Darwin R. Reyes

Microwave dielectric heating of fluids in an integrated microfluidic device

The ability to selectively and precisely control the temperature of fluid volumes ranging from a few microliters to sub-nanoliters in microfluidic networks is vital for a wide range of applications in micro total analysis systems (μTAS). In this work, we characterize and model the performance of a thin film microwave transmission line integrated with a microfluidic channel to heat fluids with relevant buffer salt concentrations over a wide range of frequencies. A microchannel fabricated in poly(dimethylsiloxane) (PDMS) is aligned with a thin film microwave transmission line in a coplanar waveguide (CPW) configuration. The electromagnetic fields localized in the gap between the signal and ground lines of the transmission line dielectrically heat the fluid in the selected region of the microchannel. Microwave S-parameter measurements and optical fluorescence-based temperature measurements are used with a theoretical model developed based on classical microwave absorption theory to fully characterize the temperature rise of the fluid. We observe a 0.95 °C mW−1 temperature rise at 15 GHz and confirm that the temperature rise of the fluid is predominantly due to microwave dielectric heating.

Quantum Dot FRET-Based Probes in Thin Films Grown in Microfluidic Channels

This paper describes the development of new fluorescence resonance energy transfer (FRET)-based quantum dot probes for proteolytic activity. The CdSe/ZnS quantum dots are incorporated into a thin polymeric film, which is prepared by layer-by-layer deposition of alternately charged polyelectrolytes. The quantum dots, which serve as fluorescent donors, are separated from rhodamine acceptor molecules, which are covalently attached to the film surface by a varying number of polyelectrolyte layers. When excited with visible light, the emission color of the polyelectrolyte multilayer film appears orange due to FRET between the quantum dots and molecular acceptors. The emission color changes to green when the rhodamine molecules are removed from the surface by enzymatic cleavage. The new probe design enables the use of quantum dots in bioassays, in this study for real-time monitoring of trypsin activity, while alleviating concerns about their potential toxicity. Application of these quantum dot FRET-based probes in microfluidic channels enables bioanalysis of volume-limited samples and single-cell studies in an in vivo-like environment.

Polyelectrolyte Multilayer-Treated Electrodes for Real-Time Electronic Sensing of Cell Proliferation.

We report on the use of polyelectrolyte multilayer (PEM) coatings as a non-biological surface preparation to facilitate uniform cell attachment and growth on patterned thin-film gold (Au) electrodes on glass for impedance-based measurements. Extracellular matrix (ECM) proteins are commonly utilized as cell adhesion promoters for electrodes; however, they exhibit degradation over time, thereby imposing limitations on the duration of conductance-based biosensor experiments. The motivation for the use of PEM coatings arises from their long-term surface stability as promoters for cell attachment, patterning, and culture. In this work, a cell proliferation monitoring device was fabricated. It consisted of thin-film Au electrodes deposited with a titanium-tungsten (TiW) adhesion layer that were patterned on a glass substrate and passivated to create active electrode areas. The electrode surfaces were then treated with a poly(ethyleneimine) (PEI) anchoring layer and subsequent bilayers of sodium poly(styrene sulfonate) (PSS) and poly(allylamine hydrochloride) (PAH). NIH-3T3 mouse embryonic fibroblast cells were cultured on the device, observed by optical microscopy, and showed uniform growth characteristics similar to those observed on a traditional polystyrene cell culture dish. The optical observations were correlated to electrical measurements on the PEM-treated electrodes, which exhibited a rise in impedance with cell proliferation and stabilized to an approximate 15 % increase as the culture approached confluency. In conclusion, cells proliferate uniformly over gold and glass PEM-treated surfaces, making them useful for continuous impedance-based, real-time monitoring of cell proliferation and for the determination of cell growth rate in cellular assays.

Hybrid cell adhesive material for instant dielectrophoretic cell trapping and long-term cell function assessment.

Dielectrophoresis (DEP) for cell manipulation has focused, for the most part, on approaches for separation/enrichment of cells of interest. Advancements in cell positioning and immobilization onto substrates for cell culture, either as single cells or as cell aggregates, has benefited from the intensified research efforts in DEP (electrokinetic) manipulation. However, there has yet to be a DEP approach that provides the conditions for cell manipulation while promoting cell function processes such as cell differentiation. Here we present the first demonstration of a system that combines DEP with a hybrid cell adhesive material (hCAM) to allow for cell entrapment and cell function, as demonstrated by cell differentiation into neuronlike cells (NLCs). The hCAM, comprised of polyelectrolytes and fibronectin, was engineered to function as an instantaneous cell adhesive surface after DEP manipulation and to support long-term cell function (cell proliferation, induction, and differentiation). Pluripotent P19 mouse embryonal carcinoma cells flowing within a microchannel were attracted to the DEP electrode surface and remained adhered onto the hCAM coating under a fluid flow field after the DEP forces were removed. Cells remained viable after DEP manipulation for up to 8 d, during which time the P19 cells were induced to differentiate into NLCs. This approach could have further applications in areas such as cell-cell communication, three-dimensional cell aggregates to create cell microenvironments, and cell cocultures.

Dielectrophoretic trapping of P19 cells on indium tin oxide based microelectrode arrays

A microfabricated device comprised of a microelectrode array (MEA) and a microfluidic channel is presented here for the purpose of trapping cells using positive dielectrophoresis (DEP). Transparent indium tin oxide (ITO) electrodes are patterned in an array of electrode pairs. A microfluidic channel made up of polydimethylsiloxane (PDMS) is then attached on top of the electrode array. DEP is used to trap P19 cells at specific positions on the ITO electrode array within the PDMS channel. Our method provides exact positioning of cells and better cell access. We show here the design and results on cell trapping with this novel microelectrode array.

Microfluidic based contactless dielectrophoretic device: Modeling and analysis

While there have been many attempts at patterning cells onto substrates, a reliable method for trapping cell clusters and forming cell arrays in a predefined geometry remains to be demonstrated. We intend to develop a multielectrode array platform to initially trap cells via dielectrophoresis (DEP) and to later measure their electrical activity. As a first step toward that objective, here we present an interdigitated microfabricated comb structure. We designed an optimal insulation layer via finite element modeling for maximum dielectrophoretic field strength in solution and minimal cell damage. The microfabricated structure was combined with a microfluidic channel to vertically constrain cell position. With the objective of capturing cells onto the substrate, we here show that there is an optimal thickness of dielectric which limits electrolysis in solution and still allows for sufficient dielectrophoretic force on the cells to pull them onto the surface.