The LabTube – a novel microfluidic platform for assay automation in laboratory centrifuges

Kloke, Arne; Fiebach, A. R.; Zhang, S.; Drechsel, Lisa; Niekrawietz, S.; Hoehl, Melanie M.; Kneusel, Richard E.; Panthel, K.; Steigert, J.; Stetten, Felix von; Zengerle, Roland; Paust, Nils

doi:10.1039/c3lc51261d

Cited by 30 publications

(43 citation statements)

References 29 publications

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“…Here, pen mechanics, actuated by centrifugal forces, stepwise rotate Revolver II by changing the centrifugal acceleration, which affords opening and closing of fluidic paths through the stack (Kloke et al, 2014). The mechanics of the modified LabT ube system was successfully verified as Revolver II moved from the start (S) to end (E) position during the centrifugation protocol, whilst all components remained intact (Fig.…”

Section: Mechanic Fluidic and Electric Verification During Operationmentioning

confidence: 97%

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Centrifugal LabTube platform for fully automated DNA purification and LAMP amplification based on an integrated, low-cost heating system

Hoehl

Weißert

Dannenberg

et al. 2014

Biomed Microdevices

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Abstract:This paper introduces a disposable battery-driven heating system for loop-mediated isothermal DNA amplification (LAMP) inside a centrifugally-driven DNA p urification platform (LabT ube). We demonstrate LabT ube-based fully automated DNA purification of as low as 100 cell-equivalents of verotoxin-producing Escherichia coli (VT EC) in water, milk and apple juice in a laboratory centrifuge, followed by integrated and automated LAMP amplification with a reduction of hands-on time from 45 to 1min. The heating system consists of two parallel SMD thick film resistors and a NT C as heating and temperature sensing elements. They are driven by a 3V battery and controlled by a microcontroller. The LAMP reagents are stored in the elution chamber and the amplification starts immediately after the eluate is purged into the chamber. The LabT ube, including a microcontroller-based heating system, demonstrates contamination-free and automated sample-to-answer nucleic acid testing within a laboratory centrifuge. The heating system can be easily parallelized within one LabT ube and it is deployable for a variety of heating and electrical applications.

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Section: Mechanic Fluidic and Electric Verification During Operationmentioning

confidence: 97%

“…The LabT ube is based on disposable, polymeric modules integrated in a 50ml falcon tube (Kloke et al, 2014), in which the DNA purification workflow is automated by applying processspecific centrifugation-time-protocols to a centrifuge (Fig. 1).…”

Section: Introductionmentioning

confidence: 99%

Centrifugal LabTube platform for fully automated DNA purification and LAMP amplification based on an integrated, low-cost heating system

Hoehl

Weißert

Dannenberg

et al. 2014

Biomed Microdevices

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show abstract

“…The operation time of this device is reported as~1.5 h to fractionate 60-200 kbp DNA 13,14 . Another reported variant of the 'DNA prism' device was a self-patterned 3D crystalline nanoarray used for fractionating smaller DNA molecules (2-50 kbp) in a continuous flow setup 15 .…”

Section: Introductionmentioning

confidence: 99%

Exploiting biased reptation for continuous flow preparative DNA fractionation in a versatile microfluidic platform

et al. 2017

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A new approach is presented for preparative, continuous flow fractionation of sub-10-kbp DNA fragments, which exploits the variation in the field-dependent mobility of the DNA molecules based on their length. Orthogonally pulsed electric fields of significantly different magnitudes are applied to a microchip filled with a sieving matrix of 1.2% agarose gel. Using this method, we demonstrate a high-resolution separation of 0.5, 1, 2, 5, and 10 kbp DNA fragments within 2 min. During the separation, DNA fragments are also purified from other ionic species. Preparative fractionation of sub-10-kbp DNA molecules plays an important role in second-generation sequencing. The presented device performs rapid high-resolution fractionation and it can be reliably manufactured with simple microfabrication procedures. This method has the advantages of simplicity, versatility, and reproducibility. However, it suffers from long processing times on the order of a few tens of hours 2,3 . For instance, the Bio-Rad CHEF-DR device performs fractionation of 5-120 kbp DNA using pulsed-field gel electrophoresis (PFGE) in 25 h (Refs. 4,5). Trends toward second-generation sequencing motivate the replacement of standard gel electrophoresis with microchip-based systems, which could provide efficient platforms to minimize the processing time and to perform optimal DNA fractionation 6-8 .Micro-and nanofabricated post arrays-that resemble a gel matrix with well-ordered and identical pores-have been integrated in microchip-based systems, increasing both the understanding of DNA separation principles and the fractionation's efficiency and speed. Two decades ago, Volkmuth and Austin 9 introduced a patterned micro-post array in a microfluidic electrophoresis platform, in which DNA fragments were separated by biased reptation under a DC electric field. Later, Duke et al. separated large fragments in a range of 60-135 kbp in a microfabricated array using DNA reorientation-or the 'switchback' principle. Although the switchback principle is similar to what is used in PFGE, the device yields much faster separations owing to its sparse and regular sieving array 10 . In a later work, Kaji et al. studied a three-dimensional (3D) nanopost array as an optimal separation matrix for DNA fragments over a few kbps under a DC electric field, resulting in similarly fast separations due to the sparse matrix 11 .With the aim of increasing the sample throughput and further facilitating the sample recovery, a continuous flow separation was developed. Huang et al. 12 performed continuous flow PFGE in a micromachined post array by applying pulsed electric fields of slightly unequal strengths. DNA fragments ranging between 61 and 209 kbp were separated within 15 s in the 'DNA prism'.

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“…This virtual routing concept was further refined by Haeberle et al, who successfully extracted DNA from calf thymus using silica beads by alternating the sense of rotation 24 . Automated extraction of human genomic DNA was demonstrated by Kloke et al 25 , who implemented novel ball-pen pierceable seals to route the sample lysate through an integrated silica membrane in a Lab Tube platform. Although these routing systems are effective, they often required an external stimulus to direct the fluids.…”

Section: Introductionmentioning

confidence: 99%

Phase-selective graphene oxide membranes for advanced microfluidic flow control

et al. 2016

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For the first time, we harness the unique phase-selectivity of chip-integrated graphene oxide (GO) membranes to significantly enhance flow control on centrifugal microfluidic platforms. In this paper, we present novel processes for the assembly of these GO membranes into polymeric microfluidic systems and demonstrate that multilayer GO membranes allow the passage of water while blocking pressurized air and organic solutions.

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The LabTube – a novel microfluidic platform for assay automation in laboratory centrifuges

Abstract: Based on a centrifugally-controlled ballpen-mechanism, the LabTube enables cost-efficient assay automation at even low sample throughput using standard centrifuges.

Cited by 30 publications

References 29 publications

Centrifugal LabTube platform for fully automated DNA purification and LAMP amplification based on an integrated, low-cost heating system

Centrifugal LabTube platform for fully automated DNA purification and LAMP amplification based on an integrated, low-cost heating system

Exploiting biased reptation for continuous flow preparative DNA fractionation in a versatile microfluidic platform

Phase-selective graphene oxide membranes for advanced microfluidic flow control

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