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.

A simple and reliable microfabrication process for a programmable microvalve array.

We describe our reliable methodology for fabricating a complex programmable microvalve array (PMA) and its integration with a glass microcapillary electrophoresis chip. This methodology is applicable to any device that requires multilayered PDMS, multiple alignment processes, selective PDMS bonding, and multilayered integration with downstream sensing systems. Along with the detailed step-by-step process, we discuss essential quality assurance checks that can be performed throughout fabrication to assist in troubleshooting and maximizing chip yield.•Comprehensive instructions for designing and fabricating a programmable microvalve array.•Selective bonding of PDMS and glass by microcontact printing.•Numerous quality control procedures to boost chip yield.

Microvalve array fabrication using selective PDMS (polydimethylsiloxane) bonding through Perfluorooctyl-trichlorosilane passivation for long-term space exploration.

To improve the versatility and robustness of microfluidic analytical devices for space exploration, a programmable microfluidic array (PMA) has been implemented to support a variety of missions. When designing a PMA, normally closed valves are advantageous to avoid cross contamination and leaking. However, a stable fabrication method is required to prevent these valves from sticking and bonding over time. This work presents how polydimethylsiloxane (PDMS) can be bonded selectively using chemical passivation to overcome PDMS sticking issue during long-term space exploration. First, on a PDMS stamp, the vaporized perfluorooctyl-trichlorosilane (PFTCS) are deposited under - 80 kPa and 150 °C conditions. The PFTCS was then transferred onto PDMS or glass substrates by controlling temperature and time and 15 min at 150 °C provides the optimal PFTCS transfer for selective bonding. With these characterized parameters, we successfully demonstrated the fabrication of PMA to support long-term space missions. To estimate the stability of the stamped PFTCS, a PMA has been tested regularly for three years and no stiction or performance alteration was observed. A flight test has been done with a Cessaroni L1395 rocket for high g-force and vibration test and there is no difference on PMA performance after exposure of launch and landing conditions. This work shows promise as a simple and robust technique that will expand the stability and capability of PMA for space exploration.

A fully integrated isotachophoresis with a programmable microfluidic platform.

Conventional isotachophoresis (ITP) can be used for pre-concentration of a single analyte, but preconcentration of multiple analytes is time consuming due to handling and washing steps required for the extensive buffer optimization procedure. In this work, we present a programmable microfluidic platform (PMP) to demonstrate fully automated optimization of ITP of multiple analytes. By interfacing a PMP with ITP, buffer selection and repetitive ITP procedures were automated. Using lifting-gate microvalve technology, a PMP consisting of a two-dimensional microvalve array was designed and fabricated for seamless integration with an ITP chip. The microvalve array was used for basic liquid manipulation such as metering, mixing, selecting, delivering, and washing procedures to prime and run ITP. Initially, the performances of the PMP and ITP channel were validated individually by estimating volume per pumping cycle and preconcentrating Alexa Fluor 594 with appropriate trailing (TE) and leading (LE) buffers, respectively. After confirming basic functions, autonomous ITP was demonstrated using multiple analytes (Pacific blue, Alexa Fluor 594, and Alexa Fluor 488). The optimal buffer combination was was determined by performing multiple ITP runs with three different TEs (borate, HEPES, and phosphate buffers) and three different concentrations of Tris-HCl for the LE. We found that 40 mM borate and 100 mM Tris-HCl successfully preconcentrated all analytes during a single ITP run. The integrated PMP-ITP system can simplify overall buffer selection and validation procedures for various biological and chemical target samples. Furthermore, by incorporating analytical tools that interconnect with the PMP, it can provide high sample concentrations to aid in downstream analysis.

Characterization and Optimization of Isotachophoresis Parameters for Pacific Blue Succinimidyl Ester Dye on a PDMS Microfluidic Chip.

Isotachophoresis (ITP) for Pacific Blue (PB) dye using a polydimethylsiloxane (PDMS) microfluidic chip is developed and characterized by determining the types and concentrations of electrolytes, the ITP duration, and the electric field density. Among candidate buffers for the trailing electrolyte (TE) and leading electrolyte (LE), 40 mM borate buffer (pH 9) and 200 mM trisaminomethane hydrochloride (Tris-HCl) (pH 8) were selected to obtain the maximum preconcentration and resolution of the PB bands, respectively. With the selected TE and LE buffers, further optimization was performed to determine the electric field (EF) density and the ITP duration. These ITP parameters showed a 20-170,000 preconcentration ratio from initial PB concentrations of 10 nM-100 fM. Further demonstration was implemented to preconcentrate PB-conjugated lactate dehydrogenase (LDH) using the PDMS microfluidic chip. By utilizing the quenching nature of PB-LDH conjugation, we were able to identify concentrations of LDH as low as 10 ng/mL. This simple PDMS microfluidic chip-based ITP for PB preconcentration enables highly sensitive biological and chemical analyses by coupling with various downstream detection systems.

A microfluidic cardiac flow profile generator for studying the effect of shear stress on valvular endothelial cells.

To precisely investigate the mechanobiological responses of valvular endothelial cells, we developed a microfluidic flow profile generator using a pneumatically-actuated micropump consisting of microvalves of various sizes. By controlling the closing pressures and the actuation times of these microvalves, we modulated the magnitude and frequency of the shear stress to mimic mitral and aortic inflow profiles with frequencies in the range of 0.8-2 Hz and shear stresses up to 20 dyn cm-2. To demonstrate this flow profile generator, aortic inflow with an average of 5.9 dyn cm-2 shear stress at a frequency of 1.2 Hz with a Reynolds number of 2.75, a Womersley number of 0.27, and an oscillatory shear index (OSI) value of 0.2 was applied to porcine aortic valvular endothelial cells (PAVECs) for mechanobiological studies. The cell alignment, cell elongation, and alpha-smooth muscle actin (αSMA) expression of PAVECs under perfusion, steady flow, and aortic inflow conditions were analyzed to determine their shear-induced cell migration and trans-differentiation. In this morphological and immunocytochemical study, we found that the PAVECs elongated and aligned themselves perpendicular to the directions of the steady flow and the aortic inflow. In contrast, under perfusion with a fluidic shear stress of 0.47 dyn cm-2, the PAVECs elongated and aligned themselves parallel to the direction of flow. The PAVECs exposed to the aortic inflow upregulated their αSMA-protein expression to a greater degree than those exposed to perfusion and steady flow. By comparing these results to those of previous studies of pulsatile flow, we also found that the ratio of positive to negative shear stress plays an important role in determining PAVECs' trans-differentiation and adaptation to flow. This microfluidic cardiac flow profile generator will enable future valvular mechanobiological studies to determine the roles of magnitude and frequency of shear stresses.

A microengineered human corneal epithelium-on-a-chip for eye drops mass transport evaluation.

Animals are commonly used for pharmacokinetic studies which are the most frequent events tested during ocular drug development and preclinical evaluation. Inaccuracy, cost, and ethical criticism in these tests have created a need to construct an in vitro model for studying corneal constraints. In this work, a porous membrane embedded microfluidic platform is fabricated that separates a chip into an apical and basal side. After functionalizing the membrane surface with fibronectin, the membrane's mechanical and surface properties are measured to ensure correct modeling of in vivo characteristics. Immortalized human corneal epithelial cells are cultured on the membrane to create a microengineered corneal epithelium-on-a-chip (cornea chip) that is validated with experiments designed to test the barrier properties of the human corneal epithelium construct using model drugs. A pulsatile flow model is used that closely mimics the ocular precorneal constraints and is reasonable for permeability analysis that models in vivo conditions. This model can be used for preclinical evaluations of potential therapeutic drugs and to mimic the environment of the human cornea.

Microengineered biomimetic ocular models for ophthalmological drug development.

Current ophthalmological drug discovery and testing methods have limitations and concerns regarding reliability, ethicality, and applicability. These drawbacks can be mitigated by developing biomimetic eye models through mathematical and experimental methods which are often referred to as "eye-on-a-chip" or "eye chip". These eye chip technologies emulate ocular physiology, anatomy, and microenvironmental conditions. Such models enable understanding of the fundamental biology, pharmacology, and toxicology mechanisms by investigating the pharmacokinetics and pharmacodynamics of various candidate drugs under ocular anatomical and physiological conditions without animal models. This review provides a comprehensive overview of the latest advances in theoretical and in vitro experimental models of the anterior segment of the eye and its microenvironment, including eye motions and tear film dynamics. The current state of ocular modeling and simulation from predictive models to experimental models is discussed in detail with their advantages and limitations. The potential for future eye chip models to expedite new ophthalmic drug discoveries is also discussed.