Validation of a novel, fully integrated and flexible microarray benchtop facility for gene expression profiling.

Here we describe a novel microarray platform that integrates all functions needed to perform any array-based experiment in a compact instrument on the researcher's laboratory benchtop. Oligonucle otide probes are synthesized in situ via a light- activated process within the channels of a three-dimensional microfluidic reaction carrier. Arrays can be designed and produced within hours according to the user's requirements. They are processed in a fully automatic workflow. We have characterized this new platform with regard to dynamic range, discrimination power, reproducibility and accuracy of biological results. The instrument detects sample RNAs present at a frequency of 1:100 000. Detection is quantitative over more than two orders of magnitude. Experiments on four identical arrays with 6398 features each revealed a mean coefficient of variation (CV) value of 0.09 for the 6398 unprocessed raw intensities indicating high reproducibility. In a more elaborate experiment targeting 1125 yeast genes from an unbiased selection, a mean CV of 0.11 on the fold change level was found. Analyzing the transcriptional response of yeast to osmotic shock, we found that biological data acquired on our platform are in good agreement with data from Affymetrix GeneChips, quantitative real-time PCR and--albeit somewhat less clearly--to data from spotted cDNA arrays obtained from the literature.

[Thigh abscess as first manifestation of a sigma carcinoma]. / Oberschenkelabszedierung als Erstmanifestation eines metastasierten Sigmakarzinoms.

MEDICAL HISTORY: An extensive thigh abscess was the reason for a hospital admission of a 66 year old woman. Apart from that there were no other known preexisting diseases. INVESTIGATIONS: The reason for the abscess was a perforated sigma carcinoma, which was incidentally found. Treatments and course: We treated the patient with a sigma resection and several debridements of the thigh. But for all that she died from septic shock. CONCLUSION: This case illustrates the importance of cancer screening programs and an accurate investigation of the medical history and clinical examination. This can be supported by a failure management, which does not sanction a failure but considers it as a chance for future improvement.

Enrichment of viable bacteria in a micro-volume by free-flow electrophoresis.

Macro- to micro-volume concentration of viable bacteria is performed in a microfluidic chip. The enrichment principle is based on free flow electrophoresis and is demonstrated for Gram positive bacteria. Bacteria from a suspension flow are trapped on a gel interface that separates the trapping location from integrated actuation electrodes in order to enable non-destructive trapping. The microfluidic chip contains integrated electrolytic gas expulsion structures and phaseguides for gel and liquid handling. Trapping efficiency is systematically optimized to reach 25 times the initial concentration from a theoretical maximum of 30. Finally, enrichment from analytically relevant concentrations down to 3 × 10(2) colony forming units per millilitre is demonstrated with a trapping efficiency of 80% which represents the most important parameter in enrichment.

Phaseguides: a paradigm shift in microfluidic priming and emptying.

Phaseguide technology gives complete control over filling and emptying of any type of microfluidic structures, independent of the chamber and channel geometry. The technique is based on a step-wise advancement of the liquid-air interface using the meniscus pinning effect. In this paper, the main effects and parameters underlying the phaseguiding principle are discussed and a demonstration is given of its potential for dead angle filling, spatially controlled phaseguide overflow and sequential phaseguide overflow, all accumulating in a passive valving approach. Phaseguides represent a new direction in microfluidic design thinking that will prove a leap forward towards more simple, flexible and reliable microfluidic systems.

Microfluidic concentration of bacteria by on-chip electrophoresis.

In this contribution, we present a system for efficient preconcentration of pathogens without affecting their viability. Development of miniaturized molecular diagnostic kits requires concentration of the sample, molecule extraction, amplification, and detection. In consequence of low analyte concentrations in real-world samples, preconcentration is a critical step within this workflow. Bacteria and viruses exhibit a negative surface charge and thus can be electrophoretically captured from a continuous flow. The concept of phaseguides was applied to define gel membranes, which enable effective and reversible collection of the target species. E. coli of the strains XL1-blue and K12 were used to evaluate the performance of the device. By suppression of the electroosmotic flow both strains were captured with efficiencies of up to 99%. At a continuous flow of 15 µl/min concentration factors of 50.17 ± 2.23 and 47.36 ± 1.72 were achieved in less than 27 min for XL1-blue and K12, respectively. These results indicate that free flow electrophoresis enables efficient concentration of bacteria and the presented device can contribute to rapid analyses of swab-derived samples.