Jens Ducrée

Batch-mode mixing on centrifugal microfluidic platforms

We present two novel fluidic concepts to drastically accelerate the process of mixing in batch-mode (stopped-flow) on centrifugal microfluidic platforms. The core of our simple and robust setup exhibits a microstructured disk with a round mixing chamber rotating on a macroscopic drive unit. In the first approach, magnetic beads which are prefilled into the mixing chamber are periodically deflected by a set of permanent magnets equidistantly aligned at spatially fixed positions in the lab-frame. Their radial positions alternatingly deviate by a slight positive and negative offset from the mean orbit of the chamber to periodically deflect the beads inbound and outbound during rotation. Advection is induced by the relative motion of the beads with respect to the liquid which results from the magnetic and centrifugal forces, as well as inertia. In a second approach--without magnetic beads--the disk is spun upon periodic changes in the sense of rotation. This way, inertia effects induce stirring of the liquids. As a result, both strategies accelerate mixing from about 7 minutes for mere diffusion to less than five seconds. Combining both effects, an ultimate mixing time of less than one second could be achieved.

Centrifugal extraction of plasma from whole blood on a rotating disk

We present a centrifugal process for the extraction of plasma from sediment by a decanting structure, terminating with metered plasma which is readily available for subsequent on-disk processing. Our technique supplies 2 microl plasma from 5 microl of whole blood at moderate spinning frequencies of 40 Hz within 20 s, only. The residual cell concentration in the purified plasma amounts to less than 0.11%, independent of the frequency of rotation. A capillary duct connects the extracted plasma to subsequent on-disk processing units.

Rapid prototyping of microfluidic chips in COC

We present a novel, cost-efficient process chain for fast tooling and small-lot replication of high-quality, multi-scale microfluidic polymer chips within less than 5 days. The fabrication chain starts with a primary master which is made by well-established cleanroom processes such as DRIE or negative SU-8 resist based surface micromachining. The formation of undercuts in the master which would complicate demolding is carefully avoided. Secondary PDMS masters or epoxy-based masters which are more suitable for common polymer replication schemes such as soft-embossing, hot-embossing or injection molding are subsequently cast from the primary masters. The polymer replica are mainly made of COC and show excellent fidelity with the conventionally micromachined master while displaying no degeneration, even after more than 200 cycles. The use of other polymers such as PMMA is also possible. The process chain further includes surface modification techniques for overall, long-term stable hydrophilic coatings and for local hydrophobic patches as well as a durable sealing based on thermal bonding.

Fully integrated whole blood testing by real-time absorption measurement on a centrifugal platform

We present a novel microfluidic concept to enable a fast colorimetric alcohol assay from a single droplet of whole blood. The reduced turn-around time of 150 seconds is, on the one hand, achieved by a full process integration including metering, mixing with reagents, and sedimentation of cellular constituents. On the other hand, our novel total internal reflection (TIR) scheme allows to monitor the increase of the absorbance values in real-time. Thus, the saturation values can be predicted accurately based on an extrapolation of real-time measurements acquired during a 100 second initial period of rotation. Additionally, we present a metering structure to define nanolitre sample volumes at a coefficient of variation (CV) below 5%.

Frequency-dependent transversal flow control in centrifugal microfluidics.

This work presents a novel flow switch for centrifugal microfluidic platforms which is solely controlled by the Coriolis pseudo force. This Coriolis switch consists of an inverse Y-structure with one common upstream channel and two symmetric outlets on a rotating disk. Above a certain threshold frequency, the Coriolis force becomes dominant that the entire flow is diverted into one of the outlets which is selected by the direction of rotation. The threshold frequency has been measured to be 350 rad s(-1)(approximately 55.7 Hz) for a channel width of 360 microm and a depth of 125 microm. The results are supported by extensive CFD simulations.

Visualization of flow patterning in high-speed centrifugal microfluidics

This work presents a new experimental setup for image capturing of centrifugally driven flows in disk-based microchannels rotating at high frequencies of up to 150Hz. To still achieve a micron-scale resolution, smearing effects are minimized by a microscope-mounted CCD camera featuring an extremely short minimum exposure time of 100ns, only. The image capture is controlled by a real-time PC board which sends delayed trigger signals to the CCD camera and to a stroboscopic flash upon receiving the zero-crossing signal of the rotating disk. The common delay of the trigger signals is electronically adjusted according to the spinning frequency. This appreciably improves the stability of the captured image sequences. Another computer is equipped with a fast framegrabber PC board to directly acquire the image data from the CCD camera. A maximum spatial resolution ranging between 4.5μm at rest and 10μm at a 150Hz frequency of rotation is achieved. Even at high frequencies of rotation, image smearing does not sign...

Integration of functional materials and surface modification for polymeric microfluidic systems

The opportunity for the commercialization of microfluidic systems has surged over the recent decade, primarily for medical and the life science applications. This positive development has been spurred by an increasing number of integrated, highly functional lab-on-a-chip technologies from the research community. Toward commercialization, there is a dire need for economic manufacture which involves optimized cost for materials and structuring on the front-end as well as for a range of back-end processing steps such as surface modification, integration of functional elements, assembly and packaging. Front-end processing can readily resort to very well established polymer mass fabrication schemes, e.g. injection molding. Also assembly and packaging can often be adopted from commercially available processes. In this review, we survey the back-end processes of hybrid material integration and surface modification which often need to be tailored to the specifics of miniaturized polymeric microfluidic systems. On the one hand, the accurate control of these back-end processes proves to be the key to the technical function of the system and thus the value creation. On the other hand, the integration of functional materials constitutes a major cost factor. (Some figures may appear in colour only in the online journal)

Platelet Adhesion and Degranulation Induce Pro-Survival and Pro-Angiogenic Signalling in Ovarian Cancer Cells

Thrombosis is common in ovarian cancer. However, the interaction of platelets with ovarian cancer cells has not been critically examined. To address this, we investigated platelet interactions in a range of ovarian cancer cell lines with different metastatic potentials [HIO-80, 59M, SK-OV-3, A2780, A2780cis]. Platelets adhered to ovarian cancer cells with the most significant adhesion to the 59M cell line. Ovarian cancer cells induced platelet activation [P-selectin expression] in a dose dependent manner, with the most significant activation seen in response to the 59M cell line. The platelet antagonists [cangrelor, MRS2179, and apyrase] inhibited 59M cell induced activation suggesting a P2Y12 and P2Y1 receptor mediated mechanism of platelet activation dependent on the release of ADP by 59M cells. A2780 and 59M cells potentiated PAR-1, PAR-4, and TxA2 receptor mediated platelet activation, but had no effect on ADP, epinephrine, or collagen induced activation. Analysis of gene expression changes in ovarian cancer cells following treatment with washed platelets or platelet releasate showed a subtle but valid upregulation of anti-apoptotic, anti-autophagy pro-angiogenic, pro-cell cycle and metabolic genes. Thus, ovarian cancer cells with different metastatic potential adhere and activate platelets differentially while both platelets and platelet releasate mediate pro-survival and pro-angiogenic signals in ovarian cancer cells.

Centrifugal microfluidics for cell analysis.

Over the past two decades, centrifugal microfluidic systems have successfully demonstrated their capability for robust, high-performance liquid handling to enable modular, multi-purpose lab-on-a-chip platforms for a wide range of life-science applications. Beyond the handling of homogeneous liquids, the unique, rotationally controlled centrifugal actuation has proven to be specifically advantageous for performing cell and particle handling and assays. In this review we discuss technologies to implement two important steps for cell handling, namely separation and capturing/counting.

Array-based capture, distribution, counting and multiplexed assaying of beads on a centrifugal microfluidic platform

We present a novel centrifugal microfluidic platform for the highly efficient manipulation and analysis of particles for applications in bead-based assays. The platform uses an array of geometrical V-cup barriers to trap particles using stopped-flow sedimentation under highly reproducible hydrodynamic conditions. The impact parameters governing the occupancy distribution and capture efficiency of the arrayed traps are investigated. The unique, nearly 100% capture efficiency paired with the capability to establish sharply peaked, single occupancy distributions enables a novel, digital readout mode for color-multiplexed, particle-based assays with low-complexity instrumentation. The presented technology marks an essential step towards a versatile platform for the integration of bead- and cell-based biological assays.