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Knowledge about individual daily herbage dry matter (DM) intake (DMI) helps identifying efficient dairy cows and adapting supplementation better to herbage intake and nutrient requirements of grazing dairy cows. With the aid of behavioural characteristics, raw data recorded with the RumiWatch (RW) system and processed with the RW converter 0.7.3.31 (C31), estimation of herbage DMI may be possible. First, C31, which allows differentiation of prehension bites and mastication chews, was validated through direct observation of behavioural characteristics and compared to the previous RW converter 0.7.3.11 (C11). Further, the influence of a low and high pre-grazing herbage mass (HM), with the same target herbage allowance (HA), on bite mass, DMI, number of prehension bites, and milk production was investigated. In total, 24 lactating Holstein cows were pairwise allotted to one of two HM treatments. The cows received a new pasture paddock twice per day with a daily target HA of 22 kg DM per cow/day. On average, low HM (LHM) and high HM (HHM) paddocks had an HM of 589 and 2288 kg DM/ha, respectively, above 6.7 click units (1 CU = 0.5 cm). Overall, LHM cows produced 2.7 kg/day more milk and 2.5 kg/day more energy-corrected milk, had the same herbage DMI and a similar prehension bite mass. The averaged bite mass per week was 0.49 g DM/bite (LHM) or 0.47 g DM/bite (HHM), respectively. A longer eating time (617 vs. 559 min/day) and a shorter rumination time (297 vs. 365 min/day) were observed for the LHM cows compared with the HHM cows. The validation of the RW showed similar results for C11 and C31 apart from prehension bites, where C31 showed a mean absolute deviation of 12.4%. Pre-grazing HM had no effect on relevant behavioural characteristics for prospective intake estimation, namely, bite mass and number of prehension bites.

Accurately measuring flow rates is a key requirement in many medical applications such as infusion and drug delivery systems. A major drawback of current systems is the low resolution of the sensors in the low flow rate regime. In this article, we present a method based on Holographic PIV/PTV that has been used for the first time to measure flow rates in the range of a few nL/min. Our method requires a very simple setup that combines lensless holography with particle velocimetry. For flow rates in the 70 to 200 nL/min range, the highest uncertainty was 5.6% (coverage factor k=2). This is an open-source project; the CAD designs and software source code are available at https://github.com/gui-miotto/holovel.

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.

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 (, n = 4 LabDisks) of the time to positivity (tp) of the LAMP reactions (n = 11 replicates per LabDisk) is taken. The 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.

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 co-transport. 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.

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.

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

Automated and robust separation of 14 μl of plasma from 40 μl of whole blood at a purity of 99.81% ± 0.11% 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.

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.

Within a project sponsored by DARA, 28 ERS-1 SLC (single look complex) images of an agricultural area in the upper Rhine Valley were processed and integrated into a GIS structure. The GIS contains information on field-boundaries, land-use, soils, and results of continuous soil-moisture and rainfall measurements on several fields in the test site (approx. 50 km2). The GIS is used to establish a relationship between ERS-1 backscatter and measured surface soil-moisture for different land-uses. Since backscatter depends on soil and surface properties, statistical relations between temporal backscatter profiles of different soil- surface complexes are established. They allow us to normalize the multitemporal ERS-1 images to a reference backscatter behavior of corn on loamy sand. Since surface soil-moisture was measured for this soil-surface complex, the ERS-1 images can be converted into reference surface-moisture images. These images are interpreted. Irrigated areas can be identified by their high reference surface-moisture.© (1995) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.