Markus Grumann

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.

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%.

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...

Integrated Sample Preparation, Reaction, and Detection on a High-Frequency Centrifugal Microfluidic Platform

We extend the toolbox of lab procedures in life sciences by development of centrifugal microfluidics for high-level process integration. This is accomplished by implementing novel functional principles for sedimentation, batch-mode mixing, frequency-dependent online flow control, and optical read-out, which can be integrated into a process chain. The modular centrifugal setup comprises a microstructured disposable polymer disk as well as a reusable spinning and detection unit. We successfully developed centrifugal microfluidic technologies, which are suitable for sample preparation, process engineering, personalized diagnostics, and hematology, on this platform.

Parallelization of chip-based fluorescence immuno-assays with quantum-dot labeled beads

This paper presents an optical concept for the read-out of a parallel, bead-based fluorescence immunoassay conducted on a lab-on-a-disk platform. The reusable part of the modular setup comprises a detection unit featuring a single LED as light source, two emission-filters, and a color CCD-camera as standard components together with a spinning drive as actuation unit. The miniaturized lab-on-a-disk is devised as a disposable. In the read-out process of the parallel assay, beads are first identified by the color of incorporated quantum dots (QDs). Next, the reaction-specific fluorescence signal is quantified with FluoSpheres-labeled detection anti-bodies. To enable a fast and automated read-out, suitable algorithms have been implemented in this work. Based on this concept, we successfully demonstrated a Hepatitis-A assay on our disk-based lab-on-a-chip.

Aggregation of bead-monolayers in flat microfluidic chambers – simulation by the model of porous media

In this paper, we for the first time simulate the process of hydrodynamic bead aggregation in a flat micro-fluidic chamber by a porous-media model in an iterative routine. This allows us to optimize the chamber design of our recently developed experimental method to form periodical monolayers from the flow of bead suspension. Periodical monolayers are advantageous for parallel assay formats since they enhance the mechanical rigidity of the aggregated pattern. This is important to avoid a spatial rearrangement along various steps of a read-out procedure which would impair the correlation between measurements. Furthermore, the monolayer formation guarantees the individual optical accessibility of all probe beads. By modelling the monolayers with porous media, we can drastically reduce the degrees of freedom in a two-phase, multi-particle problem. This way, we are able to compute stationary hydrodynamic flow patterns in the chamber. In order to simulate the complete filling process from these stationary solutions, we developed an iterative master routine which takes the transient aggregation pattern as the initial condition, then evaluates the placement of the newly introduced beads, and finally converts the points of aggregation into porous media.

Design and Fabrication of a Centrifugally Driven Microfluidic Disk for Fully Integrated Metabolic Assays on Whole Blood

For the first time, we present a novel and fully integrated centrifugal microfluidic “ lab-on-a-disk” for rapid metabolic assays in human whole blood. All essential steps comprising blood sampling, metering, plasma extraction and the final optical detection are conducted within t = 150 s in passive structures integrated on one disposable disk. Our technology features a novel plasma extraction structure (V = 500 nL, CV < 5%) without using any hydrophobic microfluidics where the purified plasma (cRBC< 0.11%) is centrifugally separated and subsequently extracted through a capillarily primed extraction channel into the detection chamber. While this capillary extraction requires precisely defined, narrow micro-structures, the reactive mixing and detection is most efficient within larger cavities. The corresponding manufacturing technique of these macro- and micro structures in the range of 30 µ m to 1000 µ m is also presented for the first time: A novel, cost-efficient hybrid prototyping technique of a multiscale epoxy master for subsequent hot embossing of polymer disks.

FORMATION OF HEXAGONAL MONOLAYERS BY FLOW OF BEAD SUSPENSIONS INTO FLAT MICROCHAMBERS

To realize a highly parallel optical detection in bead-based bioanalytical assays, we investigate the hydrodynamic aggregation of bead suspensions in a hexagonally periodical monolayer by a pressure-driven flow through a microfluidic structure. This device consists of one inlet channel connected to a shallow chamber with a depth that only slightly exceeds the diameter of the beads. To enforce the aggregation of the beads, the flow leaves the chamber via outlet channels possessing a depth smaller than a bead diameter. This way the outlets act as barriers to the beads and force them to accumulate in the chamber. Benchmarking different chamber and outlet designs we found an optimum filing behavior for a rhombus-like aggregation chamber connected to a single outlet channel at the same width as the chamber. Here, the aperture angle of 60° fosters hexagonal aggregation patterns which leads to the highest packaging density. Reproducible filling ratios of more than 94% have been achieved. The rhombus-like chamber also shows the shows the smallest increase of the hydrodynamic resistance during filling and the best rinsing behavior which allows to minimize the volume of washing detergents used for a bioassay. Zones of accumulated beads redistribute the hydrodynamic flow through the device during the filling process. CFD-simulations, embedded in an iterative master-routine, are carried out to describe the complete process of filling and to assist the process of design optimization.© 2003 ASME

Optimized Creation of Monolayers for Parallel Readout of Bead-Based Assays

This publication presents the basic studies and the design optimization process of a microfluidic aggregation chamber in order to achieve highly periodical and mechanically stable monolayers of beads. All investigated designs of microfluidic devices exhibit one inlet channel connected to the so-called aggregation chamber. This chamber measures a depth which only slightly exceeds the diameter of the beads and possesses several outlet channels. With depths of the outlets smaller than a bead diameter, the beads are kept back and forced to accumulate inside the aggregation chamber as a mono-layer. CFD-simulations are carried out to describe the process of filling and to assist the process of design optimization.