Joshua A. Hagen

Processing techniques for deoxyribonucleic acid: Biopolymer for photonics applications

Marine-based deoxyribonucleic acid (DNA), purified from waste products of the Japanese fishing industry, has recently become a material of interest in photonics applications. Using highly purified DNA, unique processing techniques developed specifically to transform the purified DNA into a biopolymer suitable for optical device fabrication are reported.

Adhesive RFID Sensor Patch for Monitoring of Sweat Electrolytes

Wearable digital health devices are dominantly found in rigid form factors such as bracelets and pucks. An adhesive radio-frequency identification (RFID) sensor bandage (patch) is reported, which can be made completely intimate with human skin, a distinct advantage for chronological monitoring of biomarkers in sweat. In this demonstration, a commercial RFID chip is adapted with minimum components to allow potentiometric sensing of solutes in sweat, and surface temperature, as read by an Android smartphone app with 96% accuracy at 50 mM Na+ (in vitro tests). All circuitry is solder-reflow integrated on a standard Cu/polyimide flexible-electronic layer including an antenna, but while also allowing electroplating for simple integration of exotic metals for sensing electrodes. Optional paper microfluidics wick sweat from a sweat porous adhesive allowing flow to the sensor, or the sensor can be directly contacted to the skin. The wearability of the patch has been demonstrated for up to seven days, and includes a protective textile which provides a feel and appearance similar to a standard Band-Aid. Applications include hydration monitoring, but the basic capability is extendable to other mM ionic solutes in sweat (Cl-, K+, Mg2+, NH4+, and Zn2+). The design and fabrication of the patch are provided in full detail, as the basic components could be useful in the design of other wearable sensors.

The microfluidics of the eccrine sweat gland, including biomarker partitioning, transport, and biosensing implications.

Non-invasive and accurate access of biomarkers remains a holy grail of the biomedical community. Human eccrine sweat is a surprisingly biomarker-rich fluid which is gaining increasing attention. This is especially true in applications of continuous bio-monitoring where other biofluids prove more challenging, if not impossible. However, much confusion on the topic exists as the microfluidics of the eccrine sweat gland has never been comprehensively presented and models of biomarker partitioning into sweat are either underdeveloped and/or highly scattered across literature. Reported here are microfluidic models for eccrine sweat generation and flow which are coupled with review of blood-to-sweat biomarker partition pathways, therefore providing insights such as how biomarker concentration changes with sweat flow rate. Additionally, it is shown that both flow rate and biomarker diffusion determine the effective sampling rate of biomarkers at the skin surface (chronological resolution). The discussion covers a broad class of biomarkers including ions (Na(+), Cl(-), K(+), NH4 (+)), small molecules (ethanol, cortisol, urea, and lactate), and even peptides or small proteins (neuropeptides and cytokines). The models are not meant to be exhaustive for all biomarkers, yet collectively serve as a foundational guide for further development of sweat-based diagnostics and for those beginning exploration of new biomarker opportunities in sweat.

Optimization of a Paper-Based ELISA for a Human Performance Biomarker

Monitoring aspects of human performance during various activities has recently become a highly investigated research area. Many new commercial products are available now to monitor human physical activity or responses while performing activities ranging from playing sports, to driving, and even sleeping. However, monitoring cognitive performance biomarkers, such as neuropeptides, is still an emerging field due to the complicated sample collection and processing, as well as the need for a clinical lab to perform analysis. Enzyme-linked immunosorbent assays (ELISAs) provide specific detection of biomolecules with high sensitivity (picomolar concentrations). Even with the advantage of high sensitivity, most ELISAs need to be performed in a laboratory setting and require around 6 h to complete. Transitioning this assay to a platform where it reduces cost, shortens assay time, and is able to be performed outside a lab is invaluable. Recently developed paper diagnostics provide an inexpensive platform on which to perform ELISAs; however, the major limiting factor for moving out of the laboratory environment is the measurement and analysis instrumentation. Using something as simple as a digital camera or camera-enabled Windows- or Android-based tablets, we are able to image paper-based ELISAs (P-ELISAs), perform image analysis, and produce response curves with high correlation to target biomolecule concentration in the 10 pM range. Neuropeptide Y detection was performed. Additionally, silver enhancement of Au NPs conjugated with IgG antibodies showed a concentration-dependent response to IgG, thus eliminating the need for an enzyme-substrate system. Automated image analysis and quantification of antigen concentrations are able to be performed on Windows- and Android-based mobile platforms.

Aptamer-functionalized nanoparticles for surface immobilization-free electrochemical detection of cortisol in a microfluidic device.

Monitoring the periodic diurnal variations in cortisol from small volume samples of serum or saliva is of great interest, due to the regulatory role of cortisol within various physiological functions and stress symptoms. Current detection assays are immunologically based and require cumbersome antibody immobilization chemistries, thereby limiting the assay versatility, kinetics, and reproducibility. We present a quantitative aptamer-based detection methodology for cortisol that does not require target labeling, capture probe immobilization on the detection surface or wash steps prior to readout. Using a recognition system of aptamer functionalized gold nanoparticles pre-bound with electro-active triamcinolone, the cortisol level is detected based on its competitive binding to the aptamer by following signal from the displaced triamcinolone using square wave voltammetry at patterned graphene-modified electrodes in a microfluidic or nanoslit device. Due to the 3D analyte diffusion profile at the aptamer interface and the ability to enhance the surface area for cortisol capture, this assay shows signal linearity over a five-log analyte concentration range (10 μg/mL to 30 pg/mL) and exhibits rapid binding kinetics with cortisol versus other glucocorticoids, as apparent from the absence of interferences from estradiol, testosterone and progesterone. The assay is carried out within the biologically relevant range for glucocorticoids in serum and saliva matrices, and benchmarked versus ELISA and radioimmunoassays. Based on absence of cumbersome surface immobilization and wash steps for carrying out this assay, its quantitative signal characteristics and its ability to resist interferences from other glucocorticoids, we envision its application towards routine monitoring of cortisol within bio-fluids.

DNA Photonics [Deoxyribonucleic Acid]

ABSTRACT Purified deoxyribonucleic acid (DNA) derived from salmon and scallop sperm has demonstrated excellent passive and active optical properties. Characterization of the optical and electromagnetic properties of DNA suggests suitability for photonic applications. One of interesting features of DNA we discovered was an intercalation of aromatic compounds into stacked layers within the double helix of DNA molecules. We found that various optical dyes inserted into the double helix of DNA molecules rendered active optical waveguide materials with excellent nonlinear optical properties. Our research included the investigation of DNA for use as an optical waveguide material as well as intercalation of fluorescent dyes, photochromic dyes, nonlinear optic chromophores, two photon dyes and rare earth compounds into DNA for use as a nonlinear optical material.

Colorimetric detection with aptamer-gold nanoparticle conjugates coupled to an android-based color analysis application for use in the field

The feasibility of using aptamer-gold nanoparticle conjugates (Apt-AuNPs) to design colorimetric assays for in the field detection of small molecules was investigated. An assay to detect cocaine was designed using two clones of a known cocaine-binding aptamer. The assay was based on the AuNPs difference in affinity for single-stranded DNA (non-binding) and double stranded DNA (target bound). In the first assay, a commonly used design was followed, in which the aptamer and target were incubated to allow binding followed by exposure to the AuNPs. Interactions between the non-bound analytes and the AuNPs surface resulted in a number of false positives. The assay was redesigned by incubating the AuNPs and the aptamer prior to target addition to passivate the AuNPs surface. The adsorbed aptamer was able to bind the target while preventing non-specific interactions. The assay was validated with a number of masking and cutting agents and other controlled substances showing minimal false positives. Studies to improve the assay performance in the field were performed, showing that assay activity could be preserved for up to 2 months. To facilitate the assay analysis, an android application for automatic colorimetric characterization was developed. The application was validated by challenging the assay with cocaine standards of different concentrations, and comparing the results to a conventional plate reader, showing outstanding agreement. Finally, the rapid identification of cocaine in mixtures mimicking street samples was demonstrated. This work established that Apt-AuNPs can be used to design robust assays to be used in the field.

Paper pump for passive and programmable transport

In microfluidic systems, a pump for fluid-driving is often necessary. To keep the size of microfluidic systems small, a pump that is small in size, light-weight and needs no external power source is advantageous. In this work, we present a passive, simple, ultra-low-cost, and easily controlled pumping method based on capillary action of paper that pumps fluid through conventional polymer-based microfluidic channels with steady flow rate. By using inexpensive cutting tools, paper can be shaped and placed at the outlet port of a conventional microfluidic channel, providing a wide range of pumping rates. A theoretical model was developed to describe the pumping mechanism and aid in the design of paper pumps. As we show, paper pumps can provide steady flow rates from 0.3 μl/s to 1.7 μl/s and can be cascaded to achieve programmable flow-rate tuning during the pumping process. We also successfully demonstrate transport of the most common biofluids (urine, serum, and blood). With these capabilities, the paper pump has the potential to become a powerful fluid-driving approach that will benefit the fielding of microfluidic systems for point-of-care applications.

Biofunctionalized Zinc Oxide Field Effect Transistors for Selective Sensing of Riboflavin with Current Modulation

Zinc oxide field effect transistors (ZnO-FET), covalently functionalized with single stranded DNA aptamers, provide a highly selective platform for label-free small molecule sensing. The nanostructured surface morphology of ZnO provides high sensitivity and room temperature deposition allows for a wide array of substrate types. Herein we demonstrate the selective detection of riboflavin down to the pM level in aqueous solution using the negative electrical current response of the ZnO-FET by covalently attaching a riboflavin binding aptamer to the surface. The response of the biofunctionalized ZnO-FET was tuned by attaching a redox tag (ferrocene) to the 3′ terminus of the aptamer, resulting in positive current modulation upon exposure to riboflavin down to pM levels.

Pentacene organic thin-film transistors on flexible paper and glass substrates

Pentacene-based organic thin-film transistors (OTFTs) were fabricated on several types of flexible substrate: commercial photo paper, ultra-smooth specialty paper and ultra-thin (100 μM) flexible glass. The transistors were fabricated entirely through dry-step processing. The transconductance and field-effect mobility of OTFTs on photo paper reached values of ∼0.52 mS m(-1) and ∼ 0.1 cm(2) V (-1) s(-1), respectively. Preliminary results on the lifetime of OTFTs on photo paper yielded stable transconductance and mobility values over a period of more than 250 h. The comparable characteristics of OTFTs fabricated on widely available, low cost paper and high quality expensive liquid crystal display glass indicate the potential importance of cellulose-based electronic devices.