The integration of a micropipette in a closed microfluidic chip with optical tweezers for investigations of single cells: erratum

In July 2011 a new concept of a closed microfluidic system equipped with a fixed micropipette, optical tweezers and a UV-Vis spectrometer was presented [Biomed. Opt. Express 2, 2299 (2011)]. Figure 1 showed falsely oriented mirrors. To clarify the design of the setup, this erratum presents a correct schematic

A combined tactile and Raman probe for tissue characterization - Design considerations

Histopathology is the golden standard for cancer diagnosis and involves the characterization of tissue components. It is labour intensive and time consuming. We have earlier proposed a combined fibre-optic near-infrared Raman spectroscopy (NIR-RS) and tactile resonance method (TRM) probe for detecting positive surgical margins as a complement to interoperative histopathology. The aims of this study were to investigate the effects of attaching an RS probe inside a cylindrical TRM sensor and to investigate how laser-induced heating of the fibre-optic NIR-RS affected the temperature of the RS probe tip and an encasing TRM sensor. In addition, the possibility to perform fibre-optic NIR-RS in a well-lit environment was investigated. A small amount of rubber latex was preferable for attaching the thin RS probe inside the TRM sensor. The temperature rise of the TRM sensor due to a fibre-optic NIR-RS at 270 mW during 20 s was less than 2 degrees C. Fibre-optic NIR-RS was feasible in a dimmed bright environment using a small light shield and automatic subtraction of a pre-recorded contaminant spectrum. The results are promising for a combined probe for tissue characterization

A Seamless Convergence of the Digital and Physical Factory Aiming in Personalized Product Emergence Process (PPEP) for Smart Products within ESB Logistics Learning Factory at Reutlingen University

AbstractA seamless convergence of the digital and physical factory aiming in personalized Product Emergence Process (PPEP) for smart products within ESB Logistics Learning Factory at Reutlingen University.A completely new business model with reference to Industrie4.0 and facilitated by 3D Experience Software in today's networked society in which customers expect immediate responses, delightful experience and simple solutions is one of the mission scenarios in the ESB Logistics Learning Factory at ESB Business School (Reutlingen University).The business experience platform provides software solutions for every organization in the company respectively in the factory. An interface with dashboards, project management apps, 3D - design and construction apps with high end visualization, manufacturing and simulation apps as well as intelligence and social network apps in a collaborative interactive environment help the user to learn the creation of a value end to end process for a personalized virtual and later real produced product.Instead of traditional ways of working and a conventional operating factory real workers and robots work semi-intuitive together. Centerpiece in the self-planned interim factory is the smart personalized product, uniquely identifiable and locatable at all times during the production process – a scooter with an individual colored mobile phone – holder for any smart phone produced with a 3D printer in lot size one. Smart products have in the future solutions incorporated internet based services – designed and manufactured - at the costs of mass products. Additionally the scooter is equipped with a retrievable declarative product memory. Monitoring and control is handled by sensor tags and a raspberry positioned on the product. The engineering design and implementation of a changeable production system is guided by a self-execution system that independently find amongst others esplanade workplaces.The imparted competences to students and professionals are project management method SCRUM, customization of workflows by Industrie4.0 principles, the enhancements of products with new personalized intelligent parts, electrical and electronic self-programmed components and the control of access of the product memory information, to plan in a digital engineering environment and set up of the physical factory to produce customer orders. The gained action-orientated experience refers to the chances and requirements for holistic digital and physical systems

Combining scanning haptic microscopy and fibre optic Raman spectroscopy for tissue characterization

The tactile resonance method (TRM) and Raman spectroscopy (RS) are promising for tissue characterization in vivo. Our goal is to combine these techniques into one instrument, to use TRM for swift scanning, and RS for increasing the diagnostic power. The aim of this study was to determine the classification accuracy, using support vector machines, for measurements on porcine tissue and also produce preliminary data on human prostate tissue. This was done by developing a new experimental set-up combining micro-scale TRM—scanning haptic microscopy (SHM)—for assessing stiffness on a micro-scale, with fibre optic RS measurements for assessing biochemical content. We compared the accuracy using SHM alone versus SHM combined with RS, for different degrees of tissue homogeneity. The cross-validation classification accuracy for healthy porcine tissue types using SHM alone was 65–81%, and when RS was added it increased to 81–87%. The accuracy for healthy and cancerous human tissue was 67–70% when only SHM was used, and increased to 72–77% for the combined measurements. This shows that the potential for swift and accurate classification of healthy and cancerous prostate tissue is high. This is promising for developing a tool for probing the surgical margins during prostate cancer surgery

CO2 gasification reactivity of char from high-ash biomass

Biomass char produced from pyrolysis processes is of great interest to be utilized as renewable solid fuels or materials. Forest byproducts and agricultural wastes are low-cost and sustainable biomass feedstocks. These biomasses generally contain high amounts of ash-forming elements, generally leading to high char reactivity. This study elaborates in detail how chemical and physical properties affect CO2 gasification rates of high-ash biomass char, and it also targets the interactions between these properties. Char produced from pine bark, forest residue, and corncobs (particle size 4–30 mm) were included, and all contained different relative compositions of ash-forming elements. Acid leaching was applied to further investigate the influence of inorganic elements in these biomasses. The char properties relevant to the gasification rate were analyzed, that is, elemental composition, specific surface area, and carbon structure. Gasification rates were measured at an isothermal condition of 800 °C with 20% (vol.) of CO2 in N2. The results showed that the inorganic content, particularly K, had a stronger effect on gasification reactivity than specific surface area and aromatic cluster size of the char. At the gasification condition utilized in this study, K could volatilize and mobilize through the char surface, resulting in high gasification reactivity. Meanwhile, the mobilization of Ca did not occur at the low temperature applied, thus resulting in its low catalytic effect. This implies that the dispersion of these inorganic elements through char particles is an important reason behind their catalytic activity. Upon leaching by diluted acetic acid, the K content of these biomasses substantially decreased, while most of the Ca remained in the biomasses. With a low K content in leached biomass char, char reactivity was determined by the active carbon surface area.publishedVersio

Large scale multifactorial likelihood quantitative analysis of BRCA1 and BRCA2 variants: An ENIGMA resource to support clinical variant classification

The multifactorial likelihood analysis method has demonstrated utility for quantitative assessment of variant pathogenicity for multiple cancer syndrome genes. Independent data types currently incorporated in the model for assessing BRCA1 and BRCA2 variants include clinically calibrated prior probability of pathogenicity based on variant location and bioinformatic prediction of variant effect, co-segregation, family cancer history profile, co-occurrence with a pathogenic variant in the same gene, breast tumor pathology, and case-control information. Research and clinical data for multifactorial likelihood analysis were collated for 1,395 BRCA1/2 predominantly intronic and missense variants, enabling classification based on posterior probability of pathogenicity for 734 variants: 447 variants were classified as (likely) benign, and 94 as (likely) pathogenic; and 248 classifications were new or considerably altered relative to ClinVar submissions. Classifications were compared with information not yet included in the likelihood model, and evidence strengths aligned to those recommended for ACMG/AMP classification codes. Altered mRNA splicing or function relative to known nonpathogenic variant controls were moderately to strongly predictive of variant pathogenicity. Variant absence in population datasets provided supporting evidence for variant pathogenicity. These findings have direct relevance for BRCA1 and BRCA2 variant evaluation, and justify the need for gene-specific calibration of evidence types used for variant classification

Large scale multifactorial likelihood quantitative analysis of BRCA1 and BRCA2 variants: An ENIGMA resource to support clinical variant classification

Abstract The multifactorial likelihood analysis method has demonstrated utility for quantitative assessment of variant pathogenicity for multiple cancer syndrome genes. Independent data types currently incorporated in the model for assessing BRCA1 and BRCA2 variants include clinically calibrated prior probability of pathogenicity based on variant location and bioinformatic prediction of variant effect, co-segregation, family cancer history profile, co-occurrence with a pathogenic variant in the same gene, breast tumor pathology, and case-control information. Research and clinical data for multifactorial likelihood analysis were collated for 1395 BRCA1/2 predominantly intronic and missense variants, enabling classification based on posterior probability of pathogenicity for 734 variants: 447 variants were classified as (likely) benign, and 94 as (likely) pathogenic; 248 classifications were new or considerably altered relative to ClinVar submissions. Classifications were compared to information not yet included in the likelihood model, and evidence strengths aligned to those recommended for ACMG/AMP classification codes. Altered mRNA splicing or function relative to known non-pathogenic variant controls were moderately to strongly predictive of variant pathogenicity. Variant absence in population datasets provided supporting evidence for variant pathogenicity. These findings have direct relevance for BRCA1 and BRCA2 variant evaluation, and justify the need for gene-specific calibration of evidence types used for variant classification. This article is protected by copyright. All rights reserved.Peer reviewe

Optical manipulation and spectroscopy of single functional globin-containing cells in microfluidic systems

Recent advances in microbiology and biomedicine require development of tools to monitor the response of single cells to environmental stimuli under controlled physiological conditions. In this thesis, I describe how this can be achieved by combining various spectroscopic techniques with optical tweezers and microfluidic systems. Apart from absorption spectroscopy the technique of resonance Raman microscopy has been used to examine the oxygenation cycle of optically trapped cells containing heme-proteins, i.e. hemoglobin in red blood cells, neuroglobin overexpressed in Escherichia coli bacteria, and the nerve globin of the polychaete annelid Aphrodite aculeate in the ganglia of the nerve chain. Resonance Raman spectroscopy is an excellent tool to investigate the structure-function relationship of heme proteins. The porphyrin group in the heme absorbs visible light and a resonance effect leads to a selective enhancement of the porphyrin vibrations, hence, the method gives information about the oxygenation as well as the spin state of the heme iron. An important issue in the field of biochemistry, biomedicine and drug discovery is the mimicking of in vivo conditions in an in vitro environment. There is a demand to monitor chemical reactions in real time with good control over the diffusion of substances. Microfluidic technology can fulfil many of these requirements. Within this work, I developed a microfluidic system equipped with a pump that is connected to a switch enabling the flushing of buffers purged with O2 or N2 to create aerobic or anaerobic conditions in a flow cell. By doing so, I was able to monitor the oxygenation cycle of heme proteins reversibly over an extended period of time, confirming that the set-up meets the demand of physiological conditions