Lukas Bogunovic

Electrodeless dielectrophoresis for bioanalysis: Theory, devices and applications

Dielectrophoresis is a non‐destructive, label‐free method to manipulate and separate (bio‐) particles and macromolecules. The mechanism is based on the movement of polarizable objects in an inhomogeneous electric field. Here, microfluidic devices are reviewed that generate those inhomogeneous electric fields with insulating posts or constrictions, an approach called electrodeless or insulator‐based dielectrophoresis. Possible advantages compared to electrode‐based designs are a less complex, monolithic fabrication process with low‐cost polymeric substrates and no metal surface deterioration within the area of sample analysis. The electrodeless design has led to novel devices, implementing the functionality directly into the channel geometry and covering many areas of bioanalysis, like manipulation and separation of particles, cells, DNA, and proteins.

Dielectrophoretic Trapping and Polarizability of DNA: The Role of Spatial Conformation

Dielectrophoresis is a convenient tool for controlled manipulation of DNA with numerous applications, including DNA trapping, stretching, and separation. However, the mechanisms behind the dielectrophoretic properties of DNA are still under debate, and the role of conformation has not been addressed yet. Here, we quantify dielectrophoretic effects on DNA by determining its polarizability from microfluidic single molecule trapping experiments. We systematically study different DNA configurations (linear and supercoiled, 6-164 kbp) and demonstrate that the polarizability strongly depends on the specific conformation and size of the DNA molecules. The connection to its spatial extension is established by measuring diffusion coefficients and from that the radii of gyration; details about the spatial DNA structure are obtained from atomic force microscopy images. For linear and supercoiled DNA fragments, we found a power-law scaling for the polarizabilities and the diffusion coefficients. Our results imply a scaling of the polarizability with the radius of gyration, alpha approximately Rg0.9+/-0.1 and alpha approximately Rg1.6+/-0.2 for linear and supercoiled DNA, respectively. As an application, we demonstrate the separation of DNA topoisomers based on their dielectrophoretic properties, achieving baseline resolution within 210 s. Purified DNA samples of specific configuration may be of great importance for DNA nanoassembly or future DNA vaccines.

Chiral Particle Separation by a Nonchiral Microlattice

We conceived a model experiment for a continuous separation strategy of chiral molecules (enantiomers) without the need of any chiral selector structure or derivatization agents: Microparticles that only differ by their chirality are shown to migrate along different directions when driven by a steady fluid flow through a square lattice of cylindrical posts. In accordance with our numerical predictions, the transport directions of the enantiomers depend very sensitively on the orientation of the lattice relative to the fluid flow.

Particle sorting by a structured microfluidic ratchet device with tunable selectivity: theory and experiment

We theoretically predict and experimentally demonstrate that several different particle species can be separated from each other by means of a ratchet device, consisting of periodically arranged triangular (ratchet) shaped obstacles. We propose an explicit algorithm for suitably tailoring the externally applied, time-dependent voltage protocol so that one or several, arbitrarily selected particle species are forced to migrate oppositely to all the remaining species. As an example we present numerical simulations for a mixture of five species, labelled according to their increasing size, so that species 2 and 4 simultaneously move in one direction and species 1, 3, and 5 in the other. The selection of species to be separated from the others can be changed at any time by simply adapting the voltage protocol. This general theoretical concept to utilize one device for many different sorting tasks is experimentally confirmed for a mixture of three colloidal particle species.

Photolithographic fabrication of arbitrarily shaped SU-8 microparticles without sacrificial release layers

We report on an efficient high throughput method for the photolithographic fabrication of well-defined arbitrarily shaped SU-8 microparticles without a sacrificial release layer. The procedure eliminates the spincoating of a sacrificial layer otherwise needed for particle lift-off, thereby reducing processing time and costs. Statistical analysis of the size distribution revealed a standard deviation of less than 2.3% in size. The particles can be immediately released into aqueous solution. This allows for anisotropical functionalization of the particles with, for example, biological loads or elements of molecular recognition after the development of the SU-8 structures.

Effective valve opening area in the detection of dysfunctional aortic valve prostheses: a differentiated statistical analysis of this parameter including the introduction of minimal expected normal values as borderline to dysfunctional stenotic prostheses.

Background: Dysfunction of heart valve prostheses (VP) is a life‐threatening complication and the diagnosis remains difficult. The motivation for this study was to improve the detection of dysfunctional VP by optimizing application of the prosthetic effective orifice area (VA). For this reason the minimal expected normal VA (VAexpected) was introduced. Methods: We investigated echocardiographically 1,369 normally functioning aortic valve prostheses (AVP). Mean VA, transprosthetic peak (PPG) and mean pressure gradients (MPG) were evaluated to gain reference values depending on prosthetic size and construction principle. Mean VAexpected was calculated by applying a simple formula that was developed empirically using statistical analyses. The results were compared with those of 65 dysfunctional AVPs. Results: VAexpected can be applied as a threshold between normal and dysfunctional stenotic AVP and showed a correct estimation in 87% of all normally functioning and 100% of dysfunctional stenotic VPs. The sensitivity for all prosthetic sizes is 1.0, independently of the constructional principle of the VP. Specificity ranged between 0.8 and 1.0, dependent on VP size. The formula representing VAexpected is simple and can be executed easily. Conclusion: As nearly independent of stroke volume and in consideration of VAexpected, VA seems to have become one of the preferable parameters for detecting pathological stenotic AVPs echocardiographically. The additional application of PPG/MPG and other parameters permits prostheses with relevant isolated regurgitation and patient‐prosthesis‐mismatch to be distinguished. (Echocardiography 2012;29:713‐719)

Fabrication of a microfluidic channel with differently modified surfaces with a two component approach

We report on a two-component fabrication technique for microfluidic channels, allowing for different chemical or physical surface modifications of channel walls within one single channel. The two components are made of polydimethylsiloxane and prepared via soft lithography independently. After appropriate pre-treatment with the desired functions, the two parts are bonded together using oxygen plasma and a Fineplacer® lambda system. As a proof of concept, we present the combination of electroosmosis and opposing hydrodynamic flow in a microfluidic channel leading to different velocity presigns of the resulting flow on opposite channel walls, due to different surface modifications. These results indicate an intact Pluronic® F108 surface coating after assembly.