Integrated high performance microfluidic organic analysis instrument for planetary and space exploration.

The exploration of our solar system to characterize the molecular organic inventory will enable the identification of potentially habitable regions and initiate the search for biosignatures of extraterrestrial life. However, it is challenging to perform the required high-resolution, high-sensitivity chemical analyses in space and in planetary environments. To address this challenge, we have developed a microfluidic organic analyzer (MOA) instrument that consists of a multilayer programmable microfluidic analyzer (PMA) for fluidic processing at the microliter scale coupled with a microfabricated glass capillary electrophoresis (CE) wafer for separation and analysis of the sample components. Organic analytes are labeled with a functional group-specific (e.g. amine, organic acid, aldehyde) fluorescent dye, separated according to charge and hydrodynamic size by capillary electrophoresis (CE), and detected with picomolar limit of detection (LOD) using laser-induced fluorescence (LIF). Our goal is a sensitive automated instrument and autonomous process that enables sample-in to data-out performance in a flight capable format. We present here the design, fabrication, and operation of a technology development unit (TDU) that meets these design goals with a core mass of 3 kg and a volume of <5 L. MOA has a demonstrated resolution of 2 × 105 theoretical plates for relevant amino acids using a 15 cm long CE channel and 467 V cm-1. The LOD of LIF surpasses 100 pM (0.01 ppb), enabling biosignature detection in harsh environments on Earth. MOA is ideally suited for probing biosignatures in potentially habitable destinations on icy moons such as Europa and Enceladus, and on Mars.

Operation of a programmable microfluidic organic analyzer under microgravity conditions simulating space flight environments.

A programmable microfluidic organic analyzer was developed for detecting life signatures beyond Earth and clinical monitoring of astronaut health. Extensive environmental tests, including various gravitational environments, are required to confirm the functionality of this analyzer and advance its overall Technology Readiness Level. This work examines how the programmable microfluidic analyzer performed under simulated Lunar, Martian, zero, and hypergravity conditions during a parabolic flight. We confirmed that the functionality of the programmable microfluidic analyzer was minimally affected by the significant changes in the gravitational field, thus paving the way for its use in a variety of space mission opportunities.

Optimization of Fluorescence Labeling of Trace Analytes: Application to Amino Acid Biosignature Detection with Pacific Blue.

Fluorescence labeling of biomolecules and fluorescence detection platforms provide a powerful approach to high-sensitivity bioanalysis. Reactive probes can be chosen to target specific functional groups to enable selective analysis of a chosen class of analytes. Particularly, when targeting trace levels of analytes, it is important to optimize the reaction chemistry to maximize the labeling efficiency and minimize the background. Here, we develop methods to optimize the labeling and detection of Pacific Blue (PB)-tagged amino acids. A model is developed to quantitate labeling kinetics and completeness in the circumstance where analyte labeling and reactive probe hydrolysis are in competition. The rates of PB hydrolysis and amino acid labeling are determined as a function of pH. Labeling kinetics and completeness as a function of PB concentration are found to depend on the ratio of the hydrolysis time to the initial labeling time, which depends on the initial PB concentration. Finally, the optimized labeling and detection conditions are used to perform capillary electrophoresis analysis demonstrating 100 pM sensitivities and high-efficiency separations of an 11 amino acid test set. This work provides a quantitative optimization model that is applicable to a variety of reactive probes and targets. Our approach is particularly useful for the analysis of trace amine and amino acid biosignatures in extraterrestrial samples. For illustration, our optimized conditions (reaction at 4 °C in a pH 8.5 buffer) are used to detect trace amino acid analytes at the 100 pM level in an Antarctic ice core sample.

Quantitative evaluation of the feasibility of sampling the ice plumes at Enceladus for biomarkers of extraterrestrial life.

Enceladus, an icy moon of Saturn, is a compelling destination for a probe seeking biosignatures of extraterrestrial life because its subsurface ocean exhibits significant organic chemistry that is directly accessible by sampling cryovolcanic plumes. State-of-the-art organic chemical analysis instruments can perform valuable science measurements at Enceladus provided they receive sufficient plume material in a fly-by or orbiter plume transit. To explore the feasibility of plume sampling, we performed light gas gun experiments impacting micrometer-sized ice particles containing a fluorescent dye biosignature simulant into a variety of soft metal capture surfaces at velocities from 800 m ⋅ s-1 up to 3 km ⋅ s-1 Quantitative fluorescence microscopy of the capture surfaces demonstrates organic capture efficiencies of up to 80 to 90% for isolated impact craters and of at least 17% on average on indium and aluminum capture surfaces at velocities up to 2.2 km ⋅ s-1 Our results reveal the relationships between impact velocity, particle size, capture surface, and capture efficiency for a variety of possible plume transit scenarios. Combined with sensitive microfluidic chemical analysis instruments, we predict that our capture system can be used to detect organic molecules in Enceladus plume ice at the 1 nM level-a sensitivity thought to be meaningful and informative for probing habitability and biosignatures.

Method for detecting and quantitating capture of organic molecules in hypervelocity impacts.

Enceladus is a prime candidate in the solar system for in-depth astrobiological studies searching for habitability and life because it has a liquid water ocean with significant organic content and ongoing cryovolcanic activity. The presence of ice plumes that jet up through fissures in the ice crust covering the sub-surface ocean, enables remote sampling and in-situ analysis via a fly-by mission. However, capture and transport of organic materials to organic analyzers presents distinctive challenges as it is unknown whether, and to what extent, organic molecules imbedded in ice particles can be captured and survive hypervelocity impacts. This manuscript provides a fluorescence microscopic method to parametrically determine the amount of an organic fluorescent tracer dye, Pacific Blue™ (PB) deposited on a metallic surface. This method can be used to measure the capture and survival outcomes of terrestrial hypervelocity impact experiments where an ice projectile labeled with Pacific Blue impacts a soft metal surface. This work is an important step in the advancement of instruments like the Enceladus Organic Analyzer for detecting biosignatures in an Enceladus plume fly-by mission. An apparatus consisting of a substrate humidification shroud coupled with an epifluorescence microscope with CCD detector is developed to measure the intensity of quantitatively deposited Pacific Blue droplets under controlled humidity. Calibration curves are produced that relate the integrated fluorescence intensity of humidified PB droplets on metal foils to the number of PB molecules deposited. To demonstrate the utility of this method, our calibrations are used to analyze and quantitate organic capture and survival (up to 11% capture efficiency) following ice particle impacts at a velocity of 1.7 km/s on an aluminum substrate.

Fabrication of high-quality glass microfluidic devices for bioanalytical and space flight applications.

Microfabricated glass microfluidic and Capillary Electrophoresis (CE) devices have been utilized in a wide variety of applications over the past thirty years. At the Berkeley Space Sciences Laboratory, we are working to further expand this technology by developing analytical instruments to chemically explore our solar system. This effort requires improving the quality and reliability of glass microfabrication through quality control procedures at every stage of design and manufacture. This manuscript provides detailed information on microfabrication technology for the production of high-quality glass microfluidic chips in compliance with industrial standards and space flight instrumentation quality control.•The methodological protocol provided in this paper includes the scope of each step of the manufacturing process, materials and technologies recommended and the specific challenges that often confront the process developer.•Types and sources of fabrication error at every stage have been identified and their solutions have been proposed and verified.•We present robust and rigorous manufacturing and quality control procedures that will assist other researchers in achieving the highest possible quality glass microdevices using the latest apparatus in a routine and reliable fashion.

Constraints on the formation environment of two chondrule-like igneous particles from Comet 81P/Wild 2.

Using chemical and petrologic evidence and modeling, we deduce that two chondrule-like particles named Iris and Callie, from Stardust cometary track C2052,12,74, formed in an environment very similar to that seen for type II chondrules in meteorites. Iris was heated near liquidus, equilibrated, and cooled at ≤ 100 °C/hr and within ≈ 2 log units of the IW buffer with a high partial pressure of Na such as would be present with dust enrichments of ≈ 103. There was no detectable metamorphic, nebular or aqueous alteration. In previous work Ogliore et al. (2012) reported that Iris formed late, > 3 Myr after CAIs, assuming 26Al was homogenously distributed, and was rich in heavy oxygen. Iris may be similar to assemblages found only in interplanetary dust particles and Stardust cometary samples called Kool particles. Callie is chemically and isotopically very similar but not identical to Iris.

Interstellar dust. Evidence for interstellar origin of seven dust particles collected by the Stardust spacecraft.

Seven particles captured by the Stardust Interstellar Dust Collector and returned to Earth for laboratory analysis have features consistent with an origin in the contemporary interstellar dust stream. More than 50 spacecraft debris particles were also identified. The interstellar dust candidates are readily distinguished from debris impacts on the basis of elemental composition and/or impact trajectory. The seven candidate interstellar particles are diverse in elemental composition, crystal structure, and size. The presence of crystalline grains and multiple iron-bearing phases, including sulfide, in some particles indicates that individual interstellar particles diverge from any one representative model of interstellar dust inferred from astronomical observations and theory.