Biomedical subjects

Jörg P Kutter

Publications and source records attributed to Jörg P Kutter.

12 recordsLinked to original sources

PubMed2006

Towards a portable microchip system with integrated thermal control and polymer waveguides for real-time PCR.

A novel real-time PCR microchip platform with integrated thermal system and polymer waveguides has been developed. The integrated polymer optical system for real-time monitoring of PCR was fabricated in the same SU-8 layer as the PCR chamber, without additional masking steps. Two suitable DNA binding dyes, SYTOX Orange and TO-PRO-3, were selected and tested for the real-time PCR processes. As a model, cadF gene of Campylobacter jejuni has been amplified on the microchip. Using the integrated optical system of the real-time PCR microchip, the measured cycle threshold values of the real-time PCR performed with a dilution series of C. jejuni DNA template (2 to 200 pg/microL) could be quantitatively detected and compared with a conventional post-PCR analysis (DNA gel electrophoresis). The presented approach provided reliable real-time quantitative information of the PCR amplification of the targeted gene. With the integrated optical system, the reaction dynamics at any location inside the micro reaction chamber can easily be monitored.

Bacterial Outer Membrane Proteins↗

PubMed2006

Dielectrophoresis microsystem with integrated flow cytometers for on-line monitoring of sorting efficiency.

Dielectrophoresis (DEP) and flow cytometry are powerful technologies and widely applied in microfluidic systems for handling and measuring cells and particles. Here, we present a novel microchip with a DEP selective filter integrated with two microchip flow cytometers (FCs) for on-line monitoring of cell sorting processes. On the microchip, the DEP filter is integrated in a microfluidic channel network to sort yeast cells by positive DEP. The two FCs detection windows are set upstream and downstream of the DEP filter. When a cell passes through the detection windows, the light scattered by the cell is measured by integrated polymer optical elements (waveguide, lens, and fiber coupler). By comparing the cell counting rates measured by the two FCs, the collection efficiency of the DEP filter can be determined. The chips were used for quantitative determination of the effect of flow rate, applied voltage, conductivity of the sample, and frequency of the electric field on the sorting efficiency. A theoretical model for the capture efficiency was developed and a reasonable agreement with the experimental results observed. Viable and non-viable yeast cells showed different frequency dependencies and were sorted with high efficiency. At 2 MHz, more than 90% of the viable and less than 10% of the non-viable cells were captured on the DEP filter. The presented approach provides quantitative real-time data for sorting a large number of cells and will allow optimization of the conditions for, e.g., collecting cancer cells on a DEP filter while normal cells pass through the system. Furthermore, the microstructure is simple to fabricate and can easily be integrated with other microstructures for lab-on-a-chip applications.

Cell Separation↗

PubMed2005

AC electroosmotic pump with bubble-free palladium electrodes and rectifying polymer membrane valves.

We present the design, test and theoretical analysis of a novel micropump. The purpose is to make a pump with large flow rate (approximately 10 microL min-1) and high pressure capacity (approximately 1 bar) powered by a low voltage DeltaV<30 V. The pump is operated in AC mode with an electroosmotic actuator in connection with a full wave rectifying valve system. Individual valves are based on a flexible membrane with a slit. Bubble-free palladium electrodes are implemented in order to increase the range of applications and reduce maintenance.

Electrodes↗

PubMed2005

Long-term stable electroosmotic pump with ion exchange membranes.

We present the design, fabrication and test of a novel inline frit-based electroosmotic (EO) pump with ion exchange membranes. The pump is more stable than previous types due to a new flow component that ensures a controlled width of the diffusion layer close to the ion exchange membranes. The pump casing is constructed in polymers while the EO active part, the frit, is made in a nanoporous silica. The pressure capability of the pump is Deltapm/DeltaV = 0.15 bar V(-1). The flow rate to current ratio is Qm/I = 6 microL min(-1) mA(-1). This translates to Deltapm = 4.5 bar and Qm = 6 microL min(-1) at DeltaV = 30 V. The pump has been tested with four different buffer concentrations. In order to investigate day-to-day reproducibility each Q-p pump characteristic has been recorded several times during hour-long operation runs under realistic operating conditions.

Journal Article↗

PubMed2004

Effect of Joule heating on efficiency and performance for microchip-based and capillary-based electrophoretic separation systems: a closer look.

An attempt is made to revisit the main theoretical considerations concerning temperature effects ("Joule heating") in electro-driven separation systems, in particular lab-on-a-chip systems. Measurements of efficiencies in microfabricated devices under different Joule heating conditions are evaluated and compared to both theoretical models and measurements performed on conventional capillary systems. The widely accepted notion that planar microdevices are less susceptible to Joule heating effects is largely confirmed. The heat dissipation from a nonthermostatically controlled glass microdevice was found to be comparable to that from a liquid-cooled-fused silica capillary. Using typically dimensioned glass and glass/silicon microdevices, the experimental results indicate that 5-10 times higher electric field strengths can be applied than on conventional capillaries, before detrimental effects on the separation efficiency occur. The main influence of Joule heating on efficiency is via the establishment of a radial temperature profile across the lumen of the capillary or channel. An overall temperature increase of the buffer solution has only little influence on the quality of the separation. Still, active temperature control (cooling, thermostatting) can help prevent boiling of the buffer and increase the reproducibility of the results.

Electrophoresis↗

PubMed2004

Pure-silica optical waveguides, fiber couplers, and high-aspect ratio submicrometer channels for electrokinetic separation devices.

A new fabrication procedure for integration of ultraviolet transparent pure-silica planar waveguides, fiber couplers and high-aspect ratio submicrometer channels is presented. Only a single photolithographic mask step is required. The channels are 80-90 microm deep and the width can be reduced to about 0.5 microm, corresponding to a height-to-width ratio of more than 150. The core of the waveguides consists of pure silicon dioxide, which is favorable over doped silica, due to the absence of absorption centers associated with the dopants. This furthermore improves the long-term stability of the waveguides, because of an increased radiation resistance of the glass. The propagation loss decreases from 1.0 dB/cm at 200 nm to 0.2 dB/cm at 800 nm, which, to our knowledge, is the lowest propagation loss reported for integrated planar waveguides in the ultraviolet wavelength region to date. The effective optical path length is 1.2 mm for an absorbance cell with a nominal length of 1.0 mm, indicating effective suppression of stray light. The limit of detection for paracetamol when present in the entire channel network was determined to 3 microg/mL. Finally, the applicability of the fabricated devices for capillary electrophoresis was evaluated by separation of caffein, paracetamol and ketoprofone using absorbance detection at 254 nm.

Acetaminophen↗

PubMed2004

Recent developments in detection for microfluidic systems.

Microfluidic systems have become more and more important in the field of analytical chemistry. Detection methods on these microsystems are essential for the identification and quantification of chemical species that are being analyzed. This review concentrates on the latest developments of optical detection methods and mass spectrometry in conjunction with microfluidic systems. Electrochemical methods are discussed in another review in the same issue of this journal. Within the optical detection section, topics such as multiplexed detection and the use of waveguides are discussed. Within the discussion of mass spectrometry, the main focus is on electrospray emitters as interfaces between microsystem and spectrometer. Apart from optical detection and mass spectrometry, other techniques such as flame ionization and nuclear magnetic resonance are also mentioned.

Equipment Design↗

PubMed2003

A microfluidic device with an integrated waveguide beam splitter for velocity measurements of flowing particles by Fourier transformation.

A microfabricated capillary electrophoresis device for velocity measurements of flowing particles is presented. It consists of a 1 x 128 planar waveguide beam splitter monolithically integrated with an electrically insulated fluidic channel network for fluorescence excitation at multiple points. Stray light rejection structures are included in order to suppress unwanted light between the detection regions. The emission pattern of particles passing the detection region was collected by a photomultiplier tube that was placed in close proximity to the channel, thereby avoiding the use of transfer optics. The integrated planar waveguide beam splitter was, furthermore, permanently connected to the light source by a glued-on optical fiber, to achieve a robust and alignment-free operation of the system. The velocity was measured using a Fourier transformation with a Shah function, since the response of the light array was designed to approximate a square profile. Deviations from this response were observed as a result of the multimode nature of the integrated waveguides.

Journal Article↗

PubMed2003

CO2 laser microfabrication of an integrated polymer microfluidic manifold for the determination of phosphorus.

A simple colorimetric technique is implemented in a polymer microfluidic manifold. The simple chemistry aids an uncomplicated microchannel design, which is fabricated by CO(2) laser ablation. Issues such as bonding of multiple layers, alignment of micro-fabricated structures and integration of optical components are addressed. A demonstration of a stopped flow regime in the microfluidic manifold is also presented.

Journal Article↗

PubMed2003

Integration of polymer waveguides for optical detection in microfabricated chemical analysis systems.

Multimode polymer waveguides and fiber-to-waveguide couplers have been integrated with microfluidic channels by use of a single-mask-step procedure, which ensured self-alignment between the optics and the fluidics and allowed a fabrication and packaging time of only one day. Three fabrication procedures for obtaining hermetically sealed channels were investigated, and the spectrally resolved propagation loss (400-900 nm) of the integrated waveguides was determined for all three procedures. Two chemical absorbance cells with optical path lengths of 100 and 1000 microm were furthermore fabricated and characterized in terms of coupling loss, sensitivity, and limit of detection for measurements of the dye bromothymol blue.

Bromthymol Blue↗

PubMed2003

Separation and quantification of cellulases and hemicellulases by capillary electrophoresis.

Cellulases and hemicellulases are two classes of enzymes produced by filamentous fungi and secreted into the cultivation medium. Both classes of enzymes consist of a subset of classes of which the fungi produce several enzymes with varying molecular mass and pI but similar enzymatic activities. Current methods are limited in their ability to quantify all of these enzymes when all are present simultaneously in a mixture. Five different cellulases (two cellobiohydrolases and three endoglucanases) and one hemicellulase (endoxylanase) were separated using capillary electrophoresis (CE) in a fused silica capillary at pH values close to neutral. The improvement of the separation of these six proteins by the addition of alpha,omega-diaminoalkanes with chain lengths from three to seven carbon units was investigated. Dynamically coating the capillary with 1,3-diaminopropane resulted in separation of the six enzymes and the reproducibility of the migration times was between 0.6 and 1.9%. Two cases-quantitative determination of the enzyme concentrations in cultivation samples and investigation of adsorption of the enzymes onto cellulose-demonstrated the advantages and perspectives of CE analysis of these broad groups of enzymes.

Adsorption↗

PubMed2002

Performance of an in-plane detection cell with integrated waveguides for UV/Vis absorbance measurements on microfluidic separation devices.

A microfluidic device with integrated waveguides and a long path length detection cell for UV/Vis absorbance detection is presented. The 750 microm U-cell detection geometry was evaluated in terms of its optical performance as well as its influence on efficiency for electrophoretic separations in the microdevice. Stray light was found to have a strong effect on both, the sensitivity of the detection and the available linear range. The long path length U-cell showed a 9 times higher sensitivity when compared to a conventional capillary electrophoresis (CE) system with a 75 microm inner diameter (ID) capillary, and a 22 times higher sensitivity than with a 50 microm ID capillary. The linear range was comparable to that achieved in a 75 microm ID capillary and more than twice as large as in a 50 microm ID capillary. The use of the 750 microm U-cell did not contribute significantly to band broadening; however, a clear quantification was made difficult by the convolution of several other band broadening sources.

Absorption↗