Future Missions to Titan: Scientific and Engineering Challenges

Saturn’s largest moon, Titan, has been an enigma at every stage of its exploration. For three decades after the hazy atmosphere was discovered from the ground in the 1940s, debate ensued over whether it was a thin layer of methane or a dense shield of methane and nitrogen. Voyager 1 settled the matter in favor of the latter in 1980, but the details of the thick atmosphere discovered raised even more intriguing questions about the nature of the hidden surface, and the sources of resupply of methane to the atmosphere. The simplest possibility, that an ocean of methane and its major photochemical product ethane might cover the globe, was cast in doubt by Earth-based radar studies and then eliminated by Hubble Space Telescope and adaptive optics imaging in the near-infrared from large ground-based telescopes in the 1990s. These data, however, did not reveal the complexity of the surface that Cassini-Huygens would uncover beginning in 2004. A hydrological cycle appears to exist in which methane (in concert with ethane in some processes) plays the role on Titan that water plays on Earth. Channels likely carved by liquid methane and/or ethane, lakes and seas of these materials—some rivaling or exceeding North America’s Great Lakes in size—vast equatorial dune fields of complex organics made high in the atmosphere and shaped by wind, and intriguing hints of geologic activity suggest a world with a balance of geologic and atmospheric processes that is the solar system’s best analogue to Earth. Deep underneath Titan’s dense atmosphere and active, diverse surface is an interior ocean discovered by Cassini and thought to be largely composed of liquid water. Cassini-Huygens has provided spectacular data and has enabled us to glimpse the mysterious surface of Titan. However the mission will leave us with many questions that require future missions to answer. These include determining the composition of the surface and the geographic distribution of various organic constituents. Key questions remain about the ages of surface features, specifically whether cryovolcanism and tectonism are actively ongoing or are relics of a more active past. Ammonia, circumstantially suggested to be present by a variety of different kinds of Cassini-Huygens data, has yet to be seen. Is methane out-gassing from the interior or ice crust today? Are the lakes fed primarily by rain or underground methane-ethane aquifers (more properly, “alkanofers”) and how often have heavy methane rains come to the equatorial region? We should investigate whether Titan’s surface supported vaster seas of methane in the past, and whether complex self-organizing chemical systems have come and gone in the water volcanism, or even exist in exotic form today in the high latitude lakes. The presence of a magnetic field has yet to be established. A large altitude range in the atmosphere, from 400–900 km in altitude, will remain poorly explored after Cassini. Much remains to be understood about seasonal changes of the atmosphere at all levels, and the long-term escape of constituents to space. Other than Earth, Titan is the only world in our solar system known to have standing liquids and an active “hydrologic cycle” with clouds, rains, lakes and streams. The dense atmosphere and liquid lakes on Titan’s surface can be explored with airborne platforms and landed probes, but the key aspect ensuring the success of future investigations is the conceptualization and design of instruments that are small enough to fit on the landed probes and airborne platforms, yet sophisticated enough to conduct the kinds of detailed chemical (including isotopic), physical, and structural analyses needed to investigate the history and cycling of the organic materials. In addition, they must be capable of operating at cryogenic temperatures while maintaining the integrity of the sample throughout the analytic process. Illuminating accurate chemistries also requires that the instruments and tools are not simultaneously biasing the measurements due to localized temperature increases. While the requirements for these techniques are well understood, their implementation in an extremely low temperature environment with limited mass, power and volume is acutely challenging. No such instrument systems exist today. Missions to Titan are severely limited in both mass and power because spacecraft have to travel over a billion miles to get there and require a large amount of fuel, not only to reach Titan, but to maintain the ability to maneuver when they arrive. Landed missions have additional limitations, in that they must be packaged in a sealed aeroshell for entry into Titan’s atmosphere. Increases in landed mass and volume translate to increased aeroshell mass and size, requiring even more fuel for delivery to Titan. Nevertheless, missions during which such systems and instruments could be employed range from Discovery and New Frontiers class in situ probes that might be launched in the next decade, to a full-up Flagship class mission anticipated to follow the Europa Jupiter System Mission. Capitalizing on recent breakthroughs in cryo-technologies and smart materials fabrication, we developed conceptual designs of sample acquisition systems and instruments capable of in situ operation under low temperature environments. The study included two workshops aimed at brainstorming and actively discussing a broad range of ideas and associated challenges with landing instruments on Titan, as well as more focused discussions during the intervening part of the study period. The workshops each lasted ~4 days (Monday-Thursday/Friday), included postdoctoral fellows and students in addition to the core team members, and generated active engagement from the Caltech and JPL team participants, as well as from the outside institutions. During the workshops, new instruments and sampling methodologies were identified to handle the challenges of characterizing everything from small molecules in Titan’s upper atmosphere to gross mixtures of high molecular weight complex organics in condensed phases, including atmospheric aerosols and “organic sand” in dunes, to highly dilute components in ices and lakes. To enable these advances in cryogenic instrumentation breakthroughs in a wide range of disciplines, including electronics, chemical and mechanical engineering, and materials science were identified

Fabrication and function of microfluidic devices for monitoring of in-vitro fertilization processes

Thesis (S.B.)--Massachusetts Institute of Technology, Dept. of Mechanical Engineering, 2007.Includes bibliographical references (leaf 36).The process of assistive reproduction is often a headache and heartache for those who choose to go through it. The field currently relies heavily on morphological characteristics to determine embryo health and development success, a highly unreliable method. While they appear healthy at implantation, many embryos, in reality, have poor development potential and fail to survive within the womb. Therefore, to offset the high chances of miscarriage, multiple eggs are implanted in the uterus. This has occasionally lead to multi-fetal pregnancies, which have a higher maternal mortality risk, and, in general, is more physically demanding. This thesis researches a microfluidic device that aids in the crucial stages of in vitro- fertilization. The device allows for a fertilized egg to be cultured within, and provides the ability to carefully monitor its health through a series of metabolic assays, a better indication of embryo health. This microfluidic embryo health monitoring device is comprised of two layers of channel networks. It works through passing fluids along flow channels that are driven by control channels. The control layer, when pressurized with gas, operates as valves and peristaltic pumps along the flow layer to pump and transport fluids through the flow channels. As embryonic fluids are passed through the channels, the status of the fertilized egg can be monitored with metabolic assays taken of the embryo at various detection sites.by Jin Xu.S.B

Real time optical immunosensing with flow-through porous alumina membranes

Through the presentation of analytical data from bioassay experiments, measured by polarimetry, we demonstrate for the first time a real time immunoassay within a free standing macroporous alumina membrane. The 200 nm nominal pore diameter of the membrane enables flow-through, thereby providing an ideal fluidic platform for the targeted delivery of analytes to bioreceptors immobilized on the pore walls, enabling fast sensing response times and the use of small sample volumes (<100 μL). For the immunoassay, the pore walls were first coated with the functional copolymer, copoly(DMA-NAS) using a novel coupling process, before immobilization of the allergen protein, β-lactoglobulin, by spotting. The immuno-assay then proceeded with the binding of the primary and secondary antibody cognates, rabbit anti-β-lactoglobulin and anti-rabbit IgG respectively. Through the use of streptavidin coated quantum dots as refractive index signal enhancers, a noise floor for individual measurements of 3.7 ng/mL (25 pM) was obtained, with an overall statistical, or formal assay LOD of 33.7 ng/mL (225 pM), for total assay time below 1 h

MilkGuard: Low-Cost, Polymer-based Sensor for the Detection of Escherichia coli in Donated Human Breast Milk

Breast milk, the gold standard for infant nutrition, could prevent up to 13% of child deaths worldwide. However, many mothers are unable to breastfeed due to health conditions and other factors. Because of this, a network of more than 500+ human milk banks, which collect and distribute donated breast milk to infants, have emerged worldwide. However, operational costs to ensure the safety of this milk remain time-intensive and costly. There are no existing diagnostics for rapid and on-site detection of bacterial contaminants in donated milk. Currently, many milk banks send samples to outside laboratories for bacterial culturing tests, which take 24-48 hours to receive results. In contrast, MilkGuard is an on-site detection method which ensures results in hours rather than days. To determine whether or not E.coli is present in donated milk, a drop of milk is deposited onto the sensor. If the milk is contaminated, the sensor will turn a blue color due to an enzyme-substrate reaction of the bacteria. The goal of the project is to create a cost and rapid alternative to traditional bacterial culturing testing to screen for E. coli bacteria in donated human breast milk. This will allow users to ensure that milk samples are sterile enough to provide to young infants, while also providing breast milk banks an alternative that will allow them to screen more samples in a shorter amount of time

Next generation transduction pathways for nano-bio-chip array platforms

textIn the following work, nanoparticle quantum dot (QD) fluorophores have been exploited to measure biologically relevant analytes via a miniaturized sensor ensemble to provide key diagnostic and prognostic information in a rapid, yet sensitive manner—data essential for effective treatment of many diseases including HIV/AIDS and cancer. At the heart of this “nano-bio-chip” (NBC) sensor is a modular chemical/cellular processing unit consisting of either a polycarbonate membrane filter for cell-based assays, or an agarose bead array for detection of biomarkers in serum or saliva. Two applications of the NBC sensor system are described herein, both exhibiting excellent correlation to reference methods ((R² above 0.94), with analysis times under 30 minutes and sample volumes below 50 [mu]L. First, the NBC sensor was employed for the sequestration and enumeration of T lymphocytes, cells specifically targeted by HIV, from whole blood samples. Several different conjugation methods linking QDs to recognition biomolecules were extensively characterized by biological and optical methods, with a thiol-linked secondary antibody labeling scheme yielding intense, specific signal. Using this technique, the photostability of QDs was exploited, as was the ability to simultaneously visualize different color QDs via a single light pathway, effectively reducing optical requirements by half. Further, T-cell counts were observed well below the 200/[mu]L discriminator between HIV and AIDS and across the common testing region, demonstrating the first reported example of cell counting via QDs in an enclosed, disposable device. Next, multiplexed bead-based detection of cancer protein biomarkers CEA, Her-2/Neu, and CA125 in serum and saliva was examined using a sandwich immunoassay with detecting antibodies covalently bound to QDs. This nano-based signal was amplified 30 times versus molecular fluorophores and cross talk in multiplexed experiments was less than 5%. In addition, molecular-level tuning of recognition elements (size, concentration) and agarose porosity resulted in NBC limits of detection two orders of magnitude lower than ELISA, values competitive with the most sensitive methods yet reported (0.021 ng/mL CEA). Taken together, these efforts serve to establish the valuable role of QDs in miniaturized diagnostic devices with potential for delivering biomedical information rapidly, reliably, and robustly.Chemistry and Biochemistr

Portable Diagnostics Technology Assessment for Space Missions

The changes in the scope of NASA s mission in the coming decade are profound and demand nimble, yet insightful, responses. On-board clinical and environmental diagnostics must be available for both mid-term lunar and long-term Mars exploration missions in an environment marked by scarce resources. Miniaturization has become an obvious focus. Despite solid achievements in lab-based devices, broad-based, robust tools for application in the field are not yet on the market. The confluence of rapid, wide-ranging technology evolution and internal planning needs are the impetus behind this work. This report presents an analytical tool for the ongoing evaluation of promising technology platforms based on mission- and application-specific attributes. It is not meant to assess specific devices, but rather to provide objective guidelines for a rational down-select of general categories of technology platforms. In this study, we have employed our expertise in the microgravity operation of fluidic devices, laboratory diagnostics for space applications, and terrestrial research in biochip development. A rating of the current state of technology development is presented using the present tool. Two mission scenarios are also investigated: a 30-day lunar mission using proven, tested technology in 5 years; and a 2- to 3-year mission to Mars in 10 to 15 years

Microfluidic devices interfaced to matrix-assisted laser desorption/ionization mass spectrometry for proteomics

Microfluidic interfaces were developed for off-line matrix-assisted laser desorption/ionization mass spectrometry (MALDI). Microfluidic interfaces allow samples to be manipulated on-chip and deposited onto a MALDI target plate for analysis. For this research, microfluidic culturing devices and automated digestion and deposition microfluidic chip platforms were developed for the identification of proteins. The microfluidic chip components were fabricated on a poly(methyl methacrylate), PMMA, wafer using the hot embossing method and a molding tool with structures prepared via micromilling. One of the most important components of the chip system was a trypsin microreactor. An open channel microreactor was constructed in a 100 µm wide and 100 µm deep channel with a 4 cm effective channel length. This device integrated frequently repeated steps for MALDI-based proteomics such as digestion, mixing with a matrix solution, and depositing onto a MALDI target. The microreactor provided efficient digestion of proteins at a flow rate of 1 µL/min with a residence time of approximately 24 s in the reaction channel. An electrokinetically driven microreactor was also developed using a micropost structured chip for digestion. The micropost chip had a higher digestion efficiency due to the higher surface area-to-volume ratio in the channel. Also, the electrokinetic flow eliminated the need for an external pumping system and gave a flat flow profile in the microchannel. The post microreactor consisted of a 4 cm × 200 µm × 50 µm microfluidic channel with trypsin immobilized on an array of 50 µm in diameter micropost support structures with a 50 µm edge-to-edge inter-post spacing. This micropost reactor was also used for fingerprint analysis of whole bacterial cells. The entire tryptic digestion and deposition procedure for intact bacteria took about 1 min. A contact deposition solid-phase bioreactor coupled with MALDI-TOF MS allowed for low-volume fraction deposition with a smaller spot size and a higher local concentration of the analyte. A bacterial cell-culturing chip was constructed for growing cells on-chip followed by off-line MALDI analysis. Coupling MALDI-TOF MS whole cell analysis with microfluidic culturing resulted in more consistent spectra as well as reduction of the total processing time. The microfluidic cell culturing was performed in a PMMA chip with a polydimethylsiloxane (PDMS) cover to allow gas permeation into the culture channel, which contained a 2.1 μL volume active culture chamber. After incubation of E. coli in a microfluidic culture device at 37 ℃ for 24 h, the cultured cells were analyzed with MALDI MS. Also, a microfluidic cell culture device containing continuous perfusion of culture medium was developed using a polycarbonate membrane. This microfluidic culturing format was improved with a fluidic manifold and thermostatted microheaters. Fingerprint mass spectra distinguishing E. coli strains tested were obtained after a 6 h incubation time, which was shorter compared to the 24 h incubation time using conventional culturing techniques. In addition, an enhanced identification procedure for bacteria was achieved by integrating on-chip digestion of cultured bacteria

3D Printed Microfluidic Devices

3D printing has revolutionized the microfabrication prototyping workflow over the past few years. With the recent improvements in 3D printing technologies, highly complex microfluidic devices can be fabricated via single-step, rapid, and cost-effective protocols as a promising alternative to the time consuming, costly and sophisticated traditional cleanroom fabrication. Microfluidic devices have enabled a wide range of biochemical and clinical applications, such as cancer screening, micro-physiological system engineering, high-throughput drug testing, and point-of-care diagnostics. Using 3D printing fabrication technologies, alteration of the design features is significantly easier than traditional fabrication, enabling agile iterative design and facilitating rapid prototyping. This can make microfluidic technology more accessible to researchers in various fields and accelerates innovation in the field of microfluidics. Accordingly, this Special Issue seeks to showcase research papers, short communications, and review articles that focus on novel methodological developments in 3D printing and its use for various biochemical and biomedical applications

Poly Drop

Poly Drop is a software interface to control an Open Drop digital micro-fluidics system. We obtained a hardware system from Gaudi labs. Our task was to create a Graphical User Interface that made the control of the device easier and more automated for better testing. We created software that had 3 parts: a control GUI, arduino code to control the hardware, and Image Analysis that gives the user information such as location and color of liquid drops as they move across the electrode grid of the Open Drop system. The GUI was developed using Java Swing. The communication between the GUI and the arduino was accomplished using the open source RXTX library. The image analysis portion was created using the open source OpenCV software