Assessing Community Health: An Innovative Tool for Measuring Height and Length.

Anthropometric measurements, including height and length, are routinely needed for health research worldwide. Measurement boards are the current gold standard for obtaining the height and length of children. In community-based research, however, the size and weight of the measurement boards make them difficult and cumbersome to carry in the field. In addition, children and infants may express an unwillingness to be placed onto the measurement board. Electronic measuring tools commonly used in industry and contracting work are precise and portable. This study piloted a protocol to use an adapted laser measurement tool, the anthropometric measurement assist (AMA), to obtain height and recumbent length in children in Western Kenya. Intra- and inter-observer variability were determined and compared with measurement board measurements. Results of this initial pilot indicated that the AMA may be a viable alternative to measurement boards. The AMA can measure height/length accurately and reliably, is portable and is equivalent in price to measuring boards, making it a viable option for fieldwork in low-resourced countries.

A laser ablation ICP-MS based method for multiplexed immunoblot analysis: applications to manganese-dependent protein dynamics of photosystem II in barley (Hordeum vulgare L.).

Manganese (Mn) constitutes an essential co-factor in the oxygen-evolving complex of photosystem II (PSII). Consequently, Mn deficiency reduces photosynthetic efficiency and leads to changes in PSII composition. In order to study these changes, multiplexed protein assays are advantageous. Here, we developed a multiplexed antibody-based assay and analysed selected PSII subunits in barley (Hordeum vulgare L.). A selection of antibodies were labelled with specific lanthanides and immunoreacted with thylakoids exposed to Mn deficiency after western blotting. Subsequently, western blot membranes were analysed by laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS), which allowed selective and relative quantitative analysis via the different lanthanides. The method was evaluated against established liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) methods, based on data-dependent acquisition (DDA) and selected reaction monitoring (SRM). Manganese deficiency resulted in a general decrease in PSII protein abundances, an effect that was shown to be reversible upon Mn re-supplementation. Specifically, the extrinsic proteins PsbP and PsbQ showed Mn-dependent changes in abundances. Similar trends in the response to Mn deficiency at the protein level were observed when comparing DDA, SRM and LA-ICP-MS results. A biologically important exception to this trend was the loss of PsbO in the SRM analysis, which highlights the necessity of validating protein changes by more than one technique. The developed method enables a higher number of proteins to be multiplexed in comparison to existing immunoassays. Furthermore, multiplexed protein analysis by LA-ICP-MS provides an analytical platform with high throughput appropriate for screening large collections of plants.

Toll-like receptor 2-mediated alternative activation of microglia is protective after spinal cord injury.

Improving neurological outcome after spinal cord injury is a major clinical challenge because axons, once severed, do not regenerate but 'dieback' from the lesion site. Although microglia, the immunocompetent cells of the brain and spinal cord respond rapidly to spinal cord injury, their role in subsequent injury or repair remains unclear. To assess the role of microglia in spinal cord white matter injury we used time-lapse two-photon and spectral confocal imaging of green fluorescent protein-labelled microglia, yellow fluorescent protein-labelled axons, and Nile Red-labelled myelin of living murine spinal cord and revealed dynamic changes in white matter elements after laser-induced spinal cord injury in real time. Importantly, our model of acute axonal injury closely mimics the axonopathy described in well-characterized clinically relevant models of spinal cord injury including contusive-, compressive- and transection-based models. Time-lapse recordings revealed that microglia were associated with some acute pathophysiological changes in axons and myelin acutely after laser-induced spinal cord injury. These pathophysiological changes included myelin and axonal spheroid formation, spectral shifts in Nile Red emission spectra in axonal endbulbs detected with spectral microscopy, and 'bystander' degeneration of axons that survived the initial injury, but then succumbed to secondary degeneration. Surprisingly, modulation of microglial-mediated release of neurotoxic molecules failed to protect axons and myelin. In contrast, sterile stimulation of microglia with the specific toll-like receptor 2 agonist Pam2CSK4 robustly increased the microglial response to ablation, reduced secondary degeneration of central myelinated fibres, and induced an alternative (mixed M1:M2) microglial activation profile. Conversely, Tlr2 knock out: Thy1 yellow fluorescent protein double transgenic mice experienced greater axonal dieback than littermate controls. Thus, promoting an alternative microglial response through Pam2CSK4 treatment is neuroprotective acutely following laser-induced spinal cord injury. Therefore, anti-inflammatory treatments that target microglial activation may be counterintuitive after spinal cord injury.

MALDI mass spectrometry imaging in microscope mode with infrared lasers: bypassing the diffraction limits.

This letter demonstrates the use of infrared matrix-assisted laser desorption/ionization coupled with microscope mode mass spectrometry imaging. It is aimed to explore the use of intrinsic water in tissue as a matrix for imaging at spatial resolutions below the diffraction limit of the employed IR optics. Stigmatic ion optics with a magnification factor of ~70 were used to project the spatial distribution of produced ions onto a detector while separating ions with different mass-to-charge ratios using a time-of-flight mass spectrometer. A pixelated detector was used to simultaneously record arrival time and impact position. A previously described dried-droplet sample system of 2,5-dihydroxybenzoic acid (DHB) and 5 peptides covered by a copper grid for defined surface structure was used to benchmark the light- and ion-optical setup for spatial resolution and mass spectrometric performance. A spatial resolving power of 9.8 µm, well below the optical limit of diffraction (14 µm for the given setup), was established. After, frozen cryo-sections from a biological model system were measured by exploiting the endogenous water content as a matrix. Principal component analysis enabled a clear distinction between distinct tissue regions identified by both light microscopy and MS imaging.

Laser-printing of toner-based 96-microzone plates for immunoassays.

This work describes the quick and simple fabrication of toner-based 96-microzone plates by a direct-printing technology. The printer deposits a toner layer (ca. 5 µm thick) on the polyester surface which acts as a hydrophobic barrier to confine small volumes of sample on test zones (wells). A 96-microzone toner plate was explored to demonstrate its capability of performing enzyme-linked immunosorbent assay (ELISA). The detection of anti-immunoglobulin G (anti-IgG) and immunoglobulin M (IgM) antibodies has been successfully achieved in cell culture and serum samples, respectively. The use of a conventional microplate reader has allowed obtaining a limit of detection of 13 fmol of mouse IgG per zone on printed microplates. The IgM antibody has been detected in a serum sample collected from a patient infected with dengue virus. The detection of a primary infection has been provided by a microplate reader and also by a cell phone camera. Besides the bioanalytical feasibility, toner-based zones have shown good repeatability for inter-zone and intra-plate comparisons. The relative standard deviation (RSD) values for inter-zone (n = 12) and intra-plate (n = 3) comparisons were lower than 6% and 11%, respectively. Furthermore, it was found that the lifetime of each printed microplate depends on the storage temperature. The shelf life for devices stored at 10 °C has been estimated to be ca. four weeks.

Laser scanning as a useful tool in implant retrieval analysis: a demonstration using rotating platform and fixed bearing tibial inserts.

Objective methods for analyzing arthroplasty retrieval implants are needed. To address this, we used a readily available laser scanner to analyze damage deviations between cohorts of rotating platform and fixed bearing inserts previously analyzed using traditional, subjective retrieval analysis methods. We asked the following research questions: 1) Do articular surface deviations measured by the scanner correlate with the subjective damage scores? 2) Do articular surface deviations differ between inserts due to design differences? Correlations between deviations and damage scores were present in RP but not FB inserts. Seven different deviation patterns were present between the RP and FB inserts and were a function of design. In conclusion laser scanning was found to be a useful objective tool for analyzing arthroplasty retrievals.

In situ imaging of honeybee (Apis mellifera) venom components from aqueous and aluminum hydroxide-adsorbed venom immunotherapy preparations.

BACKGROUND: Treatment with aqueous and aluminum hydroxide (Al[OH](3))-adsorbed purified honeybee (Apis mellifera) venom (HBV) preparations can reduce the incidence of side effects associated with venom immunotherapy. OBJECTIVE: The aim of the present study was to assess these purified HBV immunotherapy preparations in situ. METHODS: Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) was used to visualize the distribution of HBV components. The preparations were administered on the back legs of naive Wistar rats. The rats were killed, and cryosectioned tissue sections were subjected to hematoxylin and eosin staining and MALDI-MSI analyses. RESULTS: Low-density maps of tissue distribution of HBV peptides, such as secapin, mast cell degranulating peptide, and melittin (Api m 4) were detected in the tissue after administration of HBV immunotherapy preparations. In addition, release of biogenic amines, cytokines, and leukotrienes was observed, and the distribution of HBV allergens, such as Api m 1 and Api m 2, was shown. At the 24-hour time point, the major HBV allergen Api m 1 was still detected at the site of Al(OH)(3)-adsorbed HVB injection, whereas in the case of aqueous HBV preparation, all the allergens, as well as most of the biogenic amines, were cleared at the 24-hour time point. CONCLUSION: The present study shows that the majority of low-molecular-weight HBV components are rapidly removed from the site of venom immunotherapy administration. Furthermore, Al(OH)(3)-adsorbed HBV preparation demonstrated a depot effect, prolonging the availability of bee venom allergens at the site of administration.

Salmon lice survive the straight shooter: A commercial scale sea cage trial of laser delousing.

Ectoparasitic salmon louse (Lepeophtheirus salmonis) infestations are costly for Atlantic salmon (Salmo salar) farmers in Norway. As a result, there is a strong desire for solutions to prevent and control infestations, and new technologies are typically developed and commercialised rapidly, without rigorous validation. Here, we tested the efficacy of a new commercially available control measure-delousing by underwater lasers-using a replicated design at full commercial scale. Laser delousing was used in combination with a preventive method (snorkel cages), with laser nodes deployed in 3 of the 6 sea cages at the site. The trial ran for 54 days, after which time there was no difference in infestation density of mobile salmon louse stages (pre-adult, adult male or adult female) in cages with or without laser nodes installed. By the end of the trial, adult female lice numbers in all cages were close to the legislated trigger for mandatory delousing (0.5 adult female lice per fish). The laser nodes delivered a large number of pulses relative to the number of lice in the cages, indicating that a lack of lethality rather than a lack of target detection was the limiting factor. If all pulses had been effective, they should have removed between 4-38 % of mobile lice each day. There was no effect on salmon welfare indicators such as skin condition or eye status. Our results highlight the importance of rigorous validation of new technologies across a range of conditions before widespread implementation by industry.

Facile analysis of metabolites by capillary electrophoresis coupled to matrix-assisted laser desorption/ionization mass spectrometry using target plates with polysilazane nanocoating and grooves.

We present an inexpensive method and apparatus for the deposition of analytes separated by capillary electrophoresis (CE) onto a custom-designed matrix-assisted laser desorption/ionization (MALDI) target plate. This dedicated CE-MALDI plate is coated with an omniphobic polysilazane nanocoating and has an array of parallel grooves acting as recipients of the separation effluent. The 3-D pattern in the top layer of the coated plate greatly improves loading of the matrix solution prior to separation and facilitates deposition of the separated species. We demonstrate application of this straightforward protocol to the analysis of metabolites from the central metabolic pathway in a complex biological sample spiked with small molecule standards.

[Analysis and experimental verification of sensitivity and SNR of laser warning receiver].

In order to countermeasure increasingly serious threat from hostile laser in modern war, it is urgent to do research on laser warning technology and system, and the sensitivity and signal to noise ratio (SNR) are two important performance parameters in laser warning system. In the present paper, based on the signal statistical detection theory, a method for calculation of the sensitivity and SNR in coherent detection laser warning receiver (LWR) has been proposed. Firstly, the probabilities of the laser signal and receiver noise were analyzed. Secondly, based on the threshold detection theory and Neyman-Pearson criteria, the signal current equation was established by introducing detection probability factor and false alarm rate factor, then, the mathematical expressions of sensitivity and SNR were deduced. Finally, by using method, the sensitivity and SNR of the sinusoidal grating laser warning receiver developed by our group were analyzed, and the theoretic calculation and experimental results indicate that the SNR analysis method is feasible, and can be used in performance analysis of LWR.

[Laser information technologies in oncosurgery].

The introduction of laser information technologies to medical practice has improved the results of reconstructive and plastic operations in oncology thanks to a possibility to plan the surgery beforehand. Sixty-eight patients were operated on at Gertsen Moscow Research Oncology Institute between 2001 and 2006; 99 stereolithographic models were used to plan different stages of the surgical intervention. The tasks fulfilled with an individual stereolithographic model, may be relatively divided into three groups: 1) planning the resection stage; 2) planning the reconstructive surgery; 3) assessing the necessity to correct the complex autograft after the reconstructive surgery. The use of laser stereolithographic modelling in surgical treatment of patients with locally advanced malignancies made it possible to complete the reconstructive operation successfully in 98.4% of the patients, and rehabilitate functionally 88.6% of the patients.

Fine welding with lasers.

The need for micro joining metallic alloys for surgical instruments, implants and advanced medical devices is driving a rapid increase in the implementation of laser welding technology in research, development and volume production. This article discusses the advantages of this welding method and the types of lasers used in the process.

Bulk and Thin Film Synthesis of Compositionally Variant Entropy-stabilized Oxides.

Here, we present a procedure for the synthesis of bulk and thin film multicomponent (Mg0.25(1-x)CoxNi0.25(1-x)Cu0.25(1-x)Zn0.25(1-x))O (Co variant) and (Mg0.25(1-x)Co0.25(1-x)Ni0.25(1-x)CuxZn0.25(1-x))O (Cu variant) entropy-stabilized oxides. Phase pure and chemically homogeneous (Mg0.25(1-x)CoxNi0.25(1-x)Cu0.25(1-x)Zn0.25(1-x))O (x = 0.20, 0.27, 0.33) and (Mg0.25(1-x)Co0.25(1-x)Ni0.25(1-x)CuxZn0.25(1-x))O (x = 0.11, 0.27) ceramic pellets are synthesized and used in the deposition of ultra-high quality, phase pure, single crystalline thin films of the target stoichiometry. A detailed methodology for the deposition of smooth, chemically homogeneous, entropy-stabilized oxide thin films by pulsed laser deposition on (001)-oriented MgO substrates is described. The phase and crystallinity of bulk and thin film materials are confirmed using X-ray diffraction. Composition and chemical homogeneity are confirmed by X-ray photoelectron spectroscopy and energy dispersive X-ray spectroscopy. The surface topography of thin films is measured with scanning probe microscopy. The synthesis of high quality, single crystalline, entropy-stabilized oxide thin films enables the study of interface, size, strain, and disorder effects on the properties in this new class of highly disordered oxide materials.

Experimental Methods for Spin- and Angle-Resolved Photoemission Spectroscopy Combined with Polarization-Variable Laser.

The goal of this protocol is to present how to perform spin- and angle-resolved photoemission spectroscopy combined with polarization-variable 7-eV laser (laser-SARPES), and demonstrate a power of this technique for studying solid state physics. Laser-SARPES achieves two great capabilities. Firstly, by examining orbital selection rule of linearly polarized lasers, orbital selective excitation can be carried out in SAPRES experiment. Secondly, the technique can show full information of a variation of the spin quantum axis as a function of the light polarization. To demonstrate the power of the collaboration of these capabilities in laser-SARPES, we apply this technique for the investigations of spin-orbit coupled surface states of Bi2Se3. This technique affords to decompose spin and orbital components from the spin-orbit coupled wavefunctions. Moreover, as a representative advantage of using the direct spin detection collaborated with the polarization-variable laser, the technique unambiguously visualizes the light polarization dependence of the spin quantum axis in three-dimension. Laser-SARPES dramatically increases a capability of photoemission technique.

Custom Engineered Tissue Culture Molds from Laser-etched Masters.

As the field of tissue engineering has continued to mature, there has been increased interest in a wide range of tissue parameters, including tissue shape. Manipulating tissue shape on the micrometer to centimeter scale can direct cell alignment, alter effective mechanical properties, and address limitations related to nutrient diffusion. In addition, the vessel in which a tissue is prepared can impart mechanical constraints on the tissue, resulting in stress fields that can further influence both the cell and matrix structure. Shaped tissues with highly reproducible dimensions also have utility for in vitro assays in which sample dimensions are critical, such as whole tissue mechanical analysis. This manuscript describes an alternative fabrication method utilizing negative master molds prepared from laser etched acrylic: these molds perform well with polydimethylsiloxane (PDMS), permit designs with dimensions on the centimeter scale and feature sizes smaller than 25 µm, and can be rapidly designed and fabricated at a low cost and with minimal expertise. The minimal time and cost requirements allow for laser etched molds to be rapidly iterated upon until an optimal design is determined, and to be easily adapted to suit any assay of interest, including those beyond the field of tissue engineering.

Continuous-wave Thulium Laser for Heating Cultured Cells to Investigate Cellular Thermal Effects.

An original method to heat cultured cells using a 1.94 µm continuous-wave thulium laser for biological assessment is introduced here. Thulium laser radiation is strongly absorbed by water, and the cells at the bottom of the culture dish are heated through thermal diffusion. A laser fiber with a diameter of 365 µm is set about 12 cm above the culture dish, without any optics, such that the laser beam diameter is almost equivalent to the inner diameter of the culture dish (30 mm). By keeping a consistent amount of culture medium in each experiment, it is possible to irradiate the cells with a highly reproducible temperature increase. To calibrate the temperature increase and its distribution in one cell culture dish for each power setting, the temperature was measured during 10 s of irradiation at different positions and at the cellular level. The temperature distribution was represented using a mathematical graphics software program, and its pattern across the culture dish was in Gaussian form. After laser irradiation, different biological experiments could be performed to assess temperature-dependent cell responses. In this manuscript, viability staining (i.e., distinguishing live, apoptotic, and dead cells) is introduced to help determine the threshold temperatures for cell apoptosis and death after different points in time. The advantages of this method are the preciseness of the temperature and the time of heating, as well as its high efficiency in heating cells in a whole cell culture dish. Furthermore, it allows for study with a wide variety of temperatures and time durations, which can be well-controlled by a computerized operating system.