Wearable Flexible Perspiration Biosensors Using Laser-Induced Graphene and Polymeric Tape Microfluidics.

Wearable biosensors promise real-time measurements of chemicals in human sweat, with the potential for dramatic improvements in medical diagnostics and athletic performance through continuous metabolite and electrolyte monitoring. However, sweat sensing is still in its infancy, and questions remain about whether sweat can be used for medical purposes. Wearable sensors are focused on proof-of-concept designs that are not scalable for multisubject trials, which could elucidate the utility of sweat sensing for health monitoring. Moreover, many wearable sensors do not include the microfluidics necessary to protect and channel consistent and clean sweat volumes to the sensor surface or are not designed to be disposable to prevent sensor biofouling and inaccuracies due to repeated use. Hence, there is a need to produce low-cost and single-use wearable sensors with integrated microfluidics to ensure reliable sweat sensing. Herein, we demonstrate the convergence of laser-induced graphene (LIG) based sensors with soft tape polymeric microfluidics to quantify both sweat metabolites (glucose and lactate) and electrolytes (sodium) for potential hydration and fatigue monitoring. Distinct LIG-electrodes were functionalized with glucose oxidase and lactate oxidase for selective sensing of glucose and lactate across physiological ranges found in sweat with sensitivities of 26.2 and 2.47 × 10-3 µA mM-1 cm-2, detection limits of 8 and 220 µM, and linear response ranges of 0-1 mM and 0-32 mM, respectively. LIG-electrodes functionalized with a sodium-ion-selective membrane displayed Nernstian sensitivity of 58.8 mV decade-1 and a linear response over the physiological range in sweat (10-100 mM). The sensors were tested in a simulated sweating skin microfluidic system and on-body during cycling tests in a multisubject trial. Results demonstrate the utility of LIG integrated with microfluidics for real-time, continuous measurements of biological analytes in sweat and help pave the way for the development of personalized wearable diagnostic tools.

In situ digital holographic microscopy for rapid detection and monitoring of the harmful dinoflagellate, Karenia brevis.

Karenia brevis blooms, also known as red tide, are a recurring problem in the coastal Gulf of Mexico. These blooms have the capacity to inflict substantial damage to human and animal health as well as local economies. Thus, monitoring and detection of K. brevis blooms at all life stages and cell concentrations is essential for ensuring public safety. Current K. brevis monitoring methods have several limitations, including size resolution limits and concentration ranges, limited capacity for spatial and temporal profiling, and/or small sample volume processing. Here, a novel monitoring method wherein an autonomous digital holographic imaging microscope (AUTOHOLO), that overcomes these limitations and can characterize K. brevis concentrations in situ, is presented. Using the AUTOHOLO, in situ field measurements were conducted in the coastal Gulf of Mexico during an active K. brevis bloom over the 2020-21 winter season. Surface and sub-surface water samples collected during these field studies were also analyzed in the lab using benchtop holographic imaging and flow cytometry for validation. A convolutional neural network was trained for automated classification of K. brevis at all concentration ranges. The network was validated with manual counts and flow cytometry, yielding a 90% accuracy across diverse datasets with varying K. brevis concentrations. The usefulness of pairing the AUTOHOLO with a towing system was also demonstrated for characterizing particle abundance over large spatial distances, which could potentially facilitate characterization of spatial distributions of K. brevis during bloom events. Future applications of the AUTOHOLO can include integration into existing HAB monitoring networks to enhance detection capabilities for K. brevis in aquatic environments around the world.

In situ biological particle analyzer based on digital inline holography.

Obtaining in situ measurements of biological microparticles is crucial for both scientific research and numerous industrial applications (e.g., early detection of harmful algal blooms, monitoring yeast during fermentation). However, existing methods are limited to offer timely diagnostics of these particles with sufficient accuracy and information. Here, we introduce a novel method for real-time, in situ analysis using machine learning (ML)-assisted digital inline holography (DIH). Our ML model uses a customized YOLOv5 architecture specialized for the detection and classification of small biological particles. We demonstrate the effectiveness of our method in the analysis of 10 plankton species with equivalent high accuracy and significantly reduced processing time compared to previous methods. We also applied our method to differentiate yeast cells under four metabolic states and from two strains. Our results show that the proposed method can accurately detect and differentiate cellular and subcellular features related to metabolic states and strains. This study demonstrates the potential of ML-driven DIH approach as a sensitive and versatile diagnostic tool for real-time, in situ analysis of both biotic and abiotic particles. This method can be readily deployed in a distributive manner for scientific research and manufacturing on an industrial scale.

Tuning the Structure, Conductivity, and Wettability of Laser-Induced Graphene for Multiplexed Open Microfluidic Environmental Biosensing and Energy Storage Devices.

The integration of microfluidics and electrochemical cells is at the forefront of emerging sensors and energy systems; however, a fabrication scheme that can create both the microfluidics and electrochemical cells in a scalable fashion is still lacking. We present a one-step, mask-free process to create, pattern, and tune laser-induced graphene (LIG) with a ubiquitous CO2 laser. The laser parameters are adjusted to create LIG with different electrical conductivity, surface morphology, and surface wettability without the need for postchemical modification. Such definitive control over material properties enables the creation of LIG-based integrated open microfluidics and electrochemical sensors that are capable of dividing a single water sample along four multifurcating paths to three ion selective electrodes (ISEs) for potassium (K+), nitrate (NO3-), and ammonium (NH4+) monitoring and to an enzymatic pesticide sensor for organophosphate pesticide (parathion) monitoring. The ISEs displayed near-Nernstian sensitivities and low limits of detection (LODs) (10-5.01 M, 10-5.07 M, and 10-4.89 M for the K+, NO3-, and NH4+ ISEs, respectively) while the pesticide sensor exhibited the lowest LOD (15.4 pM) for an electrochemical parathion sensor to date. LIG was also specifically patterned and tuned to create a high-performance electrochemical micro supercapacitor (MSC) capable of improving the power density by 2 orders of magnitude compared to a Li-based thin-film battery and the energy density by 3 orders of magnitude compared to a commercial electrolytic capacitor. Hence, this tunable fabrication approach to LIG is expected to enable a wide range of real-time, point-of-use health and environmental sensors as well as energy storage/harvesting modules.

Laser-induced graphene electrodes for electrochemical ion sensing, pesticide monitoring, and water splitting.

Laser-induced graphene (LIG) has shown to be a scalable manufacturing route to create graphene electrodes that overcome the expense associated with conventional graphene electrode fabrication. Herein, we expand upon initial LIG reports by functionalizing the LIG with metallic nanoparticles for ion sensing, pesticide monitoring, and water splitting. The LIG electrodes were converted into ion-selective sensors by functionalization with poly(vinyl chloride)-based membranes containing K+ and H+ ionophores. These ion-selective sensors exhibited a rapid response time (10-15 s), near-Nernstian sensitivity (53.0 mV/dec for the K+ sensor and - 56.6 mV/pH for the pH sensor), and long storage stability for 40 days, and were capable of ion monitoring in artificial urine. The pesticide biosensors were created by functionalizing the LIG electrodes with the enzyme horseradish peroxidase and displayed a high sensitivity to atrazine (28.9 nA/µM) with negligible inference from other common herbicides (glyphosate, dicamba, and 2,4-dichlorophenoxyacetic acid). Finally, the LIG electrodes also exhibited a small overpotential for hydrogen evolution reaction and oxygen evolution reaction. The oxygen evolution reaction tests yielded overpotentials of 448 mV and 995 mV for 10 mA/cm2 and 100 mA/cm2, respectively. The hydrogen evolution reaction tests yielded 35 mV and 281 mV for the corresponding current densities. Such a versatile LIG platform paves the way for simple, efficient electrochemical sensing and energy harvesting applications.

Stamped multilayer graphene laminates for disposable in-field electrodes: application to electrochemical sensing of hydrogen peroxide and glucose.

A multi-step approach is described for the fabrication of multi-layer graphene-based electrodes without the need for ink binders or post-print annealing. Graphite and nanoplatelet graphene were chemically exfoliated using a modified Hummers' method and the dried material was thermally expanded. Expanded materials were used in a 3D printed mold and stamp to create laminate electrodes on various substrates. The laminates were examined for potential sensing applications using model systems of peroxide (H2O2) and enzymatic glucose detection. Within the context of these two assay systems, platinum nanoparticle electrodeposition and oxygen plasma treatment were examined as methods for improving sensitivity. Electrodes made from both materials displayed excellent H2O2 sensing capability compared to screen-printed carbon electrodes. Laminates made from expanded graphite and treated with platinum, detected H2O2 at a working potential of 0.3 V (vs. Ag/AgCl [0.1 M KCl]) with a 1.91 µM detection limit and sensitivity of 64 nA·µM-1·cm-2. Electrodes made from platinum treated nanoplatelet graphene had a H2O2 detection limit of 1.98 µM (at 0.3 V), and a sensitivity of 16.5 nA·µM-1·cm-2. Both types of laminate electrodes were also tested as glucose sensors via immobilization of the enzyme glucose oxidase. The expanded nanographene material exhibited a wide analytical range for glucose (3.7 µM to 9.9 mM) and a detection limit of 1.2 µM. The sensing range of laminates made from expanded graphite was slightly reduced (9.8 µM to 9.9 mM) and the detection limit for glucose was higher (18.5 µM). When tested on flexible substrates, the expanded graphite laminates demonstrated excellent adhesion and durability during testing. These properties make the electrodes adaptable to a variety of tests for field-based or wearable sensing applications. Graphical abstract Expanded graphite (eGR) and expanded nanoplatelet graphene (nGN) were chemically exfoliated, thermally expanded, and manually stamped into flexible multi-layer graphene laminate electrodes. Hydrogen peroxide amperometric testing of eGR laminates compared to nGN laminates and a screen printed carbon (SPC) electrode.