Biosignature Molecules Accumulate and Persist in Evaporitic Brines: Implications for Planetary Exploration.

The abundance of potentially habitable hypersaline environments in our solar system compels us to understand the impacts of high-salt matrices and brine dynamics on biosignature detection efforts. We identified and quantified organic compounds in brines from South Bay Salt Works (SBSW), where evapoconcentration of ocean water enables exploration of the impact of NaCl- and MgCl-dominated brines on the detection of potential biosignature molecules. In SBSW, organic biosignature abundance and distribution are likely influenced by evapoconcentration, osmolyte accumulation, and preservation effects.

A Robust Capillary Electrophoresis with Laser-Induced Fluorescence Detection (CE-LIF) Method for Quantitative Compositional Analysis of Trace Amino Acids in Hypersaline Samples.

The search for life in our solar system can be enabled by the characterization of extreme environments representing conditions expected on other planets within our solar system. Molecular abundances observed in these environments help establish instrument design requirements, including limits of detection and pH/salt tolerance, and may be used for validation of proposed planetary science instrumentation. Here, we optimize capillary electrophoresis with laser-induced fluorescence detection (CE-LIF) separations for low limit of detection quantitative compositional analysis of amino acids in hypersaline samples using carboxyfluorescein succinimidyl ester (CFSE) as the amine-reactive fluorescent probe.

Analytical Chemistry Throughout This Solar System.

One of the greatest and most long-lived scientific pursuits of humankind has been to discover and study the planetary objects comprising our solar system. Information gained from solar system observations, via both remote sensing and in situ measurements, is inherently constrained by the analytical (often chemical) techniques we employ in these endeavors. The past 50 years of planetary science missions have resulted in immense discoveries within and beyond our solar system, enabled by state-of-the-art analytical chemical instrument suites on board these missions.

A miniaturized, low-cost lens tube based laser-induced fluorescence detection system for automated microfluidic analysis of primary amines.

In situ analyses are essential to ascertain potential past or present habitability in celestial bodies. One technique that provides the sensitivity and miniaturization needed to successfully detect trace organics in the outer Solar System is laser-induced fluorescence (LIF) detection, which, when coupled with microfluidic systems, provides a powerful wet chemistry platform that can meet the size and resource consumption constraints of a remote analysis mission. Herein, a portable LIF detection module (44-mm long, 18-mm wide) was prototyped and utilized to quantify bulk organics in a liquid sample via manual and automated analysis utilizing a programmable microfluidic architecture.

Analytical Chemistry in Astrobiology.

This Feature introduces and discusses the findings of key analytical techniques used to study planetary bodies in our solar system in the search for life beyond Earth, future missions planned for high-priority astrobiology targets in our solar system, and the challenges we face in performing these investigations.

A modular, easy-to-use microcapillary electrophoresis system with laser-induced fluorescence for quantitative compositional analysis of trace organic molecules.

Microcapillary electrophoresis (μCE) enables high-resolution separations in miniaturized, automated microfluidic devices. Pairing this powerful separation technique with laser-induced fluorescence (LIF) enables a highly sensitive, quantitative, and compositional analysis of organic molecule monomers and short polymers, which are essential, ubiquitous components of life on Earth. Improving methods for their detection has applications to multiple scientific fields, particularly those related to medicine, industry, and space science.

A practical guide to rapid-prototyping of PDMS-based microfluidic devices: A tutorial.

Micro total analytical systems (μTAS) are attractive to multiple fields that include chemistry, medicine and engineering due to their portability, low power usage, potential for automation, and low sample and reagent consumption, which in turn results in low waste generation. The development of fully-functional μTAS is an iterative process, based on the design, fabrication and testing of multiple prototype microdevices. Typically, microfabrication protocols require a week or more of highly-skilled personnel time in high-maintenance cleanroom facilities, which makes this iterative process cost-prohibitive in many locations worldwide.

Feasibility of Detecting Bioorganic Compounds in Enceladus Plumes with the Enceladus Organic Analyzer.

Enceladus presents an excellent opportunity to detect organic molecules that are relevant for habitability as well as bioorganic molecules that provide evidence for extraterrestrial life because Enceladus' plume is composed of material from the subsurface ocean that has a high habitability potential and significant organic content. A primary challenge is to send instruments to Enceladus that can efficiently sample organic molecules in the plume and analyze for the most relevant molecules with the necessary detection limits. To this end, we present the scientific feasibility and engineering design of the Enceladus Organic Analyzer (EOA) that uses a microfluidic capillary electrophoresis system to provide sensitive detection of a wide range of relevant organic molecules, including amines, amino acids, and carboxylic acids, with ppm plume-detection limits (100 pM limits of detection).

Technical aspects and challenges of colorimetric detection with microfluidic paper-based analytical devices (μPADs) - A review.

Paper-based devices are a leading alternative among the main analytical tools for point-of-care testing, due to their portability, low-cost, and ease-of-use. Colorimetric readouts are the most common method of detection in these microfluidic devices, enabling qualitative, semi-quantitative and fully quantitative analysis of multiple analytes. There is a multitude of ways to obtain a colorimetric output in such devices, including nanoparticles, dyes, redox and pH indicators, and each has unique drawbacks and benefits.

Improving Sample Distribution Homogeneity in Three-Dimensional Microfluidic Paper-Based Analytical Devices by Rational Device Design.

Paper-based devices are a portable, user-friendly, and affordable technology that is one of the best analytical tools for inexpensive diagnostic devices. Three-dimensional microfluidic paper-based analytical devices (3D-μPADs) are an evolution of single layer devices and they permit effective sample dispersion, individual layer treatment, and multiplex analytical assays. Here, we present the rational design of a wax-printed 3D-μPAD that enables more homogeneous permeation of fluids along the cellulose matrix than other existing designs in the literature.

Pneumatically actuated microvalve circuits for programmable automation of chemical and biochemical analysis.

Programmable microfluidic platforms (PMPs) are enabling significant advances in the utility of microfluidics for chemical and biochemical analysis. Traditional microfluidic devices are analogous to application-specific devices--a new device is needed to implement each new chemical or biochemical assay. PMPs are analogous to digital electronic processors--all that is needed to implement a new assay is a change in the order of operations conducted by the device.

Amine Analysis Using AlexaFluor 488 Succinimidyl Ester and Capillary Electrophoresis with Laser-Induced Fluorescence.

Fluorescent probes enable detection of otherwise nonfluorescent species via highly sensitive laser-induced fluorescence. Organic amines are predominantly nonfluorescent and are of analytical interest in agricultural and food science, biomedical applications, and biowarfare detection. Alexa Fluor 488 N-hydroxysuccinimidyl ester (AF488 NHS-ester) is an amine-specific fluorescent probe.

Analysis of thiols by microchip capillary electrophoresis for in situ planetary investigations.

Microchip capillary electrophoresis with laser-induced fluorescence detection (μCE-LIF) enables sensitive analyses of a wide range of analytes employing small volumes of sample and reagent (nL to μL) on an instrument platform with minimal mass, volume, and power requirements. This technique has been used previously in the analysis of amino acids and other organic molecules of interest in the fields of astrobiology and planetary science. Here, we present a protocol for the analysis of thiols using μCE-LIF.

Universal microfluidic automaton for autonomous sample processing: application to the Mars Organic Analyzer.

A fully integrated multilayer microfluidic chemical analyzer for automated sample processing and labeling, as well as analysis using capillary zone electrophoresis is developed and characterized. Using lifting gate microfluidic control valve technology, a microfluidic automaton consisting of a two-dimensional microvalve cellular array is fabricated with soft lithography in a format that enables facile integration with a microfluidic capillary electrophoresis device. The programmable sample processor performs precise mixing, metering, and routing operations that can be combined to achieve automation of complex and diverse assay protocols.

Low-temperature microchip nonaqueous capillary electrophoresis of aliphatic primary amines: applications to Titan chemistry.

We demonstrate microchip nonaqueous capillary electrophoresis (μNACE) analysis of primary aliphatic amines (C1-C18) in ethanol down to -20 °C as a first step in adapting microfluidic protocols for in situ analysis on Titan. To our knowledge, this is the first report of a nonaqueous separation at -20 °C on-chip. Limits of detection (LODs) ranged from 1.

Digitally programmable microfluidic automaton for multiscale combinatorial mixing and sample processing.

A digitally programmable microfluidic Automaton consisting of a 2-dimensional array of pneumatically actuated microvalves is programmed to perform new multiscale mixing and sample processing operations. Large (μL-scale) volume processing operations are enabled by precise metering of multiple reagents within individual nL-scale valves followed by serial repetitive transfer to programmed locations in the array. A novel process exploiting new combining valve concepts is developed for continuous rapid and complete mixing of reagents in less than 800 ms.

Analysis of thiols by microchip capillary electrophoresis for in situ planetary investigations.

The detection of thiols on extraterrestrial bodies could provide evidence for life, as well as a host of potential prebiological or abiological processes. Here, we report a novel protocol to analyze organic thiols by microchip CE with LIF detection. Thiols were labeled with Pacific Blue C5 maleimide and analyzed by MEKC.

Microchip capillary electrophoresis instrumentation for in situ analysis in the search for extraterrestrial life.

The search for signs of life on extraterrestrial planetary bodies is among NASA's top priorities in Solar System exploration. The associated pursuit of organics and biomolecules as evidence of past or present life demands in situ investigations of planetary bodies for which sample return missions are neither practical nor affordable. These in situ studies require instrumentation capable of sensitive chemical analyses of complex mixtures including a broad range of organic molecules.

Toward total automation of microfluidics for extraterrestial in situ analysis.

Despite multiple orbiter and landed missions to extraterrestrial bodies in the solar system, including Mars and Titan, we still know relatively little about the detailed chemical composition and quantity of organics and biomolecules in those bodies. For chemical analysis on astrobiologically relevant targets such as Mars, Europa, Titan, and Enceladus, instrumentation should be extremely sensitive and capable of analyzing a broad range of organic molecules. Microchip capillary electrophoresis (μCE) with laser-induced fluorescence (LIF) detection provides this required sensitivity and targets a wide range of relevant markers but, to date, has lacked the necessary degree of automation for spaceflight applications.

Analysis of carbonaceous biomarkers with the Mars Organic Analyzer microchip capillary electrophoresis system: carboxylic acids.

The oxidizing surface chemistry on Mars argues that any comprehensive search for organic compounds indicative of life requires methods to analyze higher oxidation states of carbon with very low limits of detection. To address this goal, microchip capillary electrophoresis (μCE) methods were developed for analysis of carboxylic acids with the Mars Organic Analyzer (MOA). Fluorescent derivatization was achieved by activation with the water soluble 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) followed by reaction with Cascade Blue hydrazide in 30 mM borate, pH 3.

Analysis of carbonaceous biomarkers with the Mars Organic Analyzer microchip capillary electrophoresis system: aldehydes and ketones.

A microchip CE method is developed for the analysis of two oxidized forms of carbon, aldehydes and ketones, with the Mars Organic Analyzer (MOA). Fluorescent derivitization is achieved in ∼ 15 min by hydrazone formation with Cascade Blue hydrazide in 30 mM borate pH 5-6. The microchip CE separation and analysis method is optimized via separation in 30 mM borate buffer, pH 9.

Multichannel capillary electrophoresis microdevice and instrumentation for in situ planetary analysis of organic molecules and biomarkers.

The Multichannel Mars Organic Analyzer (McMOA), a portable instrument for the sensitive microchip capillary electrophoresis (CE) analysis of organic compounds such as amino acid biomarkers and polycyclic aromatic hydrocarbons (PAHs), is developed. The instrument uses a four-layer microchip, containing eight CE analysis systems integrated with a microfluidic network for autonomous fluidic processing. The McMOA has improved optical components that integrate 405 nm laser excitation with a linear-scanning optical system capable of multichannel real-time fluorescence spectroscopic analysis.

Capillary electrophoresis analysis of organic amines and amino acids in saline and acidic samples using the Mars organic analyzer.

The Mars Organic Analyzer (MOA) has enabled the sensitive detection of amino acid and amine biomarkers in laboratory standards and in a variety of field sample tests. However, the MOA is challenged when samples are extremely acidic and saline or contain polyvalent cations. Here, we have optimized the MOA analysis, sample labeling, and sample dilution buffers to handle such challenging samples more robustly.

Enhanced amine and amino acid analysis using Pacific Blue and the Mars Organic Analyzer microchip capillary electrophoresis system.

The fluorescent amine reactive probe Pacific Blue succinimidyl ester (PB) is used for the detection of trace amounts of amines and amino acids by microchip capillary electrophoresis on the Mars Organic Analyzer (MOA). The spectral and chemical properties of PB provide a 200-fold increase in sensitivity and improved resolution compared to fluorescamine derivatization. With the use of cross injection and PB labeling, the MOA detected amino acids at concentrations as low as 75 pM (sub-parts-per-trillion).

Polycyclic aromatic hydrocarbon analysis with the Mars organic analyzer microchip capillary electrophoresis system.

The Mars Organic Analyzer (MOA), a portable microchip capillary electrophoresis (CE) instrument developed for sensitive amino acid analysis on Mars, is used to analyze laboratory standards and real-world samples for polycyclic aromatic hydrocarbons (PAHs). The microfabricated CE separation and analysis method for these hydrophobic analytes is optimized, resulting in a separation buffer consisting of 10 mM sulfobutylether-beta-cyclodextrin, 40 mM methyl-beta-cyclodextrin, 5 mM carbonate buffer at pH 10, 5 degrees C. A PAH standard consisting of seven PAHs found in extraterrestrial matter and two terrestrial PAHs is successfully baseline separated.