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Publikationsliste Dr. Markus Rombach
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Originalarbeiten in wissenschaftlichen Fachzeitschriften Jahre: 2023 |
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2014 | alle anzeigen zurück zur Übersicht aller Publikationen G. Miotto, K. Thiemann, M. Rombach, R. Zengerle, S. KartmannHolographic PIV/PTV for nano flow rates - a study in the 70 to 200 nL/min range 2023 Biomedical engineering , Band : 68, Nummer : 1, Seiten : 97 - 107 G. Miotto, K. Thiemann, M. Rombach, R. Zengerle, S. KartmannHolographic PIV/PTV for nano flow rates–A study in the 70 to 200 nL/min range 2022 Biomedical Engineering S. Hin, N. Paust, M. Rombach, J. Lüddecke, M. Specht, R. Zengerle, K. MitsakakisMagnetophoresis in Centrifugal Microfluidics at Continuous Rotation for Nucleic Acid Extraction 2022 Micromachines , Band : 13, Seite : 2112 D. Baumgartner, B. Johannsen, M. Specht, J. Lüddecke, M. Rombach, S. Hin, N. Paust, F. von Stetten, R. Zengerle, C. Herz, J. R. Peham, P. N. Paqué, T. Attin, J. S. Jenzer, P. Körner, P. R. Schmidlin, T. Thurnheer, F. J. Wegehaupt, W. E. Kaman, A. Stubbs, J. P. Hays, V. Rusu, A. Michie, T. Binsl, D. Stejskal, M. Karpíšek, K. Bao, N. Bostanci, G. N. Belibasakis, K. MitsakakisOralDisk: A Chair-Side Compatible Molecular Platform Using Whole Saliva for Monitoring Oral Health at the Dental Practice 2021 Biosensors , Band : 11, Nummer : 11, Seite : 423 M. Rombach, S. Hin, M. Specht, B. Johannsen, J. Lüddecke, N. Paust, R. Zengerle, L. Roux, T. Sutcliffe, J. Peham, C. Herz, M. Panning, O. Donoso Mantke, K. MitsakakisRespiDisk: a point-of-care platform for fully automated detection of respiratory tract infection pathogens in clinical samples 2020 Analyst , Band : 145, Seiten : 7040 - 7047» Kurzfassung anzeigen « Kurzfassung verbergen Kurzfassung We present the RespiDisk enabling the fully automated and multiplex point-of-care (POC) detection of (currently) up to 19 respiratory tract infection (RTI) pathogens from a single sample based on reverse transcriptase polymerase chain reaction (RT-PCR). RespiDisk comprises a RTI-specific implementation of the centrifugal microfluidic LabDisk platform and combines new and existing advanced unit operations for liquid control, thereby automating all assay steps only by a spinning frequency and temperature protocol in combination with the use of a permanent magnet for in situ bead handing. The capabilities of the system were demonstrated with 36 tested quality samples mimicking clinical conditions (clinical and/or cultured material suspended in transport medium or synthetic bronchoalveolar lavage (BAL)) from past external quality assessment (EQA) panels covering 13 of the 19 integrated RTI detection assays. In total, 36 samples × 19 assays/sample resulting in 684 assays were performed with the RespiDisk, and its analytical performance was in full agreement with the routine clinical workflow serving as reference. A strong feature of the platform is its universality since its components allow the simultaneous detection of a broad panel of bacteria and viruses in a single run, thereby enabling the differentiation between antibiotic-treatable diseases. Furthermore, the full integration of all necessary biochemical components enables a reduction of the hands-on time from manual to automated sample-to-answer analysis to about 5 min. The study was performed on an air-heated LabDisk Player instrument with a time-to-result of 200 min. S. Hin, D. Baumgartner, M. Specht, J. Lüddecke, E. M. Arjmand, B. Johannsen, L. Schiedel, M. Rombach, N. Paust, F. von Stetten, R. Zengerle, N. Wipf, P. Müller, K. Mavridis, J. Vontas, K. Mitsakakis, * Indicates equally contributing authorsVectorDisk: A Microfluidic Platform Integrating Diagnostic Markers for Evidence-Based
Mosquito Control
2020 Processes , Band : 8, Nummer : 12, Seite : 1677» Kurzfassung anzeigen « Kurzfassung verbergen Kurzfassung Effective mosquito monitoring relies on the accurate identification and characterization of
the target population. Since this process requires specialist knowledge and equipment that is not
widely available, automated field-deployable systems are highly desirable. We present a centrifugal
microfluidic cartridge, the VectorDisk, which integrates TaqMan PCR assays in two feasibility studies,
aiming to assess multiplexing capability, specificity, and reproducibility in detecting disk-integrated
vector-related assays. In the first study, pools of 10 mosquitoes were used as samples. We tested
18 disks with 27 DNA and RNA assays each, using a combination of multiple microfluidic chambers
and detection wavelengths (geometric and color multiplexing) to identify mosquito and malaria
parasite species as well as insecticide resistance mechanisms. In the second study, purified nucleic
acids served as samples to test arboviral and malaria infective mosquito assays. Nine disks were tested
with 14 assays each. No false positive results were detected on any of the disks. The coeffcient of
variation in reproducibility tests was <10%. The modular nature of the platform, the easy adaptation
of the primer/probe panels, the cold chain independence, the rapid (2–3 h) analysis, and the assay
multiplexing capacity are key features, rendering the VectorDisk a potential candidate for automated
vector analysis. S. Hin, N. Paust, M. Keller, M. Rombach, O. Strohmeier, R. Zengerle, K. MitsakakisTemperature change rate actuated bubble mixing
for homogeneous rehydration of dry pre-stored
reagents in centrifugal microfluidics 2018 Lab Chip , Band : 18, Seiten : 362 - 370» Kurzfassung anzeigen « Kurzfassung verbergen Kurzfassung In centrifugal microfluidics, dead volumes in valves downstream of mixing chambers can hardly be avoided.
These dead volumes are excluded from mixing processes and hence cause a concentration gradient. Here
we present a new bubble mixing concept which avoids such dead volumes. The mixing concept employs
heating to create a temperature change rate (TCR) induced overpressure in the air volume downstream of
mixing chambers. The main feature is an air vent with a high fluidic resistance, representing a low pass filter
with respect to pressure changes. Fast temperature increase causes rapid pressure increase in downstream
structures pushing the liquid from downstream channels into the mixing chamber. As air further penetrates
into the mixing chamber, bubbles form, ascend due to buoyancy and mix the liquid. Slow temperature/
pressure changes equilibrate through the high fluidic resistance air vent enabling sequential heating/cooling
cycles to repeat the mixing process. After mixing, a complete transfer of the reaction volume into the
downstream fluidic structure is possible by a rapid cooling step triggering TCR actuated valving. The new
mixing concept is applied to rehydrate reagents for loop-mediated isothermal amplification (LAMP). After
mixing, the reaction mix is aliquoted into several reaction chambers for geometric multiplexing. As a measure
for mixing efficiency, the mean coefficient of variation (C
——
V, n = 4 LabDisks) of the time to positivity (tp)
of the LAMP reactions (n = 11 replicates per LabDisk) is taken. The C
——
V of the tp is reduced from 18.5%
(when using standard shake mode mixing) to 3.3% (when applying TCR actuated bubble mixing). The bubble
mixer has been implemented in a monolithic fashion without the need for any additional actuation besides
rotation and temperature control, which are needed anyhow for the assay workflow. S. Zehnle, M. Rombach, R. Zengerle, F. von Stetten, N. PaustNetwork simulation-based optimization of centrifugo-pneumatic blood plasma separation 2017 Biomicrofluidics , Band : 11, Seite : 024114» Kurzfassung anzeigen « Kurzfassung verbergen Kurzfassung Automated and robust separation of 14 ll of plasma from 40 ll of whole blood at a
purity of 99.81%60.11% within 43 s is demonstrated for the hematocrit range of
20%–60% in a centrifugal microfluidic polymer disk. At high rotational frequency,
red blood cells (RBCs) within whole blood are concentrated in a radial outer RBC
collection chamber. Simultaneously, plasma is concentrated in a radial inner
pneumatic chamber, where a defined air volume is enclosed and compressed.
Subsequent reduction of the rotational frequency to not lower than 25 Hz enables
rapid transfer of supernatant plasma into a plasma collection chamber, with highly
suppressed resuspension of red blood cells. Disk design and the rotational protocol
are optimized to make the process fast, robust, and insusceptible for undesired cell
resuspension. Numerical network simulation with lumped model elements is
used to predict and optimize the fluidic characteristics. Lysis of the remaining
red blood cells in the purified plasma, followed by measurement of the hemoglobin
concentration, was used to determine plasma purity. Due to the pneumatic
actuation, no surface treatment of the fluidic cartridge or any additional external
means are required, offering the possibility for low-cost mass fabrication technologies,
such as injection molding or thermoforming. M. Rombach, D. Kosse, B. Faltin, S. Wadle, G. Roth, R. Zengerle, F. von StettenReal-time stability testing of air-dried primers and
fluorogenic hydrolysis probes stabilized by trehalose
and xanthan 2014 Biotechniques , Band : 57, Nummer : 3, Seiten : 151 - 155» Kurzfassung anzeigen « Kurzfassung verbergen Kurzfassung A method for conserving primers and differently labeled fluorogenic hydrolysis (i.e., TaqMan) probes at ambient
conditions is presented. Primers and hydrolysis probes with four different fluorophore-quencher combinations (6-
FAM–BHQ1, HEX–BHQ1, ROX–BHQ650, and Cy5–BHQ2) were mixed with trehalose and xanthan at final concentrations
of 56 mM and 2.78 mM, respectively. Mixtures were air-dried at 23°C for 30 min on strips composed
of cyclo olefin polymer (COP), a material widely used in the manufacturing of in vitro diagnostic (IVD) test carriers.
After one year of storage, the functionality of the primers and fluorophore-quencher combinations was validated
by real-time polymerase chain reaction (real-time PCR), confirming their stability when stored in the presence of
stabilizers, with the best results achieved using trehalose. This approach could be of great benefit for manufacturing
IVD systems, for example, for genotyping applications based on multiplexing using different fluorescent dyes.
Konferenzbeiträge Jahre: 2020 |
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2009 | alle anzeigen zurück zur Übersicht aller Publikationen P. Koch, O. Barth, M. Meyer, R. Streller, R. Zengerle, M. Rombach, M. JehleMicrothermoforming: Enhancing blister technology to introduce microstructures in functional packaging 2020 12. European Thermoforming Conference, Geneva, Switzerland, 18.-20.03.2020 M. Rombach, S. Hin, M. Specht, B. Johannsen, J. Lüddecke, N. Paust, R. Zengerle, K. MitsakakisRespiDisk: A Point-of-Care platform for fully automated detection of respiratory tract infection pathogens in clinical samples 2020 MicroTAS 2020, 04.-09.10.2020, virtual 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. Hin, N. Paust, M. Rombach, J. Lueddecke, M. Specht, R. Zengerle, K. MitsakakisMinimizing ethanol carry-over in centrifugal microfluidic nucleic acid extraction by advanced bead handling and management of diffusive mass transfer 2019 Transducers 2019 - EUROSENSORS XXXIII, 23.-27. Juni 2019 - Berlin, Germany » Kurzfassung anzeigen « Kurzfassung verbergen Kurzfassung We present three concepts for centrifugal
microfluidics reducing ethanol carry-over in magnetic
bead-based nucleic acid (NA) extraction. Ethanol
carry-over is critical regarding inhibition of
downstream NA amplification. We identified two
possible carry-over pathways: Liquid co-transport
within bead-clusters and vapor diffusion. For the first
time, we integrated magnetic bead handling in
centrifugal microfluidics at continuous rotation aiming
to avoid liquid co-transport within bead-clusters.
Consequently, no significant contribution to ethanol
carry-over could be assigned anymore to liquid cotransport.
Major carry-over was attributed to diffusive
transport of ethanol vapor. Countermeasures reduced
this from 9.7 % (v/v) to 0.4 % (v/v), below the critical
level for inhibition of downstream amplification
reactions. 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. Rombach, S. Hin, O. Strohmeier, F. von Stetten, R. Zengerle, D. MarkPre-storage and release of purification reagents for full “hands-off” integration of DNA/RNA assays on the LabDisk platform 2014 MicroTAS 2014, San Antonio, USA, 26. – 30.10.2014 , Seiten : 1169 - 1171» Kurzfassung anzeigen « Kurzfassung verbergen Kurzfassung For the first time we demonstrate complete integration and pre-storage of all required compo-nents for nucleic acid extraction and purification on the centrifugal microfluidic LabDisk platform. On the one hand pre-storage of large volume liquid reagents from 200-–-550 μL is realized in stick-packs, that are stacked to decrease the footprint consumption on the LabDisk and are released au-tomatically during the run using centrifugal pressure (pcent-=-1 bar). On the other hand magnetic beads are mixed with PEG8000 (ratio 2:1) and air dried into the final cavity over 12 h. PEG8000 allows stable pre-storage and does not influence the extraction yield. S. Wadle, O. Strohmeier, M. Rombach, D. Mark, R. Zengerle, F. von StettenAutomatisierung der DNA Extraktion aus Vollblut unter Verwendung magnetischer Partikel auf der LabDisk Plattform 2013 Microsystemtechnik (MST) Kongress 2013, Aachen, 14. - 16.10.2013 , Seiten : 403 - 405» Kurzfassung anzeigen « Kurzfassung verbergen Kurzfassung Die Integration der Probenvorbereitung spielt eine Schlüsselrolle in der Entwicklung von point-of-care Diagnostik-
Systemen. Wir demonstrieren die Integration einer Magnetpartikel-basierten DNA Extraktion aus 200 μl Vollblut in eine
zentrifugal-mikrofluidische LabDisk. Die mithilfe der LabDisk extrahierte DNA (vier unabhängige Extraktionen aus
einer Blutprobe) wurde auf ihre Qualität geprüft. Als Referenz wurden dabei kommerziell verfügbare Säulen mit
Silicamembranen als Goldstandard der DNA-Extraktion verwendet. Es konnten gleich gute Ausbeuten einer
Wachstumsfaktor-Gensequenz – bestimmt mithilfe qPCR – (c1:10 LabDisk = 4,6 +/- 0,7 ng/μl gg. c1:10 Aufreinigungssäulen = 4,1
+/- 0,4 ng/μl) sowie Reinheiten (A260/A280 ~ 1,8 +/- 0,1) erzielt werden. Im Fall der LabDisk-extrahierten DNA lag
eine leicht erhöhte Ethanol-Konzentration (5,9 +/- 2,4) % im Vergleich zu (3,3 +/- 0,2) % bei Aufreinigungssäulen vor.
Die integierte und vollautomatiserte LabDisk-basierte DNA-Extraktion kann für weitere Anwendungen empfohlen
werden. O. Strohmeier, M. Rombach, D. Mark, R. Zengerle, G. Roth, F. von StettenErzeugung von Verdünnungsreihen auf einer Laborzentrifuge 2013 Microsystemtechnik (MST) Kongress 2013, Aachen, 14. - 16.10.2013 , Seiten : 353 - 356» Kurzfassung anzeigen « Kurzfassung verbergen Kurzfassung Vorgestellt wird ein neuartiges Verfahren zur automatischen Erzeugung von Verdünnungsreihen auf einer mikrofluidischen Kartusche. Die fluidische Prozessierung erfolgt lediglich durch definierte Rotation der Kartusche in einer Standard-Laborzentrifuge. Integrierte, aktiv gesteuerte Ventile sind hierbei nicht erforderlich. Das Verdünnungs-verhältnis wird lediglich durch das Volumen der zu verdünnenden Flüssigkeit bestimmt. Als Anwendungsbeispiel wird die Erzeugung von 5 Verdünnungsstufen aus Fluorescein in PBS Puffer in den Verhältnissen 1:3 und 1:5 gezeigt und anschließend die Reproduzierbarkeit mittels Fluoreszenzmessung gezeigt. S. Wadle, O. Strohmeier, M. Rombach, D. Mark, R. Zengerle, F. von StettenLabDisk integrated DNA Extraction from Whole Blood using Magnetic Particles 2012 MicroTAS 2012, Okinawa/Japan, 28.10. - 01.11.2012 , Seiten : 1381 - 1383 S. Zehnle, M. Rombach, F. von Stetten, R. Zengerle, N. PaustMicrofluidic centrifugo-pneumatic siphon enables fast blood plasma extraction with high yield and purity 2012 MicroTAS 2012, Okinawa/Japan, 28.10. - 01.11.2012 , Seiten : 869 - 871 M. Rombach, S. Lutz, C. Dumschat, A. Witt, S. Hensel, S. Frenzel, F. Aßmann, F. Gehring, T. Reiner, H. Drechsel, P. Szallies, D. Mark, G. Roth, R. Zengerle, F. von StettenSample-to-Answer LabDisk for Point-of-care Analysis of Total Cholesterol from Whole Blood 2012 MicroTAS 2012, Okinawa/Japan, 28.10. - 01.11.2012 , Seiten : 782 - 784 O. Strohmeier, M. Rombach, D. Mark, R. Zengerle, G. Roth, F. von StettenFully integrated dilution series generation on a laboratory centrifuge 2011 16th International Solid-State Sensors, Actuators and Microsystems Conference (TRANSDUCERS), 5-9 June 2011, Beijing China , Seiten : 2952 - 2955 » Kurzfassung anzeigen « Kurzfassung verbergen Kurzfassung We present a novel approach for the fully automated
generation of dilution series using a disposable microfluidic
cartridge. Fluids are metered, mixed and routed
without active valving by defined rotational speed only.
As a processing device, a standard lab centrifuge can be
used. The dilution ratio can be freely choosen by loading
the corresponding volume of sample into the cartridge
without the need for changes in the microfluidic layout.
Dilution series of Fluorescein in PBS buffer with ratios of
1:3 and 1:5 are shown with each dilution series
comprising 5 single dilution steps. Reproducibility was
determined by fluorescence measurement. D. Mark, M. Focke, S. Lutz, J. Burger, M. Müller, L. Riegger, M. Rombach, J. Hoffmann, G. Roth, O. Piepenburg, Y. Park, R. Zengerle, F. von StettenLab-on-a-chip solutions designed for being operated on standard laboratory instruments 2010 Eurosensors XXIV, September 5-8, Linz, Austria , Seiten : 444 - 447» Kurzfassung anzeigen « Kurzfassung verbergen Kurzfassung In this paper, we propose the development of microfluidic disposables that can be processed with standard laboratory
instruments. The use of prevalent processing devices could significantly reduce existing market entry barriers for lab-on-a-chip solutions and support the market uptake of microfluidic products. We demonstrate the concept with the following applications:
- microfluidic chips for DNA-purification operated on a standard laboratory centrifuge with 42% yield compared to gold
standards (QIAamp, Qiagen GmbH)
- microfluidic foil disk for DNA pre-amplification, aliquoting, and real-time PCR operated on a slightly modified Corbett Life
Science thermocycler (now Qiagen) with < 10 copy sensitivity
- microfluidic disposable for isothermal DNA amplification by recombinase polymerase amplification also operated on a
Corbett Life Science (now Qiagen) thermocycler with < 10 copy sensitivity and a time-to-result of < 15 minutes.
- fully automated hematocrit measurement in a DVD ROM drive from < 10 μL of whole blood. Martina Müller, Daniel Mark, Markus Rombach, Günter Roth, Jochen Hoffmann, Roland Zengerle, Felix von StettenOn the way to a fully integrated DNA-purification system on a standard laboratory centrifuge 2010 Proc. of the 14th International Conference on Miniaturized Systems for Chemistry and Life Sciences (MicroTAS), Groningen, The Netherlands, October 3 – 10 , Seiten : 405 - 408» Kurzfassung anzeigen « Kurzfassung verbergen Kurzfassung For the first time, we combine liquid reagent storage in glass capillaries [1], time-controlled reagent release [2], and a
novel solution for unidirectional centrifugal routing, paving the way to a fully integrated DNA extraction on a standard
laboratory centrifuge. The average yield of the extraction is 192 ± 30 ng DNA out of 32 μl blood, corresponding to
53 ± 8 % of a reference extraction. The overall processing time of ~66 minutes can be reduced to ~8 minutes after
optimization of the burst-valves. This novel approach demonstrates a convenient way for fully automated DNAextraction
on a standard laboratory centrifuge.
KEYWORDS: Microfluidics, centrifugal routing, DNA extraction, time-controlled release, liquid reagent storage Daniel Mark, Markus Rombach, Sascha Lutz, Roland Zengerle, Felix von StettenMicrofluidic unidirectional pneumatic switch for automated DNA-extraction on standard laboratory centrifuges 2009 Proc. of the Thirteenth International Conference on Miniaturized Systems for Chemistry and Life Sciences (µTAS), Jeju, Korea, November 1-5 , Seiten : 110 - 112» Kurzfassung anzeigen « Kurzfassung verbergen Kurzfassung For the first time, we present a new, unidirectional liquid switching concept for
the centrifugal microfluidic platform. The concept relies on a controlled liquid-air
interface instability. It enables automated DNA-extraction and potentially other microfluidic
protocols on standard laboratory centrifuges, superseding the need for expensive
base instruments, which are common to all state of the art Lab-on-a-Chip
platforms. As a proof of concept we fabricated a Lab-on-a-Chip cartridge for automated
DNA extraction from 32 μL whole blood on a standard laboratory centrifuge.
The obtained DNA yield was 88 ± 44 ng (42 % of the optimized reference extraction
[1]). Credits: SILK Icons by http://www.famfamfam.com/lab/icons/silk/