polymeric-microneedle-array-fabrication-by-photolithography

Polymeric Microneedle Array Fabrication by Photolithography

This manuscript describes the fabrication of polymeric microneedle (MN) arrays by photolithography. It involves a simple mold-free process by using a ...

Agency for Science, Technology and Research (A*STAR)

The paper presents a 3-D filter chip employing both mechanical and dielectrophoretic (DEP) filtration, and its corresponding microfabrication techniques. The device structure is similar to a classical capacitor: two planar electrodes, made from a stainless steel mesh, and bonded on both sides of a glass frame filled with round silica beads. The solution with the suspension of particles flows through both the mesh-electrodes and silica beads filter. The top stainless steel mesh (with openings of 60 mum and wires of 30 mum-thickness) provides the first stage of filtration based on mechanical trapping. A second level of filtration is based on DEP by using the nonuniformities of the electric field generated in the capacitor due to the nonuniformities of the dielectric medium. The filter can work also with DC and AC electric fields. The device was tested with yeast cells (Saccharomyces cerevisae) and achieved a maximal trapping efficiency of 75% at an applied AC voltage of 200 V and a flow rate of 0.1 mL/min, from an initial concentration of cells of 5 x 10(5) cells/mL. When the applied frequency was varieted in the range between 20 and 200 kHz, a minimal value of capture efficiency (3%) was notticed at 50 kHz, when yeast cells exhibit negative DEP and the cells are repelled in the space between the beads.

A 3-D dielectrophoretic filter chip.

Microfabricated systems provide an excellent platform for the culture of cells, and are an extremely useful tool for the investigation of cellular responses to various stimuli. Advantages offered over traditional methods include cost-effectiveness, controllability, low volume, high resolution, and sensitivity. Both biocompatible and bio-incompatible materials have been developed for use in these applications. Biocompatible materials such as PMMA or PLGA can be used directly for cell culture. However, for bio-incompatible materials such as silicon or PDMS, additional steps need to be taken to render these materials more suitable for cell adhesion and maintenance. This review describes multiple surface modification strategies to improve the biocompatibility of MEMS materials. Basic concepts of cell-biomaterial interactions, such as protein adsorption and cell adhesion are covered. Finally, the applications of these MEMS materials in Tissue Engineering are presented.

Cell culture on MEMS platforms: a review.

This paper presents a field-flow method for separating particle populations in a dielectrophoretic (DEP) chip with asymmetric electrodes under continuous flow. The structure of the DEP device (with one thick electrode that defines the walls of the microfluidic channel and one thin electrode), as well as the fabrication and characterization of the device, was previously described. A characteristic of this structure is that it generates an increased gradient of electric field in the vertical plane that can levitate the particles experiencing negative DEP. The separation method consists of trapping one population to the bottom of the microfluidic channel using positive DEP, while the other population that exhibits negative DEP is levitated and flowed out. Viable and nonviable yeast cells were used for testing of the separation method.

Dielectrophoretic field-flow method for separating particle populations in a chip with asymmetric electrodes.

Hepatotoxicity evaluation of pharmaceutical lead compounds in early stages of drug development has drawn increasing attention. Sandwiched hepatocytes exhibiting stable functions in culture represent a standard model for hepatotoxicity testing of drugs. We have developed a robust and high-throughput hepatotoxicity testing platform based on the sandwiched hepatocytes for drug screening. The platform involves a galactosylated microfabricated membrane sandwich to support cellular function through uniform and efficient mass transfer while protecting cells from excessive shear. Perfusion bioreactor further enhances mass transfer and cellular functions over long period; and hepatocytes are readily transferred to 96-well plate for high-throughput robotic liquid handling. The bioreactor design and perfusion flow rate are optimized by computational fluid dynamics simulation and experimentally. The cultured hepatocytes preserved 3D cell morphology, urea production and cytochrome p450 activity of the mature hepatocytes for 14 days. When the perfusion-cultured sandwich is transferred to 96-well plate for drug testing, the hepatocytes exhibited improved drug sensitivity and low variability in hepatotoxicity responses amongst cells transferred from different dates of perfusion culture. The platform enables robust high-throughput screening of drug candidates.

A robust high-throughput sandwich cell-based drug screening platform.

There are a plethora of approaches to construct microtissues as building blocks for the repair and regeneration of larger and complex tissues. Here we focus on various physical and chemical trapping methods for engineering three-dimensional microtissue constructs in microfluidic systems that recapitulate the in vivo tissue microstructures and functions. Advances in these in vitro tissue models have enabled various applications, including drug screening, disease or injury models, and cell-based biosensors. The future would see strides toward the mesoscale control of even finer tissue microstructures and the scaling of various designs for high throughput applications. These tools and knowledge will establish the foundation for precision engineering of complex tissues of the internal organs for biomedical applications.

Exploitation of physical and chemical constraints for three-dimensional microtissue construction in microfluidics.

We have developed a multi-channel microfluidic perfusion platform for culturing zebrafish embryos and capturing live images of various tissues and organs inside the embryo. The Fish and Chips was micro-fabricated in silicon and glass for reproducibility and accuracy of the microfluidic structure. The microfluidic platform consists of three parts: a microfluidic gradient generator, a row of eight fish tanks, in which the fish embryos are individually placed, and eight output channels. The fluidic gradient generator supports dose-dependent drug and chemical studies. A unique perfusion system ensures a uniform and constant flow of media to the fish tank while the wastes are efficiently removed. The fish tanks restrict the embryo movements, except rotationally, for live imaging of internal tissues and organs. The embryos showed developmental abnormalities under the influence of the drug valproic acid (VPA).

Fish and Chips: a microfluidic perfusion platform for monitoring zebrafish development.

This paper describes the main protocols that are used for fabricating microfluidic devices from glass and silicon. Methods for micropatterning glass and silicon are surveyed, and their limitations are discussed. Bonding methods that can be used for joining these materials are summarized and key process parameters are indicated. The paper also outlines techniques for forming electrical connections between microfluidic devices and external circuits. A framework is proposed for the synthesis of a complete glass/silicon device fabrication flow.

A practical guide for the fabrication of microfluidic devices using glass and silicon.

Synthesis of surfactant-polyelectrolyte nanoparticles was carried out in a microfluidic device with a fine control over the size and the polydispersity. An anionic polysaccharide (sodium carboxymethylcellulose, CMC) solution was focused using a cationic surfactant (dodecyl trimethylammonium bromide, DTAB) solution in a microfluidic channel at selected ratios of flow rates and reagent concentrations. The methodology ensured a controlled mixing kinetics and a uniform distribution of charges at the mixing interface. The resulting nanoparticles exhibited remarkably well-defined and repeatable size distributions, with hydrodynamic diameters tunable from 50 up to 300 nm and polydispersity index around 0.1 in most cases. Microfluidic-assisted self-assembly may be an efficient way to produce well-controlled polyelectrolyte-based nanoparticles suitable for colloidal science as well as for gene delivery applications.

Fine control over the size of surfactant-polyelectrolyte nanoparticles by hydrodynamic flow focusing.

We demonstrate here the application of electrochemical impedance spectroscopy (EIS) in microfluidic devices for label-free virus identification by means of their specific "signature" and also investigate its feasibility for titer quantitation using two basic approaches. The first one is a method based on identifying so-called "resonance" frequencies manifesting in our microdevices and monitoring their variation as a function of the virus concentration, whereas the second one relies on measuring the relative impedance variation at these "resonance" frequencies. Best results have been obtained for the highest "resonance" frequency (∼80 MHz), which we attribute to be due to both the structure of the microdevice and the extremely small size of the viruses that make their effect significant only at such frequencies. This is a simpler method of determining virus concentration in diluted solutions of purified viruses than the well-established traditional plaque assay titer estimation method, and-since it is based on frequency measurement-could potentially be more accurate.

Label-free virus identification and characterization using electrochemical impedance spectroscopy.

This review will cover the recent advances in label-free approaches to isolate and manipulate circulating tumor cells (CTCs). In essence, label-free approaches do not rely on antibodies or biological markers for labeling the cells of interest, but enrich them using the differential physical properties intrinsic to cancer and blood cells. We will discuss technologies that isolate cells based on their biomechanical and electrical properties. Label-free approaches to analyze CTCs have been recently invoked as a valid alternative to "marker-based" techniques, because classical epithelial and tumor markers are lost on some CTC populations and there is no comprehensive phenotypic definition for CTCs. We will highlight the advantages and drawbacks of these technologies and the status on their implementation in the clinics.

Label-free isolation of circulating tumor cells in microfluidic devices: Current research and perspectives.

To fabricate microneedle arrays directly off a photomask using a simple photolithographical approach and evaluate their potential for delivering collagen.

Direct microneedle array fabrication off a photomask to deliver collagen through skin.

This paper presents a microfluidic method for precise control of the size and polydispersity of surfactant-DNA nanoparticles. A mixture of surfactant and DNA dispersed in 35% ethanol is focused between two streams of pure water in a microfluidic channel. As a result, a rapid change of solvent quality takes place in the central stream, and the surfactant-bound DNA molecules undergo a fast coil-globule transition. By adjusting the concentrations of DNA and surfactant, fine-tuning of the nanoparticle size, down to a hydrodynamic diameter of 70 nm with a polydispersity index below 0.2, can be achieved with a good reproducibility.

Article	Year
A 3-D dielectrophoretic filter chip. Electrophoresis\| PubMed ID: 17330223	2007
Cell culture on MEMS platforms: a review. International journal of molecular sciences\| PubMed ID: 20054478	2009
Dielectrophoretic field-flow method for separating particle populations in a chip with asymmetric electrodes. Biomicrofluidics\| PubMed ID: 20216966	2009
A robust high-throughput sandwich cell-based drug screening platform. Biomaterials\| PubMed ID: 20971505	2011
Exploitation of physical and chemical constraints for three-dimensional microtissue construction in microfluidics. Biomicrofluidics\| PubMed ID: 21799710	2011
Fish and Chips: a microfluidic perfusion platform for monitoring zebrafish development. Lab on a chip\| PubMed ID: 22146879	2012
A practical guide for the fabrication of microfluidic devices using glass and silicon. Biomicrofluidics\| PubMed ID: 22662101	2012
Fine control over the size of surfactant-polyelectrolyte nanoparticles by hydrodynamic flow focusing. Analytical chemistry\| PubMed ID: 23713852	2013
Label-free virus identification and characterization using electrochemical impedance spectroscopy. Electrophoresis\| PubMed ID: 24285469	2014
Label-free isolation of circulating tumor cells in microfluidic devices: Current research and perspectives. Biomicrofluidics\| PubMed ID: 24403992	2013
Direct microneedle array fabrication off a photomask to deliver collagen through skin. Pharmaceutical research\| PubMed ID: 24449441	2014
On-chip controlled surfactant-DNA coil-globule transition by rapid solvent exchange using hydrodynamic flow focusing. Langmuir : the ACS journal of surfaces and colloids\| PubMed ID: 25351469	2014

Ciprian Iliescu

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