RESUMO
Functionalized trialkoxysilanes are widely used to modify the surface properties of materials and devices. It will be shown that the photoinitiated radical-based thiol-ene "click" reaction provides a simple and efficient route to diverse trialkoxysilanes. A total of 15 trialkoxysilanes were synthesized by reacting either alkenes with 3-mercaptopropyltrialkoxysilane or thiols with allyltrialkoxysilanes in the presence of a photoinitiator. The functionalized trialkoxysilanes were obtained in quantitative to near-quantitative yields with high purity. The photochemical reactions can be run neat in standard borosilicate glassware using a low power 15-W blacklight. A wide range of functional groups is tolerated in this approach, and even complex alkenes click with the silane precursors. To demonstrate that these silanes can be used as surface coating agents, several were reacted with iron oxide superparamagnetic nanoparticles and the loadings quantified. The photoinitiated thiol-ene reaction thus offers a facile and efficient method for preparing surface-active functional trialkoxysilanes.
RESUMO
A droplet-based (digital) microfluidics platform has been developed to prepare and purify protein samples for measurement by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). Liquid droplets are moved in air by sequentially applying an electric potential to an array of electrodes patterned beneath a hydrophobic dielectric layer. We show that a complete integrated sequence of protein processing steps can be performed on this platform, including disulfide reduction, alkylation, and enzymatic digestion, followed by cocrystallization with a MALDI matrix and analysis of the sample in situ by MALDI-MS. Proteins carbonic anhydrase, cytochrome c, and ubiquitin were used to demonstrate the digestion and postdigestion steps; insulin, serum albumin, and lysozyme were used to illustrate the complete sequence of protein processing steps available with the platform. Several functional improvements in the platform are reported, notably, the incorporation of acetonitrile in the protein droplets to facilitate movement, and patterning the device surfaces to optimize sample crystallization. The method is fast, simple, repeatable, and results in lower reagent consumption and sample loss than conventional techniques for proteomics sample preparation.
Assuntos
Microfluídica , Processamento de Proteína Pós-Traducional , Proteínas/metabolismo , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , AlquilaçãoRESUMO
Localized heating of droplets on an electrowetting-on-dielectric (EWOD) chip has been implemented and shown to accelerate trypsin digestion reaction rates, sample drying, and matrix crystallization for matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS). Achieving this involved extending the functionality of previous EWOD droplet-based techniques by developing a multifunctional electrode with closed-loop temperature control, while minimizing overall system complexity and addressing challenges associated with rapid evaporation. For the EWOD chip design, we discuss the performance of multifunctional surface electrodes for actuation, localized Joule heating, and thermistic temperature sensing. Furthermore, a hydrophilic pattern is formed in the multifunctional electrode to control the location of an evaporating droplet on the electrode. To demonstrate the capabilities and limitations of this technique, we performed three experiments and measured the results using MALDI-MS: (i) insulin disulfide reductions in dithiothreitol (DTT) over a range of heater temperatures (22-70 °C) to show how reaction rates can be affected by thermal control, (ii) insulin disulfide reductions at 130 °C in dimethyl sulfoxide (DMSO) to demonstrate a reaction in a high boiling point solvent, and (iii) tryptic digestions of cytochrome c at 22 and 40 °C to show that heated droplets can yield reasonably higher peptide sequence coverage than unheated droplets. Although they do not decouple the effects of changing temperatures and concentrations, these experiments verified that thermal cycling by EWOD electrodes accelerates reaction rates in liquid droplets in air.
Assuntos
Citocromos c/metabolismo , Insulina/química , Técnicas Analíticas Microfluídicas/métodos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Dimetil Sulfóxido/química , Eletrodos , Eletroumectação/métodos , Insulina/metabolismo , Oxirredução , Temperatura , Tripsina/metabolismoRESUMO
The organic oxidant TEMPO (2,2,4,4-tetramethylpiperdine-1-oxyl) was immobilized on iron oxide (Fe(3)O(4)) superparamagnetic nanoparticles by employing strong metal-oxide chelating phosphonates and azide/alkyne "click" chemistry. This simple preparation yields recyclable TEMPO-coated nanoparticles with good TEMPO loadings. They have excellent magnetic response and efficiently catalyze the oxidation of a wide range of primary and secondary alcohols to aldehydes, ketones, and lactones under either aerobic acidic Mn(II)/Cu(II) oxidizing Minisci conditions, or basic NaOCl Anelli conditions. The nanoparticles could be recycled more than 20 times under the Minisci conditions and up to eight times under the Anelli conditions with good to excellent substrate conversions and product selectivities. Immobilization of the catalyst through a phosphonate linkage allows the particles to withstand acidic oxidizing environments with minimal catalyst leaching. Clicking TEMPO to the phosphonate prior to phosphonate immobilization, rather than after, ensures the clicked catalyst is the only species on the particle surface. This facilitates quantification of the catalyst loading. The stability of the phosphonate linker and simplicity of this catalyst immobilization method make this an attractive approach for tethering catalysts to oxide supports, creating magnetically separable catalysts that can be used under neutral or acidic conditions.
Assuntos
Álcoois/química , Óxido Ferroso-Férrico/química , Nanopartículas/química , Catálise , Química Click , Estrutura Molecular , OxirreduçãoRESUMO
Porous and hollow particles are widely used in pharmaceuticals, as solid phases for chromatography, as catalyst supports, in bioanalytical assays and medical diagnostics, and in many other applications. By controlling size, shape, and chemistry, it is possible to tune the physical and chemical properties of the particles. In some applications of millimeter-scale hollow shells, such as in high energy density physics, controlling the shell thickness uniformity (concentricity) and roundness (sphericity) becomes particularly important. In this work, we demonstrate the feasibility of using electric field-driven droplet centering to form highly spherical and concentric polymerizable double emulsion (DE) droplets that can be subsequently photopolymerized into polymer shells. Specifically, when placed under the influence of an â¼6 × 10(4) V(rms)/m field at 20 MHz, DE droplets, consisting of silicone oil as the inner droplet and tripropylene glycol diacrylate with a photoinitiator in N,N-dimethylacetamide as the outer droplet, suspended in ambient silicone oil, were found to undergo electric field-driven centering into droplets with ≥98% sphericity and â¼98% concentricity. The centered DE droplets were photopolymerized in the presence of the electric field. The high degrees of sphericity and concentricity were maintained in the polymerized particles. The poly(propylene glycol diacrylate) capsules are just within the sphericity requirements needed for inertial confinement fusion experiments. They were slightly outside the concentricity requirement. These results suggest that electric field-driven centering and polymerization of double emulsions could be very useful for synthesizing hollow polymer particles for applications in high energy density physics experiments and other applications of concentric polymer shells.
Assuntos
Resinas Acrílicas/química , Eletricidade , Polimerização , Emulsões , Estudos de Viabilidade , Polimerização/efeitos da radiação , Fatores de Tempo , Raios UltravioletaRESUMO
We demonstrate the first programmed transport of live yeast (Saccharomyces cerevisiae) and a zebrafish embryo (Danio rerio) within droplets in a two-plate digital microfluidic device. The yeast remained viable after transport, and the actuated droplets left no yeast behind. A zebrafish embryo transported 2 hours after fertilization developed normally and hatched. Dechorionation was demonstrated by mixing a droplet of digestive reagent droplet with a droplet containing the embryo. These results demonstrate the potential for using a droplet microfluidic device as an alternative to microwell plates for yeast and zebrafish assays.
Assuntos
Técnicas Analíticas Microfluídicas/métodos , Preservação Biológica/métodos , Saccharomyces cerevisiae/citologia , Peixe-Zebra/embriologia , Animais , Técnicas de Cultura Embrionária , Movimento (Física)RESUMO
Both conducting and insulating liquids can be actuated in two-plate droplet ("digital") microfluidic devices. Droplet movement is accomplished by applying a voltage across electrodes patterned beneath the dielectric-coated top and bottom plates. This report presents a general electromechanical model for calculating the forces on insulating and conducting liquids in two-plate devices. The devices are modeled as an equivalent circuit in which the dielectric layers and ambient medium (air or oil) are described as capacitors, while the liquid being actuated is described as a resistor and capacitor in parallel. The experimental variables are the thickness and dielectric constant of each layer in the device, the gap between plates, the applied voltage and frequency, and the conductivity of the liquid. The model has been used to calculate the total force acting on droplets of liquids that have been studied experimentally, and to explain the relative ease with which liquids of different conductivities can be actuated. The contributions of the electrowetting (EW) and dielectrophoretic (DEP) forces to droplet actuation have also been calculated. While for conductive liquids the EW force dominates, for dielectric liquids, both DEP and EW contribute, and the DEP force may dominate. The general utility of the model is that it can be used to predict the operating conditions needed to actuate particular liquids in devices of known geometry, and to optimize the design and operating conditions to enable movement of virtually any liquid.
Assuntos
Eletroquímica/instrumentação , Sistemas Microeletromecânicos/instrumentação , Microeletrodos , Técnicas Analíticas Microfluídicas/instrumentação , Modelos Químicos , Soluções/química , Soluções/isolamento & purificação , Simulação por Computador , Desenho Assistido por Computador , Desenho de Equipamento , Análise de Falha de Equipamento , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
The advancement of underrepresented minority and women PhD students to elite postdoctoral and faculty positions in the STEM fields continues to lag that of majority males, despite decades of efforts to mitigate bias and increase opportunities for students from diverse backgrounds. In 2015, the National Science Foundation Alliance for Graduate Education and the Professoriate (NSF AGEP) California Alliance (Berkeley, Caltech, Stanford, UCLA) conducted a wide-ranging survey of graduate students across the mathematical, physical, engineering, and computer sciences in order to identify levers to improve the success of PhD students, and, in time, improve diversity in STEM leadership positions, especially the professoriate. The survey data were interpreted via path analysis, a method that identifies significant relationships, both direct and indirect, among various factors and outcomes of interest. We investigated two important outcomes: publication rates, which largely determine a new PhD student's competitiveness in the academic marketplace, and subjective well-being. Women and minority students who perceived that they were well-prepared for their graduate courses and accepted by their colleagues (faculty and fellow students), and who experienced well-articulated and structured PhD programs, were most likely to publish at rates comparable to their male majority peers. Women PhD students experienced significantly higher levels of distress than their male peers, both majority and minority, while both women and minority student distress levels were mitigated by clearly-articulated expectations, perceiving that they were well-prepared for graduate level courses, and feeling accepted by their colleagues. It is unclear whether higher levels of distress in women students is related directly to their experiences in their STEM PhD programs. The findings suggest that mitigating factors that negatively affect diversity should not, in principle, require the investment of large resources, but rather requires attention to the local culture and structure of individual STEM PhD programs.
Assuntos
Educação de Pós-Graduação , Grupos Minoritários , Ciência/educação , Estudantes , Direitos da Mulher , California , Educação de Pós-Graduação/tendências , Engenharia/educação , Feminino , Humanos , Masculino , Matemática/educação , Grupos Minoritários/psicologia , Editoração , Estudantes/psicologia , Inquéritos e Questionários , Tecnologia/educaçãoRESUMO
To realize multiplexed sample preparation on a digital microfluidic chip for high-throughput Matrix Assisted Laser Desorption/Ionization Mass Spectrometry (MALDI-MS), several fluidic functions need to be integrated. These include the generation of multiple droplets from a reservoir and parallel in-line sample purification. In this paper, we develop two critical new functions in handling protein solutions and standard proteomic reagents with electrowetting-on-dielectric (EWOD) actuation, leading to an integrated chip for multiplexed sample preparation for MALDI-MS. The first is a voltage sequence designed to generate a series of droplets from each of the three reservoirs--proteomic sample, rinsing fluid, and MALDI reagents. It is the first time that proteomic reagents have been dispensed using EWOD in an air (as opposed to oil) environment. The second is a box-in-box electrode pattern developed to allow droplet passing over dried sample spots, making the process of in-line sample purification robust for parallel processing. As a result, parallel processing of multiple sample droplets is demonstrated on the integrated EWOD-MALDI-MS chip, an important step towards high-throughput MALDI-MS. The MS results, collected directly from the integrated devices, are of good quality, suggesting that the tedious process of sample preparation can be automated on-chip for MALDI-MS applications as well as other high-throughput proteomics applications.
Assuntos
Microfluídica/instrumentação , Microfluídica/métodos , Proteínas/análise , Proteômica/métodos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Angiotensina II/análise , Insulina/análise , Água/químicaRESUMO
In droplet-based ("digital") microfluidics, liquid droplets in contact with dielectric surfaces are created, moved, merged and mixed by applying AC or DC potentials across electrodes patterned beneath the dielectric. We show for the first time that it is possible to manipulate droplets of organic solvents, ionic liquids, and aqueous surfactant solutions in air by these mechanisms using only modest voltages (<100 V) and frequencies (<10 kHz). The feasibility of moving any liquid can be predicted empirically from its frequency-dependent complex permittivity, epsilon*. The threshold for droplet actuation in air with our two-plate device configuration is /epsilon*/>8x10(-11). The mechanistic implications of these results are discussed, along with the greatly expanded range of applications for digital microfluidics that these results suggest are now feasible.
Assuntos
Estudos de Viabilidade , Microfluídica/métodos , Soluções/química , Solventes/química , Água/química , Algoritmos , Concentração de Íons de Hidrogênio , Íons , Compostos Orgânicos/químicaRESUMO
Cell invasion is a key process in tissue growth, wound healing, and tumor progression. Most invasion assays examine cells cultured in adherent monolayers, which fail to recapitulate the three-dimensional nuances of the tissue microenvironment. Multicellular cell spheroids have a three-dimensional (3D) morphology and mimic the intercellular interactions found in tissues in vivo, thus providing a more physiologically relevant model for studying the tissue microenvironment and processes such as cell invasion. Spheroid-based invasion assays often require tedious, manually intensive handling protocols or the use of robotic liquid handling systems, which can be expensive to acquire, operate, and maintain. Here we describe a digital microfluidic (DµF) platform that enables formation of spheroids by the hanging drop method, encapsulation of the spheroids in collagen, and the exposure of spheroids to migration-modulating agents. Collagen sol-gel solutions up to 4 mg mL(-1), which form gels with elastic moduli up to â¼50 kPa, can be manipulated on the device. In situ spheroid migration assays show that cells from human fibroblast spheroids exhibit invasion into collagen gels, which can be either enhanced or inhibited by the delivery of exogenous migration modulating agents. Exposing fibroblast spheroids to spheroid secretions from colon cancer spheroids resulted in a >100% increase in fibroblast invasion into the collagen gel, consistent with the cancer-associated fibroblast phenotype. These data show that DµF can be used to automate the liquid handling protocols for spheroid-based invasion assays and create a cell invasion model that mimics the tissue microenvironment more closely than two-dimensional culturing techniques do. A DµF platform that facilitates the creation and assaying of 3D in vitro tissue models has the potential to make automated 3D cell-based assays more accessible to researchers in the life sciences.
Assuntos
Técnicas Citológicas/instrumentação , Dispositivos Lab-On-A-Chip , Esferoides Celulares/citologia , Células HT29 , Homeostase , Humanos , Esferoides Celulares/metabolismo , Esferoides Celulares/patologiaRESUMO
Cell spheroids are multicellular aggregates, grown in vitro, that mimic the three-dimensional morphology of physiological tissues. Although there are numerous benefits to using spheroids in cell-based assays, the adoption of spheroids in routine biomedical research has been limited, in part, by the tedious workflow associated with spheroid formation and analysis. Here we describe a digital microfluidic platform that has been developed to automate liquid-handling protocols for the formation, maintenance, and analysis of multicellular spheroids in hanging drop culture. We show that droplets of liquid can be added to and extracted from through-holes, or "wells," and fabricated in the bottom plate of a digital microfluidic device, enabling the formation and assaying of hanging drops. Using this digital microfluidic platform, spheroids of mouse mesenchymal stem cells were formed and maintained in situ for 72 h, exhibiting good viability (>90%) and size uniformity (% coefficient of variation <10% intraexperiment, <20% interexperiment). A proof-of-principle drug screen was performed on human colorectal adenocarcinoma spheroids to demonstrate the ability to recapitulate physiologically relevant phenomena such as insulin-induced drug resistance. With automatable and flexible liquid handling, and a wide range of in situ sample preparation and analysis capabilities, the digital microfluidic platform provides a viable tool for automating cell spheroid culture and analysis.
Assuntos
Adenocarcinoma/tratamento farmacológico , Dispositivos Lab-On-A-Chip/estatística & dados numéricos , Células-Tronco Mesenquimais/citologia , Microfluídica/métodos , Esferoides Celulares/patologia , Animais , Automação Laboratorial , Linhagem Celular Tumoral , Sobrevivência Celular , Resistência a Medicamentos , Ensaios de Seleção de Medicamentos Antitumorais/métodos , Humanos , Camundongos , Técnicas de Cultura de Órgãos , Esferoides Celulares/efeitos dos fármacos , Engenharia Tecidual/métodosRESUMO
Electrowetting-on-dielectric (EWOD) is a new method for moving liquids in biofluidic chips through electrical modification of the surface hydrophobicity. EWOD-based devices are reconfigurable, have low power requirements, and can handle neutral and charged analytes, as well as particulates. We show that biomolecular adsorption in EWOD is minimized by limiting the time during which no potential is applied and through choice of solution pH and electrode polarity. The same approach may be useful for controlling biomolecular adsorption in other applications of hydrophobic dielectric materials. These results demonstrate the feasibility of implementing EWOD for fluid actuation in biofluidic chips.
RESUMO
Natural tissues are composed of functionally diverse cell types that are organized in spatially complex arrangements. Organogenesis of complex tissues requires a coordinated sequential transformation process, with individual stages involving time-dependent expression of cell-cell, cell-matrix, and cell-signal interactions in three dimensions. The common theme of temporal-spatial patterning of these cellular interactions is also observed in other physiological processes, such as growth and development, wound healing, and tumor migration. The "precursor tissue analog" (PTA) applies the temporal-spatial patterning theme to tissue engineering. The goal of PTA in tissue engineering is not to fabricate the final transplantable tissue but rather to guide the dynamic organization, maturation, and remodeling leading to the formation of normal and functional tissues. We describe the critical design principles of PTA. First, structural, mechanical, and physiological requirements of the PTA as a temporary scaffold must be met by a fabrication method with flexibility. The fabrication potential incorporating biological materials such as living cells and plasmid DNA has been addressed. Second, the PTA concept is considered suitable for future tissue engineering in light of the use of undifferentiated stem cells, and may possess a capability to guide stem cells toward diverse differentiation characteristics in situ. To this end, the behavior of the engineered cell and tissue must be monitored in detail. The development of a practical phenotype monitoring system such as a DNA microarray may be integral to the fabrication strategies of PTA. Third, the microtopographical and microenvironmental control on the liquid-solid interaction may lead to a critical design for PTA to provide soluble factors, nutrients, and gases to the cells embedded within the scaffold. We suggest that the level set numerical simulation method may be utilized to engineer the consistent circulation of bioactive liquid throughout the PTA microenvironment.
Assuntos
Diferenciação Celular/fisiologia , Células-Tronco/fisiologia , Engenharia Tecidual , Animais , Materiais Biocompatíveis , Humanos , Análise de Sequência com Séries de Oligonucleotídeos , ImpressãoRESUMO
A droplet (digital) microfluidic device has been developed that enables complete protein sample preparation for MALDI-MS analysis. Protein solution dispensing, disulfide bond reduction and alkylation, tryptic digestion, sample crystallization, and mass spectrometric analysis are all performed on a single device without the need for any ex situ sample purification. Fluorinated solvents are used as an alternative to surfactants to facilitate droplet movement and limit protein adsorption onto the device surface. The fluorinated solvent is removed by evaporation and so does not interfere with the MALDI-MS analysis. Adding a small amount of perfluorooctanoic acid to the MALDI matrix solution improves the yield, quality and consistency of the protein-matrix co-crystals, reducing the need for extensive 'sweet spot' searching and improving the spectral signal-to-noise ratio. These innovations are demonstrated in the complete processing and MALDI-MS analysis of lysozyme and cytochrome c. Because all of the sample processing steps and analysis can be performed on a single digital microfluidic device without the need for ex situ sample handling, higher throughput can be obtained in proteomics applications. More generally, the results presented here suggest that fluorinated liquids could also be used to minimize protein adsorption and improve crystallization in other types of lab-on-a-chip devices and applications.
Assuntos
Citocromos c/química , Técnicas Analíticas Microfluídicas , Muramidase/química , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Tensoativos/química , Caprilatos/química , Cristalização , Citocromos c/metabolismo , Fluorocarbonos/química , Halogenação , Muramidase/metabolismo , Peptídeos/análise , Razão Sinal-Ruído , Tripsina/metabolismoRESUMO
An in-line sample purification method for MALDI-MS, which relies on the electrowetting-on-dielectric (EWOD)-based technique for digital microfluidics, is reported. In this method, a droplet containing peptides and impurities is moved by EWOD and deposited onto a Teflon-AF surface. A droplet of water is subsequently moved over the spot, where it dissolves and removes the impurities. A droplet containing MALDI matrix is then moved to the spot, which is analyzed by MALDI-MS. This purification method reduces the number of salt adduct peaks caused by low concentrations of impurities (e.g., 20 mM sodium phosphate), and reduces or eliminates the catastrophic effects of high concentrations of impurities (e.g., 8 M urea). The method was used to purify spots made by depositing multiple droplets of contaminated peptides. Spectra from the purified spots showed an increase in the S/N ratio as a function of the number of droplets deposited; when not purified, the S/N ratio remained constant regardless of the number of droplets. Finally, the method was used to purify protein digests for peptide mass fragment (PMF) searches, and was shown to be more efficient than the conventional method of purification with reversed-phase-packed pipet tips. We anticipate this new, in-line sample purification technique for EWOD-MALDI-MS will enable development of integrated high-throughput proteomics analysis methodologies.
Assuntos
Técnicas Analíticas Microfluídicas/métodos , Proteômica/métodos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Métodos Analíticos de Preparação de Amostras , Angiotensina II/análise , Metaloendopeptidases/metabolismo , Mioglobina/metabolismo , Ubiquitina/metabolismoRESUMO
The biological activity of proteins is structure dependent. In this discussion, we describe development of g-factor analysis for characterizing the secondary structure of proteins in solutions and films. In g-factor analysis, experimental circular dichroism (CD) and UV absorbance spectra are converted to dimensionless g-factor spectra by dividing the differential absorbance of circularly polarized light (AL - AR) by the UV absorbance (A) at each wavelength. The spectra can be deconvolved and the secondary structure estimated without information on the protein molecular weight, sample concentration, or sample path length. We have refined g-factor spectral acquisition and deconvolution parameters to improve the precision and accuracy of experimental g-factor spectra and the subsequent secondary structure estimations. In general, slower scan rates and longer response times are required to obtain high quality g-factor spectra than to obtain ordinary CD spectra, particularly for data at wavelengths longer than 230 nm. The spectral acquisition and deconvolution procedures have been validated for aqueous bovine serum albumin (BSA) and poly(L-proline). The secondary structures of fibronectin and laminin in buffer solutions are predicted. When BSA and poly(L-proline) form films on quartz, their secondary structures change significantly: 13% for BSA and 32% for poly(L-proline). By contrast, the secondary structure of fibronectin is the same in solution and films. The g-factor method is an easy, rapid, accurate and precise method for determining secondary structure and structural changes in protein solutions and films. Potential applications range from proteomics and structure-based drug discovery, to the design and fabrication of biosensors, biomaterials and biofluidic devices.
Assuntos
Análise Fatorial , Estrutura Secundária de Proteína , Algoritmos , Dicroísmo Circular , Cristalografia por Raios X , Fibronectinas/química , Laminina/química , Espectroscopia de Ressonância Magnética , Peptídeos/química , Proteínas/química , Soroalbumina Bovina/química , Soluções , Espectrofotometria UltravioletaRESUMO
A new technique for preparing samples for matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) is reported. The technique relies on electrowetting-on-dielectric (EWOD) to move droplets containing proteins or peptides and matrix to specific locations on an array of electrodes for analysis. Standard MALDI-MS reagents, analytes, concentrations, and recipes are demonstrated to be compatible with the technique. Mass spectra are comparable to those collected by conventional methods. Nonspecific adsorption of analytes to device surfaces is demonstrated to be negligible. The results suggest that EWOD may be a useful tool for automating sample preparation for high-throughput proteomics and other applications of MALDI-MS.