Gene Expression-Based Molecular Test as Diagnostic Aid for the Differential Diagnosis of Psoriasis and Eczema in Formalin-Fixed and Paraffin-Embedded Tissue, Microbiopsies, and Tape Strips.

Highly effective targeted therapies are available to treat noncommunicable chronic inflammatory skin diseases. In contrast, the exact diagnosis of noncommunicable chronic inflammatory skin diseases is complicated by its complex pathogenesis and clinical and histological overlap. Particularly, the differential diagnosis of psoriasis and eczema can be challenging in some cases, and molecular diagnostic tools need to be developed to support a gold standard diagnosis. The aim of this work was to develop a real-time PCR-based molecular classifier to distinguish psoriasis from eczema in formalin-fixed and paraffin-embedded-fixed skin samples and to evaluate the use of minimally invasive microbiopsies and tape strips for molecular diagnosis. In this study, we present a formalin-fixed and paraffin-embedded-based molecular classifier that determines the probability for psoriasis with a sensitivity/specificity of 92%/100%, respectively, and an area under the curve of 0.97, delivering comparable results to our previous published RNAprotect-based molecular classifier. The psoriasis probability, as well as levels of NOS2 expression, positively correlated with the disease hallmarks of psoriasis and negatively with eczema hallmarks. Furthermore, minimally invasive tape strips and microbiopsies were effectively used to differentiate psoriasis from eczema. In summary, the molecular classifier offers broad usage in pathology laboratories as well as outpatient settings and can support the differential diagnosis of noncommunicable chronic inflammatory skin diseases on a molecular level using formalin-fixed and paraffin-embedded tissue, microbiopsies, and tape strips.

Configurable 3D Printed Microfluidic Multiport Valves with Axial Compression.

In the last decade, the fabrication of microfluidic chips was revolutionized by 3D printing. It is not only used for rapid prototyping of molds, but also for manufacturing of complex chips and even integrated active parts like pumps and valves, which are essential for many microfluidic applications. The manufacturing of multiport injection valves is of special interest for analytical microfluidic systems, as they can reduce the injection to detection dead volume and thus enhance the resolution and decrease the detection limit. Designs reported so far use radial compression of rotor and stator. However, commercially available nonprinted valves usually feature axial compression, as this allows for adjustable compression and the possibility to integrate additional sealing elements. In this paper, we transfer the axial approach to 3D-printed valves and compare two different printing techniques, as well as six different sealing configurations. The tightness of the system is evaluated with optical examination, weighing, and flow measurements. The developed system shows similar performance to commercial or other 3D-printed valves with no measurable leakage for the static case and leakages below 0.5% in the dynamic case, can be turned automatically with a stepper motor, is easy to scale up, and is transferable to other printing methods and materials without design changes.

Vertical Scanning Interferometry for Label-Free Detection of Peptide-Antibody Interactions.

Peptide microarrays are a fast-developing field enabling the mapping of linear epitopes in the immune response to vaccinations or diseases and high throughput studying of protein-protein interactions. In this respect, a rapid label-free measurement of protein layer topographies in the array format is of great interest but is also a great challenge due to the extremely low aspect ratios of the peptide spots. We have demonstrated the potential of vertical scanning interferometry (VSI) for a detailed morphological analysis of peptide arrays and binding antibodies. The VSI technique is shown to scan an array area of 5.1 square millimeters within 3⁻4 min at a resolution of 1.4 µm lateral and 0.1 nm vertical in the full automation mode. Topographies obtained by VSI do match the one obtained by AFM measurements, demonstrating the accuracy of the technique. A detailed topology of peptide-antibody layers on single spots was measured. Two different measurement regions are distinguished according to the antibody concentration. In the case of weakly diluted serum, the thickness of the antibody layer is independent of the serum dilution and corresponds to the physical thickness of the accumulated antibody layer. In strongly diluted serum, the thickness measured via VSI is linearly proportional to the serum dilution.