Knowledge about Hand Hygiene and Related Infectious Disease Awareness among Primary School Children in Germany.

Hand hygiene is a cornerstone of infection prevention. However, few data are available for school children on their knowledge of infectious diseases and their prevention. The aim of the study was to develop and apply a standardized questionnaire for children when visiting primary schools to survey their knowledge about infectious diseases, pathogen transmission and prevention measures. Enrolling thirteen German primary schools, 493 questionnaires for grade three primary school children were included for further analyses, comprising 257 (52.1%) girls and 236 (47.9%) boys with an age range of 8-11 years. Out of 489 children, 91.2% participants indicated that they knew about human-to-human transmissible diseases. Of these, 445 children responded in detail, most frequently mentioning respiratory and gastrointestinal diseases, followed by childhood diseases. Addressing putative hygiene awareness-influencing factors, it was worrisome that more than 40.0% of the children avoided visiting the sanitary facilities at school. Most of the children (82.9%) noted that they did not like to use the sanitary facilities at school because of their uncleanliness and the poor hygienic behavior of their classmates. In conclusion, basic infection awareness exists already in primary school age children. Ideas about the origin and prevention of infections are retrievable, however, this knowledge is not always accurate and adequately contextualized. Since the condition of sanitary facilities has a strong influence on usage behavior, the child's perspective should be given more consideration in the design and maintenance of sanitary facilities.

A Label-Free Optical Detection of Pathogens in Isopropanol as a First Step towards Real-Time Infection Prevention.

The detection of pathogens is a major public health issue. Every year, thousands of people die because of nosocomial infections. It is therefore important to be able to detect possible outbreaks as early as possible, especially in the hospital environment. Various pathogen detection techniques have already been demonstrated. However, most of them require expensive and specific equipment, and/or complex protocols, which, most of the time, involve biochemical reaction and labelling steps. In this paper, a new method that combines microscopic imaging and machine learning is described. The main benefits of this approach are to be low-cost, label-free and easy to integrate in any suitable medical device, such as hand hygiene dispensers. The suitability of this pathogen detection method is validated using four bacteria, both in PBS (Phosphate Buffered Saline) and in isopropanol. In particular, we demonstrated an efficient pathogenic detection that is sensible to changes in the composition of a mixture of pathogens, even in alcohol-based solutions.

Elasticity mapping of pore-suspending native cell membranes.

The mechanics of cellular membranes are governed by a non-equilibrium composite framework consisting of the semiflexible filamentous cytoskeleton and extracellular matrix proteins linked to the lipid bilayer. While elasticity information of plasma membranes has mainly been obtained from whole cell analysis, techniques that allow addressing local mechanical properties of cell membranes are desirable to learn how their lipid and protein composition is reflected in the elastic behavior on local length scales. Introduced here is an approach based on basolateral membranes of polar epithelial Madin-Darby canine kidney (MDCK) II cells, prepared on a highly ordered porous substrate that allows elastic mapping on a submicrometer-length scale. A strong correlation between the density of actin filaments and the measured membrane elasticity is found. Spatially resolved indentation experiments carried out with atomic force and fluorescence microscope permit relation of the supramolecular structure to the elasticity of cellular membranes. It is shown that the elastic response of the pore spanning cell membranes is governed by local bending modules rather than lateral tension.

Continuous high throughput nanofluidic separation through tangential-flow vertical nanoslit arrays.

Nanofluidic devices exhibit unique, tunable transport properties that may lead to breakthroughs in molecular separations and sensing. However, the throughput of these devices is orders of magnitude too small for the processing of macroscopic samples. Here we overcome this problem by combining two technological innovations. First, nanofluidic channels are made as vertical slits connecting the two sides of a silicon nitride membrane. Arbitrary arrays of such nanoslits down to 15 nm wide with <6 Å uniformity were made by merging the idea of templating with chemical mechanical polishing to create a scalable, nanolithography-free wafer level process. Second, we provide for efficient solute transport to and from the openings of the nanoslits by incorporating the nanofluidic membrane into a microfluidic tangential-flow system, which is also fabricated at wafer level. As an exemplary application, we demonstrate charge-based continuous flow separation of small molecules with a selectivity of 100 and constant flux over more than 100 hours of operation. This proves the exciting possibility of exploiting transport phenomena governed by precision-engineered nanofluidic devices at a macroscopic scale.

Cytoskeleton remodelling of confluent epithelial cells cultured on porous substrates.

The impact of substrate topography on the morphological and mechanical properties of confluent MDCK-II cells cultured on porous substrates was scrutinized by means of various imaging techniques as well as atomic force microscopy comprising force volume and microrheology measurements. Regardless of the pore size, ranging from 450 to 5500 nm in diameter, cells were able to span the pores. They did not crawl into the holes or grow around the pores. Generally, we found that cells cultured on non-porous surfaces are stiffer, i.e. cortical tension rises from 0.1 to 0.3 mN m(-1), and less fluid than cells grown over pores. The mechanical data are corroborated by electron microscopy imaging showing more cytoskeletal filaments on flat samples in comparison to porous ones. By contrast, cellular compliance increases with pore size and cells display a more fluid-like behaviour on larger pores. Interestingly, cells on pores larger than 3500 nm produce thick actin bundles that bridge the pores and thereby strengthen the contact zone of the cells.

Nanostructured cavity devices for extracellular stimulation of HL-1 cells.

Microelectrode arrays (MEAs) are state-of-the-art devices for extracellular recording and stimulation on biological tissue. Furthermore, they are a relevant tool for the development of biomedical applications like retina, cochlear and motor prostheses, cardiac pacemakers and drug screening. Hence, research on functional cell-sensor interfaces, as well as the development of new surface structures and modifications for improved electrode characteristics, is a vivid and well established field. However, combining single-cell resolution with sufficient signal coupling remains challenging due to poor cell-electrode sealing. Furthermore, electrodes with diameters below 20 µm often suffer from a high electrical impedance affecting the noise during voltage recordings. In this study, we report on a nanocavity sensor array for voltage-controlled stimulation and extracellular action potential recordings on cellular networks. Nanocavity devices combine the advantages of low-impedance electrodes with small cell-chip interfaces, preserving a high spatial resolution for recording and stimulation. A reservoir between opening aperture and electrode is provided, allowing the cell to access the structure for a tight cell-sensor sealing. We present the well-controlled fabrication process and the effect of cavity formation and electrode patterning on the sensor's impedance. Further, we demonstrate reliable voltage-controlled stimulation using nanostructured cavity devices by capturing the pacemaker of an HL-1 cell network.

A µ-biomimetic flow sensor for medical and pharmaceutical applications.

Flow sensing is pivotal in many medical and pharmaceutical applications. Most commercial flow sensors are either expensive, complex, or consume a lot of energy, while low cost sensors usually lack sensitivity, robustness, or long-term stability. In addition, the maintenance and sterilization of most commercial flow sensors is difficult to perform. Here, we present a new µ-biomimetic flow sensor based on the fish lateral line. It measures flow velocity and detects the transition between laminar and turbulent flow, thereby fulfilling most requirements for medical and pharmaceutical applications. Additionally, it has a modular setup featuring a screened or passive bypass configuration, enabling it not only to meter flow in medical applications but also under harsh or well-defined environmental conditions, such as found in pharmaceutical applications. The sensor is robust and can be easily cleaned. Individual parts of the sensor can even be replaced or sterilized. In sum, this sensor opens up a whole new field of applications in the area of medical and pharmaceutical related flow monitoring.

The development of a µ-biomimetic uncooled IR-Sensor inspired by the infrared receptors of Melanophila acuminata.

The beetle Melanophila acuminata uses a specialized organ to detect infrared radiation. The organ consists of about 100 individual sensilla. The main component of the sensillum is a pressure chamber. Upon absorption of radiation, the pressure increases, and the tip of a dendrite is deformed. A unique feature of the organ is a compensation mechanism that prevents large pressures. The beetle uses this organ to detect forest fires and to navigate inside burning woods. However, the sensitivity is part of a long-lasting discussion, providing thresholds between [Formula: see text] and [Formula: see text]. To end the decade-long discussion and to provide a novel type of infrared sensor, we are developing an uncooled µ-biomimetic infrared (IR) sensor inspired by Melanophila acuminata using MEMS technology. Here, we present the development of a µ-capacitor that is used to detect pressure changes and the characterization of the compensation mechanism. We describe the microtechnological fabrication process for air-filled capacitors with a ratio of diameter-to-electrode distance of 1000 and a technique to fill the sensor bubble-free with water. Finally, we estimate the sensitivity of the beetle using a theoretical model of the sensillum.

µ-Biomimetic flow-sensors--introducing light-guiding PDMS structures into MEMS.

In the area of biomimetics, engineers use inspiration from natural systems to develop technical devices, such as sensors. One example is the lateral line system of fish. It is a mechanoreceptive system consisting of up to several thousand individual sensors called neuromasts, which enable fish to sense prey, predators, or conspecifics. So far, the small size and high sensitivity of the lateral line is unmatched by man-made sensor devices. Here, we describe an artificial lateral line system based on an optical detection principle. We developed artificial canal neuromasts using MEMS technology including thick film techniques. In this work, we describe the MEMS fabrication and characterize a sensor prototype. Our sensor consists of a silicon chip, a housing, and an electronic circuit. We demonstrate the functionality of our µ-biomimetic flow sensor by analyzing its response to constant water flow and flow fluctuations. Furthermore, we discuss the sensor robustness and sensitivity of our sensor and its suitability for industrial and medical applications. In sum, our sensor can be used for many tasks, e.g. for monitoring fluid flow in medical applications, for detecting leakages in tap water systems or for air and gas flow measurements. Finally, our flow sensor can even be used to improve current knowledge about the functional significance of the fish lateral line.

Towards an optimum design for electrostatic phase plates.

Charging of physical phase plates is a problem that has prevented their routine use in transmission electron microscopy of weak-phase objects. In theory, electrostatic phase plates are superior to thin-film phase plates since they do not attenuate the scattered electron beam and allow freely adjustable phase shifts. Electrostatic phase plates consist of multiple layers of conductive and insulating materials, and are thus more prone to charging than thin-film phase plates, which typically consist of only one single layer of amorphous material. We have addressed the origins of charging of Boersch phase plates and show how it can be reduced. In particular, we have performed simulations and experiments to analyze the influence of the insulating Si3N4 layers and surface charges on electrostatic charging. To optimize the performance of electrostatic phase plates, it would be desirable to fabricate electrostatic phase plates, which (i) impart a homogeneous phase shift to the unscattered electrons, (ii) have a low cut-on frequency, (iii) expose as little material to the intense unscattered beam as possible, and (iv) can be additionally polished by a focused ion-beam instrument to eliminate carbon contamination accumulated during exposure to the unscattered electron beam (Walter et al., 2012, Ultramicroscopy, 116, 62-72). We propose a new type of electrostatic phase plate that meets the above requirements and would be superior to a Boersch phase plate. It consists of three free-standing coaxial rods converging in the center of an aperture (3-fold coaxial phase plate). Simulations and preliminary experiments with modified Boersch phase plates indicate that the fabrication of a 3-fold coaxial phase plate is feasible.

Microfluidic cryofixation for correlative microscopy.

Cryofixation yields outstanding ultrastructural preservation of cells for electron microscopy, but current methods disrupt live cell imaging. Here we demonstrate a microfluidic approach that enables cryofixation to be performed directly in the light microscope with millisecond time resolution and at atmospheric pressure. This will provide a link between imaging/stimulation of live cells and post-fixation optical, electron, or X-ray microscopy.

A model for µ-biomimetic thermal infrared sensors based on the infrared receptors of Melanophila acuminata.

Beetles of the genus Melanophila acuminata detect forest fires from distances as far as 130 km with infrared-sensing organs. Inspired by this extremely sensitive biological device, we are developing an IR sensor that operates at ambient temperature using MEMS technology. The sensor consists of two liquid-filled chambers that are connected by a micro-fluidic system. Absorption of IR radiation by one of these chambers leads to heating and expansion of a liquid. The increasing pressure deflects a membrane covered by one electrode of a plate capacitor. The micro-fluidic system and the second chamber represent a fluidic low-pass filter, preventing slow, but large pressure changes. However, the strong frequency dependence of the filter demands a precise characterization of its properties. Here, we present a theoretical model that describes the frequency-dependent response of the sensor based on material properties and geometrical dimensions. Our model is divided into four distinct parts that address different aspects of the sensor. The model describes the frequency-dependent behaviour of the fluidic filter and a thermal low-pass filter as well as saturation effects at low frequencies. This model allows the calculation of optimal design parameters, and thereby provides the foundation for the development of such a sensor.

A fluorescence based method for the quantification of surface functional groups in closed micro- and nanofluidic channels.

Surface analysis is critical for the validation of microfluidic surface modifications for biology, chemistry, and physics applications. However, until now quantitative analytical methods have mostly been focused on open surfaces. Here, we present a new fluorescence imaging method to directly measure the surface coverage of functional groups inside assembled microchannels over a wide dynamic range. A key advance of our work is the elimination of self-quenching to obtain a linear signal even with a high density of functional groups. This method is applied to image the density and monitor the stability of vapor deposited silane layers in bonded silicon/glass micro- and nanochannels.

Advantages of voice reproduction and the development of a biomimetic self-regulating double-clack valve for a prosthesis of the larynx--a feasibility study.

The human larynx is a versatile organ. Main functions are phonation, protection and regulation of the air ways. Patients suffer severely from the diagnosis of a laryngeal carcinoma of the stages T3 and T4. In most cases this diagnosis will lead to a total laryngectomy, which is usually dissatisfying in the sense of postoperative rehabilitation. The postoperative consequences include the loss of the native voice, the loss of regular air ways via mouth and nose, sense of smell, and the inability to build up an abdominal pressure. In this paper we focus on the feasibility of a modular larynx prosthesis which enables the laryngectomee to talk with his native voice, to breathe via the regular air ways, and to build up abdominal pressure. In particular we will give insights for a postoperative solution - a modular prosthesis based on a biomimetic self-regulating double clack-valve and on a voice reconstruction module, a so called vocoder. The vocoder is a device to reproduce the natural human voice. Most important for the use is an additional device required to analyze, conserve and manage voice characteristics of the patient before surgery. The self-regulating double clack-valve is designed to build up an abdominal pressure e.g. to cough. Therefore, our valve-system is working in both directions - a two-way valve system. By bridging the gap of the regular air ways lost by laryngectomy, the sense of smell and taste are restored. In the following we will present details and characteristics of these two main components required for a modular prosthesis of the larynx in laryngectomees.

Practical aspects of Boersch phase contrast electron microscopy of biological specimens.

Implementation of physical phase plates into transmission electron microscopes to achieve in-focus contrast for ice-embedded biological specimens poses several technological challenges. During the last decade several phase plates designs have been introduced and tested for electron cryo-microscopy (cryoEM), including thin film (Zernike) phase plates and electrostatic devices. Boersch phase plates (BPPs) are electrostatic einzel lenses shifting the phase of the unscattered beam by an arbitrary angle. Adjusting the phase shift to 90° achieves the maximum contrast transfer for phase objects such as biomolecules. Recently, we reported the implementation of a BPP into a dedicated phase contrast aberration-corrected electron microscope (PACEM) and demonstrated its use to generate in-focus contrast of frozen-hydrated specimens. However, a number of obstacles need to be overcome before BPPs can be used routinely, mostly related to the phase plate devices themselves. CryoEM with a physical phase plate is affected by electrostatic charging, obliteration of low spatial frequencies, and mechanical drift. Furthermore, BPPs introduce single sideband contrast (SSB), due to the obstruction of Friedel mates in the diffraction pattern. In this study we address the technical obstacles in detail and show how they may be overcome. We use X-ray photoelectron spectroscopy (XPS) and Auger electron spectroscopy (AES) to identify contaminants responsible for electrostatic charging, which occurs with most phase plates. We demonstrate that obstruction of low-resolution features is significantly reduced by lowering the acceleration voltage of the microscope. Finally, we present computational approaches to correct BPP images for SSB contrast and to compensate for mechanical drift of the BPP.

Electrode structures for acquisition and neural stimulation controlling the cardiovascular system.

In this study we present an innovative electrode system, for many different applications in the field of cardiovascular diseases. It is a combination of intelligent communicating dry-surface electrodes, which are able to interact with different sensors especially with an invasive, ultra flexible electrode-system. Dry and smart surface electrodes, which can be integrated in textiles and therefore such electrode are almost "invisible" for patients, are used for ECG acquisition and can be integrated in a communication network. In combination with a pulse oximeter or impedance spectroscopy the pulse transit time (PTT) can be calculated. Additionally, with invasive electrodes the nervous vagus can be stimulated and therefore cardiovascular functions can be controlled. The association of an implanted stimulator with an interacting and smart monitoring system results into a cardiovascular controlling. In this work we will focus on the feasibility, suitability, fabrication and characterization of invasive and dry-surface electrode systems as a basic element and foundation for cardiovascular regulation in a closed loop.

Mechanical properties of pore-spanning lipid bilayers probed by atomic force microscopy.

We measure the elastic response of a free-standing lipid membrane to a local indentation by using an atomic force microscope. Starting point is a planar gold-coated alumina substrate with a chemisorbed 3-mercaptopropionic acid monolayer displaying circular pores of very well defined and tunable size, over which bilayers composed of N,N,-dimethyl-N,N,-dioctadecylammonium bromide or 1,2-dioleoyl-3-trimethylammonium-propane chloride were spread. Centrally indenting these "nanodrums" with an atomic force microscope tip yields force-indentation curves, which we quantitatively analyze by solving the corresponding shape equations of continuum curvature elasticity. Since the measured response depends in a known way on the system geometry (pore size, tip radius) and on material parameters (bending modulus, lateral tension), this opens the possibility to monitor local elastic properties of lipid membranes in a well-controlled setting.