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Publikationsliste Dr. Marc Karle
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Originalarbeiten in wissenschaftlichen Fachzeitschriften Jahre: 2011 |
2010 | alle anzeigen zurück zur Übersicht aller Publikationen Marc Karle, Johannes Wöhrle, Junichi Miwa, Nils Paust, Günter Roth, Roland Zengerle, Felix von StettenControlled counter-flow motion of magnetic bead chains rolling along microchannels 2011 Microfluid Nanofluid , Band : 10, Nummer : 4, Seiten : 935 - 939» Kurzfassung anzeigen « Kurzfassung verbergen Kurzfassung We demonstrate controlled transport of superparamagnetic
beads in the opposite direction of a laminar
flow. A permanent magnet assembles 200 nm magnetic
particles into about 200 lm long bead chains that are
aligned in parallel to the magnetic field lines. Due to a
magnetic field gradient, the bead chains are attracted
towards the wall of a microfluidic channel. A rotation of
the permanent magnet results in a rotation of the bead
chains in the opposite direction to the magnet. Due to
friction on the surface, the bead chains roll along the
channel wall, even in counter-flow direction, up to at a
maximum counter-flow velocity of 8 mm s(-1). Based on
this approach, magnetic beads can be accurately manoeuvred
within microfluidic channels. This counter-flow
motion can be efficiently be used in Lab-on-a-Chip systems,
e.g. for implementing washing steps in DNA
purification. Marc Karle, Junichi Miwa, Gregor Czilwik, Volker Auwärter, Günter Roth, Roland Zengerle, Felix von StettenContinuous microfluidic DNA extraction using phase-transfer magnetophoresis 2010 Lab Chip , Band : 10, Seiten : 3284 - 3290» Kurzfassung anzeigen « Kurzfassung verbergen Kurzfassung This paper reports a novel microfluidic-chip based platform using "phase-transfer magnetophoresis"
enabling continuous biomolecule processing. As an example we demonstrate for the first time
continuous DNA extraction from cell lysate on a microfluidic chip. After mixing bacterial Escherichia
coli culture with superparamagnetic bead suspension, lysis and binding buffers, DNA is released from
cells and captured by the beads. These DNA carrying beads are continuously transported across the
interfaces between co-flowing laminar streams of sample mixture, washing and elution buffer. Bead
actuation is achieved by applying a time-varying magnetic field generated by a rotating permanent
magnet. Flagella-like chains of magnetic beads are formed and transported along the microfluidic
channels by an interplay of fluid drag and periodic magnetic entrapment. The turnover time for DNA
extraction was approximately 2 minutes with a sample flow rate of 0.75 µl/s and an eluate flow rate of
0.35 µl/s. DNA recovery was 147% (on average) compared to bead based batch-wise extraction in
reference tubes within a dilution series experiment over 7 orders of magnitude. The novel platform is
suggested for automation of various magnetic bead based applications that require continuous sample
processing, e.g. continuous DNA extraction for flow-through PCR, capture and analysis of cells and
continuous immunoassays. Potential applications are seen in the field of biological safety monitoring,
bioprocess control, environmental monitoring, or epidemiological studies such as monitoring the load
of antibiotic resistant bacteria in waste water from hospitals.
Reviews/Übersichtsartikel in wissenschaftlichen Fachzeitschriften Jahre: 2016 | alle anzeigen zurück zur Übersicht aller Publikationen M. Karle, S. K. Vashist, R. Zengerle, F. von StettenMicrofluidic solutions enabling continuous processing and
monitoring of biological samples: A Review 2016 Anal Chim Acta , Band : 929, Seiten : 1 - 22» Kurzfassung anzeigen « Kurzfassung verbergen Kurzfassung The last decade has witnessed tremendous advances in employing microfluidic solutions enabling Continuous Processing and Monitoring of Biological Samples (CPMBS), which is an essential requirement for the control of bio-processes. The microfluidic systems are superior to the traditional inline sensors due to their ability to implement complex analytical procedures, such as multi-step sample preparation, and enabling the online measurement of parameters. This manuscript provides a backgound review of microfluidic approaches employing laminar flow, hydrodynamic separation, acoustophoresis, electrophoresis, dielectrophoresis, magnetophoresis and segmented flow for the continuous processing and monitoring of biological samples. The principles, advantages and limitations of each microfluidic approach are described along with its potential applications. The challenges in the field and the future directions are also provided.
Kurzbeiträge Jahre: 2018 | alle anzeigen zurück zur Übersicht aller Publikationen J. Schemberg, M. Specht, M. KarleNeue Wege in der Krebsdiagnostik - Mikrofluidische Strategien für die Medizin 4.0 2018 GIT-Laborfachzeitschrift , Band : 4, Seite : 16
Konferenzbeiträge Jahre: 2019 |
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2008 | alle anzeigen zurück zur Übersicht aller Publikationen J. Weisemann, G. B. Stevens, T. van Oordt, N. Krez, K. Dormanns, M. Karle, F. von Stetten, A. RummelFast automatic detection of Botulinum neurotoxin by the BoNT LabDisk assay: results of a trans-european performance study 2019 Toxins 2019, Copenhagen/Denmark, 16. - 19.01.2019 » Kurzfassung anzeigen « Kurzfassung verbergen Kurzfassung Botulinum Neurotoxins (BoNT) are the most toxic naturally occurring substances for humans. After potential intoxication and occurrence of botulism symptoms, urgent detection of causative BoNT is highly desirable for successful medical treatment. The mouse bioassay (MBA) as gold standard requires a large number of mice, takes several days and is not suitable for point-of-care.
Here, we developed an automatic in vitro assay to detect BoNT in complex biological matrices. The assay detects specific cleavage of SNARE proteins by BoNTs by fluorescent read out in a centrifugal microfluidic cartridge, termed BoNT LabDisk (BLD). The generic LabDisk comprises two different peptides able to distinguish between BoNT/ACE and BDF, respectively. The BLD prototype system was tested in a trans-European performance study by five naïve laboratories with expertise in BoNT detection to demonstrate practical applicability and performance of the BLD system. Eight blinded samples containing three complex biological matrices (human serum, bean juice, carrot juice) or buffer spiked with 10-1000 pM of BoNT/A and D were provided. Stability of samples was confirmed by mouse hemidiaphragm assay prior to shipment.
The laboratories detected absence or presence of BoNT/A in 34 of the 40 samples correctly (85% accuracy). In three of ten bean juice or serum samples problems occurred due to high viscosity which requires improvement of sample preparation. In another three samples, signals close to background led to misinterpretation by experimentators. Altogether, the performance study showed that handling of the BLD assay is very easy, fast and practicable. The current detection limit is close to that of MBA.
In conclusion, the BLD system allows the detection of functionally active BoNT in short assay times with low limits of detection within different matrices, without the need of animals or animal based reagents. The next step will be the adaption of the BLD assay for a direct serotyping of BoNT/A-F. M. Specht, J. Schemberg, T. Förster, S. Burger, M. Rombach, N. Paust, R. Zengerle, F. von Stetten, G. Gastrock, M. KarleMicrofluidic App for centrifugal separation and purification of lymphatic cancer cells from whole blood 2019 MST-Kongress, 28. - 30.Oktober 2019, Berlin S. Burger, J. Schemberg, T. Förster, M. Specht, M. Rombach, N. Paust, R. Zengerle, M. KarleSeparation of low-abundance cells as an App on standard laboratory centrifuge 2017 MicroTAS 2017, Savannah /USA, 22.-26.10.2017 » Kurzfassung anzeigen « Kurzfassung verbergen Kurzfassung A novel centrifugal microfluidic cell separation module is presented to extract target cells (KG-1) from buffy coat using antibody coated microbeads. All structures required for the target cell separation are monolithically integrated in the microfluidic cartridge including mixing under constant rotation using oxygen bubbles. Thus, the microfluidic cartridge processing can be done on a standard laboratory centrifuge lowering the monetary investment for the user. The entire process chain of the module is completed within 26 minutes and recovers approximately 90 % of the lympathic cells in the sample while only requiring few manual pipetting steps. M. Rombach, M. Keller, N. Paust, F. von Stetten, D. Mark, R. Zengerle, M. KarleThe LabCard – A new approach for centrifugal assay automation 2017 MST Kongress, München, 23. - 25.10.2017 » Kurzfassung anzeigen « Kurzfassung verbergen Kurzfassung We present the LabCard, a new centrifugal microfluidic approach for simultaneous processing of multiple test carriers by arranging them in the vertical plane. As a proof-of-concept, we fluidically integrated an assay for isothermal nucleic acid analyses of respiratory pathogens with the microfluidic network comprising unit operations and processing chains[1] for thermal lysis of a patient sample (25–75 μL), release of pre-stored reagents, mixing of reagents, aliquoting and distribution into amplification wells, which was successfully demonstrated in 5/5 runs with an overall fluidic processing time of 10 min. M. Rombach, M. Keller, N. Paust, F. von Stetten, D. Mark, R. Zengerle, M. KarleThe LabCard – A new approach for centrifugal assay automation” 2016 20th International Conference on Miniaturized Systems for Chemistry and Life Sciences, µTAS 2016, Dublin / Irland, 09. – 13.10.2016 M. Rombach, S. Zehnle, N. Paust, M. Weil, Ö. Sogukpinar, R. Zengerle, M. KarleMicrofluidic App for buffy coat extraction from large peripheral blood samples for low-abundance living-cell analysis 2015 19th International Conference on Miniaturized Systems for Chemistry and Life Sciences October 25-29, 2015, Gyeongju, KOREA M. Karle, J. Wöhrle, F. von Stetten, R. Zengerle, D. MarkAxial centrifugal filtration – A novel approach for rapid bacterial concentration from a large volume 2013 Transducers 2013, Barcelona, Spain, 16. – 18.06.2013 , Seiten : 1235 - 1238» Kurzfassung anzeigen « Kurzfassung verbergen Kurzfassung A novel approach for filtration on a centrifugal microfluidic platform is presented for the first time. This approach is intended to concentrate bacteria from a large volume. In axial centrifugal filtration the filter is oriented perpendicular to the axis of rotation. This feature allows for integration of dead-end filtration while the filter cake is continuously removed from the filter by centrifugation. This prevents clogging of the filter. Furthermore, a continuous sample feed enables processing of large samples on one disk. Especially for analyzing drinking water large volumes have to be processed and solutions for rapid bacterial concentration are highly appreciated. M. Karle, G. Czilwik, J. Miwa, N. Paust, G. Roth, R. Zengerle, F. von StettenContinuous microfluidic DNA purification for online monitoring and process control 2011 Proceedings of 2011 International Conference on Microtechnologies in Medicine and Biology Lucerne, Switzerland, 4-6 May , Seiten : 197 - 198» Kurzfassung anzeigen « Kurzfassung verbergen Kurzfassung We demonstrate the first continuous microfluidic platform for flow-through DNA purification from cell lysate. Using superparamagnetic beads, DNA is continuously transported across interfaces between co-flowing laminar streams of extraction reagents. In on-chip experiments DNA was continuously purified over a time period of 110 min. After the on-chip purification, DNA content and purity of the eluate sampled at different stages of the continuous extraction experiment was analyzed off-chip via qPCR. Results show successful flow-through purification with constant output over almost the complete duration of the experiment. Possible applications are seen in biological safety and environmental monitoring or bioprocess control. M. Karle, G. Czilwik, J. Miwa, N. Paust, G. Roth, R. Zengerle, F. von StettenMicrofluidic flow-through DNA purification for continuous monitoring applications 2011 EuroMedLab Berlin 2011 – Berlin, 15-19 May 2011 Clin Chem Lab Med , Band : 49, Ergänzungsband : 1, Seiten : S607 - S607 M. Karle, G. Czilwik, J. Miwa, N. Paust, G. Roth, R. Zengerle, F. von Stetten, Felix von StettenHigh-performance flow-through DNA purification on a microfluidic chip 2010 Proc. of the 14th International Conference on Miniaturized Systems for Chemistry and Life Sciences (MicroTAS), Groningen, The Netherlands, October 3 – 10 , Seiten : 106 - 108» Kurzfassung anzeigen « Kurzfassung verbergen Kurzfassung We present a significant increase in performance for flow-through purification of nucleic acids by continuous
microfluidic processing. After binding to superparamagnetic beads the nucleic acids are sequentially transported across
the phase-interface of co-flowing laminar streams of purification reagents. The entire purification procedure is
performed within only 2 minutes. Compared to classical batch-wise purification in test tubes, 150 50 % of total DNA
have been recovered in on-chip purifications over a DNA concentration range of 7 orders of magnitude. With
appropriate surface modification of the magnetic beads the chip is also suggested for the implementation of other
continuous biomolecular purification tasks.
KEYWORDS: Nucleic Acids Purification, Magnetophoresis, Continuous Processing, Flow-Through Assay M. Karle, J. Miwa, G. Roth, R. Zengerle, F. von StettenA novel microfluidic platform for continuous DNA extraction and purification using laminar flow magnetophoresis 2009 Proc. IEEE-MEMS; 25 – 29 January 2009, Sorrento, Italy , Seiten : 276 - 279» Kurzfassung anzeigen « Kurzfassung verbergen Kurzfassung For the first time we present a novel microfluidic platform using laminar-flow magnetophoresis for combined continuous extraction and purification of DNA from cells. All essential unit operations (DNA binding, sample washing and DNA elution) are integrated on one single chip. The key function is the motion of magnetic beads given by the interplay of laminar flow and a time-varying magnetic field. The magnetic beads enable the transport of the DNA across the interfaces between co-flowing laminar streams in a circular channel arrangement around a central rotating permanent magnet inducing time-varying magnetic field, which prevents the beads from sticking to the channel walls and enables controlled transfer of beads between different extraction reagents. The system was fabricated by micromilling in polycarbonate. Pressure driven experiments were performed to analyze the magnetophoretic concept and the biotechnical functionality. Compared to a macroscopic reference system 25% of total DNA have been recovered. An inlet flow velocity of 12.5 mm∙s-1 lead to an average bead velocity of 1.6 mm∙s-1 The sample transition time is approximately 1 minute. The device is a central part of a complete biochemical sensing system for continuous monitoring of cell growth in bioreactors, but allows also continuous purification of DNA, RNA, proteins or cells, including their subsequent real-time analysis. Marc Karle, Junichi Miwa, Günter Roth, Stefan Haeberle, Roland Zengerle, Felix von StettenKontinuierlich arbeitende Mikrofluidik-Plattform zur Aufreinigung von Biomolekülen 2009 Tagungsband Mikrosystemtechnik-Kongress, Berlin, 12.-14. Oktober , Seite : P 31» Kurzfassung anzeigen « Kurzfassung verbergen Kurzfassung Wir präsentieren eine neue Mikrofluidik-Plattform, die eine kontinuierliche Extraktion und Aufreinigung von Biomolekülen z.B. aus einem Fermenter ermöglicht. Die Plattform beruht auf dem Einsatz der Magnetophorese in einer Laminarströmung. Alle erforderlichen Arbeitsschritte (Binden, Waschen und Eluieren der Biomoleküle) sind auf einem Chip integriert. Eine Schlüsselfunktion nehmen superparamagnetische Partikel in einer Laminarströmung in Verbindung mit einem zeitlich variierenden Magnetfeld ein. Die Partikel ermöglichen den Transport der Biomoleküle über die Phasengrenze der Laminarströmung in einer kreisförmig um einen zentralen, rotierenden Permanentmagneten angeordneten Kanalstruktur. Die Rotation des Permanentmagneten erzeugt ein periodisch variierendes Magnetfeld, das die kontrollierte Bewegung der magnetischen Partikel zwischen unterschiedlichen Extraktionsreagenzien ermöglicht, ohne die Partikel an der Kanalwand zu immobilisieren. Als erste Applikation wurde ein Chip zur DNA-Extraktion aus Polykarbonat mittels Mikrofräsen hergestellt. Um die biotechnische Funktionalität und das magnetophoretische Konzept zu untersuchen, wurde bakterielle DNA direkt aus dem Lysat extrahiert. Im Vergleich mit einem makroskopischen Referenzsystem wurden 80 % der DNA aufgereinigt. Eine Eingangsflussgeschwindigkeit von 12.5 mm∙s-1 führte zu einer durchschnittlichen Partikelgeschwindigkeit von 1.6 mm∙s-1 und zu einer DNA-Extraktion innerhalb ca. 1 Minute. Die Plattform ist ein zentrales Element eines Überwachungssystems zur kontinuierlichen Kontrolle des Zellwachstums in Bioreaktoren. Darüber hinaus kann sie, je nach Oberflächenmodifikation der magnetischen Partikel, auch für die Aufreinigung von RNA, Proteinen oder Zellen für nachfolgende Echtzeitanalysen verwendet werden. T Metz, D Mark, S Lutz, O Strohmeier, D Kosse, M Focke, B Faltin, J Burger, J Böning, J Miwa, M Karle, G Müller, C Müller, S Messner, H Reinecke, R Zengerle, F von StettenLab-on-a-Chip Foundry Service – enabling the miniaturization of in vitro diagnostics 2009 Euromedlab, Innsbruck, Austria, 2009, 350-368 Clinical Chemistry and Laboratory Medicine , Seiten : 350 - 368 T. Metz, M. Karle, T. Preis, D. Mark, G. Roth, C. Müller, R. Zengerle, F. von StettenLab-on-a-Chip Foundry Service: Schnelle kundenspezifische Implementierung miniaturisierter biochemischer Assays 2009 Tagungsband Mikrosystemtechnik-Kongress 2009, Berlin, 12.-14. Oktober , Seite : P 42» Kurzfassung anzeigen « Kurzfassung verbergen Kurzfassung Wir präsentieren eine konsistente Methodik zur effizienten Miniaturisierung, Integration und Automatisierung bio-chemischer Assays auf der Grundlage definierter mikrofluidischer Plattformen [1]. Eine mikrofluidische Plattform stellt einen Satz fluidischer Grundfunktionen (sogenannte Einheitsoperationen) zur Verfügung welche einfach kombiniert und durch eine günstige, wohl definierte Fertigungstechnologie hergestellt werden können. Anwendungsspezifische Assays lassen sich hierüber flexibel und mit geringem Entwicklungsrisiko durch die systematische Verknüpfung plattformspezifischer, validierter, Einheitsoperationen implementieren. Die mikrofluidischen Layouts der Einheitsoperationen werden in einem Design Handbuch, zusammen mit Validierungsergebnissen und den Fertigungsprozessen dokumentiert. Für die Validierung werden standardisierte Strukturen eingesetzt. Dies wird beispielhaft anhand einer Teststruktur für die kapillare Befüllung gezeigt. Daniel Mark, Stefan Haeberle, Junichi Miwa, Patrick Weber, Guenter Roth, Marc Karle, Max Focke, Sascha Lutz, Jochen Hoffmann, Claas Müller, Felix von Stetten, Roland ZengerleTwo Microfluidic Platforms for Miniaturization, Integration and Automation of Assays 2009 Proc. of COMS, Copenhagen , Seite : -» Kurzfassung anzeigen « Kurzfassung verbergen Kurzfassung Two different platform concepts for microfluidic miniaturization, integration and automation of biochemical assays are presented. First, unit operations for batch-wise nucleic acid analysis on the centrifugal microfluidic platform are demonstrated, including unit operations for DNA extraction, aliquoting and real-time PCR. Second, the newly developed continuous phase transfer magnetophoresis platform is introduced. It enables continuous online process monitoring, demonstrated by implementation of unit operations for DNA extraction. J. Böning, D. Mark, S. Lutz, B. Faltin, M. Focke, M. Karle, J. Ducrée, S. Messner, R. Zengerle, F. von Stetten"Lab-on-a-Chip Foundry Service": A Systematic Approach to the Development of Centrifugal Microfluidic Technologies 2008 Actuator 2008 / Bremen Messe Bremen, Seiten : 814 - 817» Kurzfassung anzeigen « Kurzfassung verbergen Kurzfassung This contribution provides a novel approach to a time and cost-efficient development of polymeric lab-on-a-chip
applications. Rather than starting from scratch, new designs are derived from a library of validated laboratory
unit operations (LUOs) and prototyped by standard operating procedures (SOPs). LUOs as well as SOPs are
systematically documented and retrieved from a "BlueBook".
The essential ingredients for the development of lab-on-a-chip applications are liquid handling technologies and
fabrication technologies as well as test and development tools. This paper outlines a streamlined approach based
on a microfluidic platform concept to reduce the cost, time and risks for the development of lab-on-a-chip
technologies. As an example illustrating the platform concept, we describe our centrifugal microfluidic “Lab-ona-
Disk” platform.
The paradigm of the platform concept is to establish a library of LUOs such as metering, mixing and routing
which are validated by experiments and accompanying simulations for a certain parameter range. The
concatenation of these LUOs allows to realize complex applications. The layout is then transferred into
hardware according to SOPs and afterwards measured at a designated test and development stand. Design rules
for the LUOs, procedures (SOPs) and other relevant information such as materials or device components are
documented and retrieved from a wiki-based software-based knowledge management portal called “BlueBook”.
Keywords: Lab-on-a-Chip Foundry Service, centrifugal microfluidic platform, standard operating procedures,
laboratory unit operations, knowledge management system T Metz, D Mark, S Lutz, R Simon, M Focke, B Faltin, J Burger, J Böning, J Miwa, M Karle, G Müller, C Blattert, C Müller, S Messner, H Reinecke, R Zengerle, F von StettenLab-on-a-Chip Foundry Service – speeding up the miniaturization of in vitro diagnostics 2008 Proc. 3rd Technology Forum Diagnostics & Bioanalytical Devices, Frankfurt, 2008, 3 , Band : 3» Kurzfassung anzeigen « Kurzfassung verbergen Kurzfassung We present the Lab-on-a-Chip Foundry Service of the HSG-IMIT that was founded to
speed up the realization of in vitro diagnostic assays (IVD) as miniaturized
microfluidic systems (Lab-on-a-Chip). The Lab-on-a-Chip Foundry Service opens the
advantages of Lab-on-a-Chip - smaller consumption of reagents and faster
turnaround times in compact, automated diagnostics - to a broad community. We
offer everything from one hand: Layout, prototyping, fluidic and biochemical testing. Credits: SILK Icons by http://www.famfamfam.com/lab/icons/silk/