Continuous Flow with Reagent Injection on an Inlaid Microfluidic Platform Applied to Nitrite Determination.

A continuous flow with reagent injection method on a novel inlaid microfluidic platform for nitrite determination has been successfully developed. The significance of the high-frequency monitoring of nutrient fluctuations in marine environments is crucial for understanding our impacts on the ecosystem. Many in-situ systems face limitations in high-frequency data collection and have restricted deployment times due to high reagent consumption. The proposed microfluidic device employs automatic colorimetric absorbance spectrophotometry, using the Griess assay for nitrite determination, with minimal reagent usage. The sensor incorporates 10 solenoid valves, four syringes, two LEDs, four photodiodes, and an inlaid microfluidic technique to facilitate optical measurements of fluid volumes. In this flow system, Taylor-Aris dispersion was simulated for different injection volumes at a constant flow rate, and the results have been experimentally confirmed using red food dye injection into a carrier stream. A series of tests were conducted to determine a suitable injection frequency for the reagent. Following the initial system characterization, seven different standard concentrations ranging from 0.125 to 10 µM nitrite were run through the microfluidic device to acquire a calibration curve. Three different calibrations were performed to optimize plug length, with reagent injection volumes of 4, 20, and 50 µL. A straightforward signal processing method was implemented to mitigate the Schlieren effect caused by differences in refractive indexes between the reagent and standards. The results demonstrate that a sampling frequency of at least 10 samples per hour is achievable using this system. The obtained attenuation coefficients exhibited good agreement with the literature, while the reagent consumption was significantly reduced. The limit of detection for a 20 µL injection volume was determined to be 94 nM from the sample intake, and the limit of quantification was 312 nM. Going forward, the demonstrated system will be packaged in a submersible enclosure to facilitate in-situ colorimetric measurements in marine environments.

Compact and automated eDNA sampler for in situ monitoring of marine environments.

Using environmental DNA (eDNA) to monitor biodiversity in aquatic environments is becoming an efficient and cost-effective alternative to other methods such as visual and acoustic identification. Until recently, eDNA sampling was accomplished primarily through manual sampling methods; however, with technological advances, automated samplers are being developed to make sampling easier and more accessible. This paper describes a new eDNA sampler capable of self-cleaning and multi-sample capture and preservation, all within a single unit capable of being deployed by a single person. The first in-field test of this sampler took place in the Bedford Basin, Nova Scotia, Canada alongside parallel samples taken using the typical Niskin bottle collection and post-collection filtration method. Both methods were able to capture the same aquatic microbial community and counts of representative DNA sequences were well correlated between methods with R[Formula: see text] values ranging from 0.71-0.93. The two collection methods returned the same top 10 families in near identical relative abundance, demonstrating that the sampler was able to capture the same community composition of common microbes as the Niskin. The presented eDNA sampler provides a robust alternative to manual sampling methods, is amenable to autonomous vehicle payload constraints, and will facilitate persistent monitoring of remote and inaccessible sites.

An Automated Microfluidic Analyzer for In Situ Monitoring of Total Alkalinity.

We have designed, built, tested, and deployed an autonomous in situ analyzer for seawater total alkalinity. Such analyzers are required to understand the ocean carbon cycle, including anthropogenic carbon dioxide (CO2) uptake and for mitigation efforts via monitoring, reporting, and verification of carbon dioxide removal through ocean alkalinity enhancement. The microfluidic nature of our instrument makes it relatively lightweight, reagent efficient, and amenable for use on platforms that would carry it on long-term deployments. Our analyzer performs a series of onboard closed-cell titrations with three independent stepper-motor driven syringe pumps, providing highly accurate mixing ratios that can be systematically swept through a range of pH values. Temperature effects are characterized over the range 5-25 °C allowing for field use in most ocean environments. Each titration point requires approximately 170 µL of titrant, 830 µL of sample, 460 J of energy, and a total of 105 s for pumping and optical measurement. The analyzer performance is demonstrated through field data acquired at two sites, representing a cumulative 25 days of operation, and is evaluated against laboratory measurements of discrete water samples. Once calibrated against onboard certified reference material, the analyzer showed an accuracy of -0.17 ± 24 µmol kg-1. We further report a precision of 16 µmol kg-1, evaluated on repeated in situ measurements of the aforementioned certified reference material. The total alkalinity analyzer presented here will allow measurements to take place in remote areas over extended periods of time, facilitating affordable observations of a key parameter of the ocean carbon system with high spatial and temporal resolution.

A Magnetically Tunable Check Valve Applied to a Lab-on-Chip Nitrite Sensor.

Presented here is the fabrication and characterization of a tunable microfluidic check valve for use in marine nutrient sensing. The ball-style valve makes use of a rare-earth permanent magnet, which exerts a pulling force to ensure it remains passively sealed until the prescribed cracking pressure is met. By adjusting the position of the magnet, the cracking pressure is shown to be customizable to meet design requirements. Further applicability is shown by integrating the valve into a poly(methyl methacrylate) (PMMA) lab-on-chip device with an integrated optical absorbance cell for nitrite detection in seawater. Micro-milling is used to manufacture both the valve and the micro-channel structures. The valve is characterized up to a flow rate of 14 mL min-1 and exhibits low leakage rates at high back pressures (<2 µL min-1 at ~350 kPa). It is low cost, requires no power, and is easily implemented on microfluidic platforms.