this study, we demonstrate the gradient compatibility of the microring resonator platform for polymer detection by quantifying the CCD of polymer blend components. Control experiments were run with UV and ELSD detection, highlighting the uniqueness of the platform as a linear mass concentration detector with a universal detector response.

Impact of Silanization Parameters and Antibody Immobilization Strategy on Binding Capacity of Photonic Ring Resonators.

Ring resonator-based biosensors have found widespread application as the transducing principle in "lab-on-a-chip" platforms due to their sensitivity, small size and support for multiplexed sensing. Their sensitivity is, however, not inherently selective towards biomarkers, and surface functionalization of the sensors is key in transforming the sensitivity to be specific for a particular biomarker. There is currently no consensus on process parameters for optimized functionalization of these sensors. Moreover, the procedures are typically optimized on flat silicon oxide substrates as test systems prior to applying the procedure to the actual sensor. Here we present what is, to our knowledge, the first comparison of optimization of silanization on flat silicon oxide substrates to results of protein capture on sensors where all parameters of two conjugation protocols are tested on both platforms. The conjugation protocols differed in the chosen silanization solvents and protein immobilization strategy. The data show that selection of acetic acid as the solvent in the silanization step generally yields a higher protein binding capacity for C-reactive protein (CRP) onto anti-CRP functionalized ring resonator sensors than using ethanol as the solvent. Furthermore, using the BS3 linker resulted in more consistent protein binding capacity across the silanization parameters tested. Overall, the data indicate that selection of parameters in the silanization and immobilization protocols harbor potential for improved biosensor binding capacity and should therefore be included as an essential part of the biosensor development process.

Silicon Photonic Microring Resonator Arrays for Mass Concentration Detection of Polymers in Isocratic Separations.

Molecular weight distribution (MWD) is often the most informative analytical parameter in polymer analysis, with gel permeation chromatography (GPC) being the most common approach for determining the MWD for polymer samples. Many industrially relevant polymers lack chromogenic or fluorogenic signatures, precluding use of spectroscopy-based detection. Universal detectors, such as evaporative light scattering and charged aerosol detectors, are nonlinear, limiting quantitative polymer analysis. Differential refractive index (dRI) detectors show linear mass concentration sensitivity but are limited for some analyses given that they are incompatible with gradient-based separations, have a limited dynamic range, and require extended thermal equilibration times. In this study, we investigated the utility of silicon photonic microring resonator arrays as a quantitative mass concentration detector for industrial polymer analysis. Microring resonators have optical properties that are sensitive to changes in refractive index, offer an extended dynamic range, have a broad solvent compatibility, and have a linear mass concentration detection for a range of molecular weights. Linear mass concentration detection for microrings was demonstrated through a series of isocratic GPC separations using narrow MWD polystyrene (PS) standards. This detection technology was then utilized in conjunction with conventional GPC detectors to analyze a series of broad MWD PS standards, with results in good agreement with dRI and UV/visible. These results demonstrate the potential of the microring resonator platform as a detector for industrial polymer analysis.

Ionophore-Based Biphasic Chemical Sensing in Droplet Microfluidics.

Droplet microfluidics is an enabling platform for high-throughput screens, single-cell studies, low-volume chemical diagnostics, and microscale material syntheses. Analytical methods for real-time and in situ detection of chemicals in the droplets will benefit these applications, but they remain limited. Reported herein is a novel heterogeneous chemical sensing strategy based on functionalization of the oil phase with rationally combined sensing reagents. Sub-nanoliter oil segments containing pH-sensitive fluorophores, ionophores, and ion-exchangers enable highly selective and rapid fluorescence detection of physiologically important electrolytes (K+ , Na+ , and Cl- ) and polyions (protamine) in sub-nanoliter aqueous droplets. Electrolyte analysis in whole blood is demonstrated without suffering from optical interference from the sample matrix. Moreover, an oil phase doped with an aza-BODIPY dye allows indication of H2 O2 in the aqueous droplets, exemplifying sensing of targets beyond ionic species.