Rodrigo Martinez-Duarte

Microfabrication technologies in dielectrophoresis applications-A review

DEP is an established technique for particle manipulation. Although first demonstrated in the 1950s, it was not until the development of miniaturization techniques in the 1990s that DEP became a popular research field. The 1990s saw an explosion of DEP publications using microfabricated metal electrode arrays to sort a wide variety of cells. The concurrent development of microfluidics enabled devices for flow management and better understanding of the interaction between hydrodynamic and electrokinetic forces. Starting in the 2000s, alternative techniques have arisen to overcome common problems in metal‐electrode DEP, such as electrode fouling, and to increase the throughput of the system. Insulator‐based DEP and light‐induced DEP are the most significant examples. Most recently, new 3D techniques such as carbon‐electrode DEP, contactless DEP, and the use of doped PDMS have further simplified the fabrication process. The constant desire of the community to develop practical solutions has led to devices which are more user friendly, less expensive, and are capable of higher throughput. The state‐of‐the‐art of fabricating DEP devices is critically reviewed in this work. The focus is on how different fabrication techniques can boost the development of practical DEP devices to be used in different settings such as clinical cell sorting and infection diagnosis, industrial food safety, and enrichment of particle populations for drug development.

A novel approach to dielectrophoresis using carbon electrodes

Carbon‐electrode dielectrophoresis (carbon‐DEP) is demonstrated here as an alternative to more traditional DEP techniques. Carbon‐DEP combines advantages of metal‐electrode and insulator‐based DEP by using low‐cost fabrication techniques and low voltages for particle manipulation. The use of 3‐D electrodes is proved to yield significant advantages over the use of traditional planar electrodes. This paper details the fabrication of dense arrays of tall high aspect ratio carbon electrodes on a transparent fused‐silica substrate. The shrinkage of the SU‐8 structures during carbonization is characterized and a design tool for future devices is provided. Applications of carbon electrodes in DEP are then detailed and include particle positioning, high‐throughput filtering and cell focusing using positive‐DEP. Manipulated cells include Saccharomyces cerevisiae and Drosophila melanogaster. The advantages and disadvantages of carbon‐DEP are discussed at the end of this work.

On-line separation of bacterial cells by carbon-electrode dielectrophoresis.

Dielectrophoresis (DEP) represents a powerful approach to manipulate and study living cells. Hitherto, several approaches have used 2‐D DEP chips. With the aim to increase sample volume, in this study we used a 3‐D carbon‐electrode DEP chip to trap and release bacterial cells. A continuous flow was used to plug an Escherichia coli cell suspension first, to retain cells by positive DEP, and thereafter to recover them by washing with peptone water washing solution. This approach allows one not only to analyze DEP behavior of living cells within the chip, but also to further recover fractions containing DEP‐trapped cells. Bacterial concentration and flow rate appeared as critical parameters influencing the separation capacity of the chip. Evidence is presented demonstrating that the setup developed in this study can be used to separate different types of bacterial cells.

Dielectrophoresis of lambda-DNA using 3D carbon electrodes.

Carbon electrodes have recently been introduced as an alternative to metal electrodes and insulator structures for dielectrophoretic applications. Here, an experimental and theoretical study employing an array of 3D carbon electrodes contained in a microfluidic channel for the dielectrophoretic manipulation of DNA is presented. First evidence that carbon‐electrode DEP can be used for the manipulation and trapping of biomolecules such as DNA is reported. In particular, the dielectrophoretic response of λ‐DNA (48.5 kbp) under various frequencies and flow conditions necessary for retention of λ‐DNA are studied. Negative DEP is observed at frequencies above 75 kHz and positive DEP is present in the range below 75 kHz and down to 5 kHz. We further implement a theoretical model to capture the experimental findings in sufficient detail. Our theoretical considerations based on reported scaling laws for linear and supercoiled DNA further suggest that carbon‐electrode DEP devices could be employed in future analytical applications such as DNA preconcentration and fractionation.

SU-8 Photolithography as a Toolbox for Carbon MEMS

The use of SU-8 as precursor for glass-like carbon, or glassy carbon, is presented here. SU-8 carbonizes when subject to high temperature under inert atmosphere. Although epoxy-based precursors can be patterned in a variety of ways, photolithography is chosen due to its resolution and reproducibility. Here, a number of improvements to traditional photolithography are introduced to increase the versatility of the process. The shrinkage of SU-8 during carbonization is then detailed as one of the guidelines necessary to design carbon patterns. A couple of applications—(1) carbon-electrode dielectrophoresis for bioparticle manipulation; and (2) the use of carbon structures as micro-molds are also presented.

Very High Throughput Electrical Cell Lysis and Extraction of Intracellular Compounds Using 3D Carbon Electrodes in Lab-on-a-Chip Devices

Here we present an electrical lysis throughput of 600 microliters per minute at high cell density (108 yeast cells per ml) with 90% efficiency, thus improving the current common throughput of one microliter per minute. We also demonstrate the extraction of intracellular luciferase from mammalian cells with efficiency comparable to off-chip bulk chemical lysis. The goal of this work is to develop a sample preparation module that can act as a stand-alone device or be integrated to other functions already demonstrated in miniaturized devices, including sorting and analysis, towards a true lab-on-a-chip.

Increasing PCR sensitivity by removal of polymerase inhibitors in environmental samples by using dielectrophoresis

Dielectrophoresis (DEP) is a powerful tool to manipulate cells and molecules in microfluidic chips. However, few practical applications using DEP exist. An immediate practical application of a carbon-electrode DEP system, in removing PCR inhibitors from a sample, is reported in this work. We use a high throughput carbon-electrode DEP system to trap yeast cells from a natural sample (fermented grape must) and then in situ remove contaminants that interfere with PCR analysis. Retrieval of this enriched and purified yeast population from the DEP system then allows for a significant increase of sensitivity during PCR analysis. Furthermore, the fact that DEP can discriminate between viable and non-viable cells minimizes the number of false positives commonly obtained when using PCR alone. Experimental results provide clear evidence that the carbon-electrode DEP-based sample preparation step can readily and effectively clean environmental samples from natural contaminants and improve PCR sensitivity.

Enrichment of diluted cell populations from large sample volumes using 3D carbon-electrode dielectrophoresis

Here, we report on an enrichment protocol using carbon electrode dielectrophoresis to isolate and purify a targeted cell population from sample volumes up to 4 ml. We aim at trapping, washing, and recovering an enriched cell fraction that will facilitate downstream analysis. We used an increasingly diluted sample of yeast, 10(6)-10(2) cells/ml, to demonstrate the isolation and enrichment of few cells at increasing flow rates. A maximum average enrichment of 154.2 ± 23.7 times was achieved when the sample flow rate was 10 μl/min and yeast cells were suspended in low electrically conductive media that maximizes dielectrophoresis trapping. A COMSOL Multiphysics model allowed for the comparison between experimental and simulation results. Discussion is conducted on the discrepancies between such results and how the model can be further improved.

Perspectives of Micro and Nanofabrication of Carbon for Electrochemical and Microfluidic Applications

This chapter focuses on glass-like carbons, their method of micro and nanofabrication and their electrochemical and microfluidic applications. At first, the general properties of this material are exposed, followed by its advantages over other forms of carbon and over other materials. After an overview of the carbonization process of organic polymers we delve into the history of glass-like carbon. The bulk of the chapter deals with different fabrication tools and techniques to pattern polymers. It is shown that when it comes to carbon patterning, it is significantly easier and more convenient to shape an organic polymer and carbonize it than to machine carbon directly. Therefore the quality, dimensions and complexity of the final carbon part greatly depend on the polymer structure acting as a precursor. Current fabrication technologies allow for the patterning of polymers in a wide range of dimensions and with a great variety of tools. Even though several fabrication techniques could be employed such as casting, stamping or even Computer Numerical Controlled (CNC) machining, the focus of this chapter is on photolithography, given its precise control over the fabrication process and its reproducibility. Next Generation Lithography (NGL) tools are also covered as a viable way to achieve nanometer-sized carbon features. These tools include electron beam (e-beam), Focused-ion beam (FIB), Nano Imprint Lithography (NIL) and Step-and-Flash Imprint Lithography (SFIL). At last, the use of glass-like carbon in three applications, related to microfluidics and electrochemistry, is discussed: Dielectrophoresis, Electrochemical sensors, and Fuel Cells. It is exposed how in these applications glass-like carbon offers an advantage over other materials.

Challenges in the Use of Compact Disc-Based Centrifugal Microfluidics for Healthcare Diagnostics at the Extreme Point of Care

Since its inception, Compact Disc (CD)-based centrifugal microfluidic technology has drawn a great deal of interest within research communities due to its potential use in biomedical applications. The technology has been referred to by different names, including compact-disc microfluidics, lab-on-a-disk, lab-on-a-CD and bio-disk. This paper critically reviews the state-of-the-art in CD-based centrifugal microfluidics devices and attempts to identify the challenges that, if solved, would enable their use in the extreme point of care. Sample actuation, manufacturing, reagent storage and implementation, target multiplexing, bio-particle detection, required hardware and system disposal, and sustainability are the topics of focus.