PCL/PEO Polymer Membrane Prevents Biofouling in Wearable Detection Sensors.

Technological advances in biosensing offer extraordinary opportunities to transfer technologies from a laboratory setting to clinical point-of-care applications. Recent developments in the field have focused on electrochemical and optical biosensing platforms. Unfortunately, these platforms offer relatively poor sensitivity for most of the clinically relevant targets that can be measured on the skin. In addition, the non-specific adsorption of biomolecules (biofouling) has proven to be a limiting factor compromising the longevity and performance of these detection systems. Research from our laboratory seeks to capitalize on analyte selective properties of biomaterials to achieve enhanced analyte adsorption, enrichment, and detection. Our goal is to develop a functional membrane integrated into a microfluidic sampling interface and an electrochemical sensing unit. The membrane was manufactured from a blend of Polycaprolactone (PCL) and Polyethylene oxide (PEO) through a solvent casting evaporation method. A microfluidic flow cell was developed with a micropore array that allows liquid to exit from all pores simultaneously, thereby imitating human perspiration. The electrochemical sensing unit consisted of planar gold electrodes for the monitoring of nonspecific adsorption of proteins utilizing Cyclic Voltammetry (CV) and Electrochemical Impedance Spectroscopy (EIS). The solvent casting evaporation technique proved to be an effective method to produce membranes with the desired physical properties (surface properties and wettability profile) and a highly porous and interconnected structure. Permeability data from the membrane sandwiched in the flow cell showed excellent permeation and media transfer efficiency with uniform pore activation for both active and passive sweat rates. Biofouling experiments exhibited a decrease in the extent of biofouling of electrodes protected with the PCL/PEO membrane, corroborating the capacity of our material to mitigate the effects of biofouling.

Fast Adhesion of Gold Nanoparticles (AuNPs) to a Surface Using Starch Hydrogels for Characterization of Biomolecules in Biosensor Applications.

Gold nanoparticles (AuNPs) are the most thoroughly studied nanoparticles because of their remarkable optical properties. Color changes in assays that use AuNPs can be easily observed with the naked eye, resulting in sensitive colorimetric methods, useful for detecting a variety of biological molecules. However, while AuNPs represent an excellent nano-platform for developing analytical methods for biosensing, there are still challenges that must be overcome before colloidal AuNPs formulation can be successfully translated into practical applications. One of those challenges is the ability to immobilize AuNPs in a solid support. There are many difficulties with controlling both the cluster size and the adhesion of the coatings formed. In addition, many of the techniques employed are expensive and time-consuming, or require special equipment. Thus, a simple and inexpensive method that only requires common lab equipment for immobilizing AuNPs on a surface using Starch Hydrogels has been developed. Starch hydrogels confer a 400% increase in stability to the nanoparticles when exposed to changes in the environment while also allowing for macromolecules to interact with the AuNPs surface. Several starch derivatives were tested, including, dextrin, beta-cyclodextrin and maltodextrin, being dextrin the one that conferred the highest stability. As a proof-of-concept, a SlipChip microfluidic sensor scheme was developed to measure the concentration of DNA in a sample. The detection limit of our biosensor was found to be 25 ng/mL and 75 ng/mL for instrument and naked eye detection, respectively.

Flow characterization and patch clamp dose responses using jet microfluidics in a tubeless microfluidic device.

BACKGROUND: Surface tension passive pumping is a way to actuate flow without the need for pumps, tubing or valves by using the pressure inside small drop to move liquid via a microfluidic channel. These types of tubeless devices have typically been used in cell biology. Herein we present the use of tubeless devices as a fluid exchange platform for patch clamp electrophysiology. NEW METHOD: Inertia from high-speed droplets and jets is used to create flow and perform on-the-fly mixing of solutions. These are then flowed over GABA transfected HEK cells under patch in order to perform a dose response analysis. RESULTS: TIRF imaging and electrical recordings are used to study the fluid exchange properties of the microfluidic device, resulting in 0-90% fluid exchange times of hundreds of milliseconds. COMSOL is used to model flow and fluid exchange within the device. Patch-clamping experiments show the ability to use high-speed passive pumping and its derivatives for studying peak dose responses, but not for studying ion channel kinetics. COMPARISON WITH EXISTING METHOD(S): Our system results in fluid exchange times slower than when using a standard 12-barrel application system and is not as stable as traditional methods, but it offers a new platform with added functionality. CONCLUSIONS: Surface tension passive pumping and tubeless devices can be used in a limited fashion for electrophysiology. Users may obtain peak dose responses but the system, in its current form, is not capable of fluid exchange fast enough to study the kinetics of most ion channels.

Detection of ESKAPE Bacterial Pathogens at the Point of Care Using Isothermal DNA-Based Assays in a Portable Degas-Actuated Microfluidic Diagnostic Assay Platform.

An estimated 1.5 billion microbial infections occur globally each year and result in ∼4.6 million deaths. A technology gap associated with commercially available diagnostic tests in remote and underdeveloped regions prevents timely pathogen identification for effective antibiotic chemotherapies for infected patients. The result is a trial-and-error approach that is limited in effectiveness, increases risk for patients while contributing to antimicrobial drug resistance, and reduces the lifetime of antibiotics. This paper addresses this important diagnostic technology gap by describing a low-cost, portable, rapid, and easy-to-use microfluidic cartridge-based system for detecting the ESKAPE (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter spp.) bacterial pathogens that are most commonly associated with antibiotic resistance. The point-of-care molecular diagnostic system consists of a vacuum-degassed microfluidic cartridge preloaded with lyophilized recombinase polymerase amplification (RPA) assays and a small portable battery-powered electronic incubator/reader. The isothermal RPA assays detect the targeted ESKAPE pathogens with high sensitivity (e.g., a limit of detection of ∼10 nucleic acid molecules) that is comparable to that of current PCR-based assays, and they offer advantages in power consumption, engineering, and robustness, which are three critical elements required for the point-of-care setting. IMPORTANCE: This paper describes a portable system for rapidly identifying bacteria in resource-limited environments; we highlight the capabilities of the technology by detecting different pathogens within the ESKAPE collection, which cause nosocomial infections. The system is designed around isothermal DNA-based assays housed within an autonomous plastic cartridge that are designed with the end user in mind, who may have limited technological training. Displaying excellent sensitivity and specificity, the assay systems that we demonstrate may enable future diagnoses of bacterial infection to guide the development of effective chemotherapies and may have a role in areas beyond health where rapid detection is valuable, including in industrial processing and manufacturing, food security, agriculture, and water quality testing.

An inertia enhanced passive pumping mechanism for fluid flow in microfluidic devices.

We describe and characterize a pumping mechanism that leverages the momentum present in small droplets ejected from a micro-nozzle to drive flow in an open microfluidic device. This approach allows driving flow in a microfluidic device in a regime that offers unique features different to those achievable with typical passive pumping or syringe-pump driven flow. Two flow regimes with specific flow characteristics are described: inertia enhanced passive pumping, in which fluid exchange times in the channel are significantly reduced, and inertia actuated flow, in which it is possible to initiate flow in an empty channel or against natural pressure gradients. Momentum is leveraged to create rapid fluid exchanges, instantaneous flow reversal, filling and mixing inside the microfluidic device.

An automated microdroplet passive pumping platform for high-speed and packeted microfluidic flow applications.

Surface tension driven passive pumping is a microfluidic technology that uses the surface tension present in small droplets to generate flow. To enhance the potential of this type of passive pumping, a new 'micro passive pumping' technique has been developed that allows for high throughput fluidic delivery by combining passive pumping with a small droplet-based fluidic ejection system. Flow rates of up to four milliliters per minute (mL/min) were achieved that are solely limited by the channel geometry and droplet size. Fluid exchange rates can be performed within tens of milliseconds (ms) by delivering fluids from multiple nozzles. The technique can be extended to a multitude of platforms, as channels are not pressurized and therefore do not require bonding to a substrate. This technique provides a novel flow control for high-speed and packeted flow applications without requiring external tubing connections or substrate bonding.

High speed droplet-based delivery system for passive pumping in microfluidic devices.

A novel microfluidic system has been developed that uses the phenomenon of passive pumping along with a user controlled droplet based fluid delivery system. Passive pumping is the phenomenon by which surface tension induced pressure differences drive fluid movement in closed channels. The automated fluid delivery system consists of a set of voltage controlled valves with micro-nozzles connected to a fluid reservoir and a control system. These voltage controlled valves offer a volumetrically precise way to deliver fluid droplets to the inlet of a microfluidic device in a high frequency manner. Based on the dimensions demonstrated in the current study example, the system is capable of flowing 4 milliliters per minute (through a 2.2 mm by 260 microm cross-sectional channel). Based on these same channel dimensions, fluid exchange of a point inside the channel can be achieved in as little as eight milliseconds. It is observed that there is interplay between momentum of the system (imparted by a combination of the droplets created by the valves and the fluid velocity in the channel), and the surface tension of the liquid. Where momentum provides velocity to the fluid flow (or vice-versa), equilibration of surface tension at the inlet provides a sudden stop to any flow. This sudden stop allows the user to control the flow characteristics of the channel and opens the door for a variety of biological applications, ranging anywhere from reagent delivery to drug-cell studies. It is also observed that when nozzles are aimed at the inlet at shallow angles, the droplet momentum can cause additional interesting fluid phenomena, such as mixing of multiple droplets in the inlet.