Manipulation and growth of birefringent protein crystals in optical tweezers

We report on the trapping, rotation, and in-situ growth of birefringent tetragonal lysozyme crystals in optical tweezers operating at a wavelength of 1070 nm. Variation of the pH and lysozyme concentration of the solution during growth was used to alter the length to width ratio of the crystals, and hence their orientation in the tweezers. Crystals with the optical axis skewed or perpendicular to the trapping-beam axis could be rotated by changing the orientation of linearly polarized light. We observed spontaneous spinning of some asymmetric crystals in the presence of linearly polarized light, due to radiation pressure effects. Addition of protein to the solution in the tweezers permitted real-time observation of crystal growth. (C) 2004 Optical Society of America.

Information Modelling of Two-Dimensional Optical Parameters Measurement

A method for measurement and visualization of the complex transmission coefficient of 2-D micro- objects is proposed. The method is based on calculation of the transmission coefficient from the diffraction pattern and the illumination aperture function for monochromatic light. A phase-stepping method was used for diffracted light phase determination.

Searching for and positioning of contextualized learning objects

Learning object economies are marketplaces for the sharing and reuse of learning objects (LO). There are many motivations for stimulating the development of the LO economy. The main reason is the possibility of providing the right content, at the right time, to the right learner according to adequate quality standards in the context of a lifelong learning process; in fact, this is also the main objective of education. However, some barriers to the development of a LO economy, such as the granularity and editability of LO, must be overcome. Furthermore, some enablers, such as learning design generation and standards usage, must be promoted in order to enhance LO economy. For this article, we introduced the integration of distributed learning object repositories (DLOR) as sources of LO that could be placed in adaptive learning designs to assist teachers’ design work. Two main issues presented as a result: how to access distributed LO, and where to place the LO in the learning design. To address these issues, we introduced two processes: LORSE, a distributed LO searching process, and LOOK, a micro context-based positioning process, respectively. Using these processes, the teachers were able to reuse LO from different sources to semi-automatically generate an adaptive learning design without leaving their virtual environment. A layered evaluation yielded good results for the process of placing learning objects from controlled learning object repositories into a learning design, and permitting educators to define different open issues that must be covered when they use uncontrolled learning object repositories for this purpose. We verified the satisfaction users had with our solution

Using objects to coordinate design activity in interaction

The coordination of design is a multi-faceted problem in construction. In design interactions in particular the real-time coordination of design activity is a persistent concern. The use of objects to coordinate the activity of design is studied as this happens in interactions between an architect and a building user group, in a setting where maintaining awareness of the design situation is important. An account of ways in which this was accomplished and how design activity is coordinated through interactional practices is provided. The empirical analyses examine design interaction from an ethnomethodological/conversation analysis (EM/CA) informed perspective to examine: ways in which mutual orientation to design issues are accomplished, how objects can provide a resource for the recognition of the activities of others and ways in which objects might be observable as momentarily intelligible. Subtle interactional practices involving talk, gesture and gaze were some of the small ways in which mutual orientation to the design actions of others became observable. The production of actions sequentially, in response to another's action, marked the real-time coordination of design moves in this setting. The relevance of accounts of micro-interaction to develop understanding of design activity and how it is coordinated are considered.

A palaeobiologist's guide to 'virtual' micro-CT preparation

This paper provides a brief but comprehensive guide to creating, preparing and dissecting a 'virtual' fossil, using a worked example to demonstrate some standard data processing techniques. Computed tomography (CT) is a 3D imaging modality for producing 'virtual' models of an object on a computer. In the last decade, CT technology has greatly improved, allowing bigger and denser objects to be scanned increasingly rapidly. The technique has now reached a stage where systems can facilitate large-scale, non-destructive comparative studies of extinct fossils and their living relatives. Consequently the main limiting factor in CT-based analyses is no longer scanning, but the hurdles of data processing (see disclaimer). The latter comprises the techniques required to convert a 3D CT volume (stack of digital slices) into a virtual image of the fossil that can be prepared (separated) from the matrix and 'dissected' into its anatomical parts. This technique can be applied to specimens or part of specimens embedded in the rock matrix that until now have been otherwise impossible to visualise. This paper presents a suggested workflow explaining the steps required, using as example a fossil tooth of Sphenacanthus hybodoides (Egerton), a shark from the Late Carboniferous of England. The original NHMUK copyrighted CT slice stack can be downloaded for practice of the described techniques, which include segmentation, rendering, movie animation, stereo-anaglyphy, data storage and dissemination. Fragile, rare specimens and type materials in university and museum collections can therefore be virtually processed for a variety of purposes, including virtual loans, website illustrations, publications and digital collections. Micro-CT and other 3D imaging techniques are increasingly utilized to facilitate data sharing among scientists and on education and outreach projects. Hence there is the potential to usher in a new era of global scientific collaboration and public communication using specimens in museum collections.

Molecular motors-based micro- and nano-biocomputation devices

Protein molecular motors, which are natural nano-machines that convert the chemical energy into mechanical work for cellular motion, muscle contraction and cell division, have been integrated in the last decade in primitive nanodevices based on the motility of nano-biological objects in micro- and nano-fabricated structures. However, the motility of microorganisms powered by molecular motors has not been similarly exploited. Moreover, among the proposed devices based on molecular motors, i.e., nanosensors, nano-mechanical devices and nano-imaging devices, biocomputation devices are conspicuously missing. The present contribution discusses, based on the present state of the art nano- and micro-fabrication, the comparative advantages and disadvantages of using nano- and micro-biological objects in future computation devices. (c) 2006 Elsevier B.V. All rights reserved.

Spectral analysis of four 'hypervariable' AGN: a micro-needle in the haystack?

We analyze four extreme AGN transients to explore the possibility that they are caused by rare, high-amplitude microlensing events. These previously unknown type-I AGN are located in the redshift range 0.6-1.1 and show changes of > 1.5 magnitudes in the g-band on a timescale of ~years. Multi-epoch optical spectroscopy, from the William Herschel Telescope, shows clear differential variability in the broad line fluxes with respect to the continuum changes and also evolution in the line profiles. In two cases a simple point-source, point-lens microlensing model provides an excellent match to the long-term variability seen in these objects. For both models the parameter constraints are consistent with the microlensing being due to an intervening stellar mass object but as yet there is no confirmation of the presence of an intervening galaxy. The models predict a peak amplification of 10.3/13.5 and an Einstein timescale of 7.5/10.8 years respectively. In one case the data also allow constraints on the size of the CIII] emitting region, with some simplifying assumptions, to to be ~1.0-6.5 light-days and a lower limit on the size of the MgII emitting region to be > 9 light-days (half-light radii). This CIII] radius is perhaps surprisingly small. In the remaining two objects there is spectroscopic evidence for an intervening absorber but the extra structure seen in the lightcurves requires a more complex lensing scenario to adequately explain.

Noninvasive imaging of three-dimensional micro and nanostructures by topological methods

We present topological derivative and energy based procedures for the imaging of micro and nano structures using one beam of visible light of a single wavelength. Objects with diameters as small as 10 nm can be located and their position tracked with nanometer precision. Multiple objects dis-tributed either on planes perpendicular to the incidence direction or along axial lines in the incidence direction are distinguishable. More precisely, the shape and size of plane sections perpendicular to the incidence direction can be clearly determined, even for asymmetric and nonconvex scatterers. Axial resolution improves as the size of the objects decreases. Initial reconstructions may proceed by gluing together two-dimensional horizontal slices between axial peaks or by locating objects at three-dimensional peaks of topological energies, depending on the effective wavenumber. Below a threshold size, topological derivative based iterative schemes improve initial predictions of the lo-cation, size, and shape of objects by postprocessing fixed measured data. For larger sizes, tracking the peaks of topological energy fields that average information from additional incident light beams seems to be more effective.

Ihwg-μnir: A Miniaturised Near-infrared Gas Sensor Based On Substrate-integrated Hollow Waveguides Coupled To A Micro-nir-spectrophotometer.

A miniaturised gas analyser is described and evaluated based on the use of a substrate-integrated hollow waveguide (iHWG) coupled to a microsized near-infrared spectrophotometer comprising a linear variable filter and an array of InGaAs detectors. This gas sensing system was applied to analyse surrogate samples of natural fuel gas containing methane, ethane, propane and butane, quantified by using multivariate regression models based on partial least square (PLS) algorithms and Savitzky-Golay 1(st) derivative data preprocessing. The external validation of the obtained models reveals root mean square errors of prediction of 0.37, 0.36, 0.67 and 0.37% (v/v), for methane, ethane, propane and butane, respectively. The developed sensing system provides particularly rapid response times upon composition changes of the gaseous sample (approximately 2 s) due the minute volume of the iHWG-based measurement cell. The sensing system developed in this study is fully portable with a hand-held sized analyser footprint, and thus ideally suited for field analysis. Last but not least, the obtained results corroborate the potential of NIR-iHWG analysers for monitoring the quality of natural gas and petrochemical gaseous products.

Estudo por espectroscopia micro-Raman dos mecanismos de separação de fase em vidros fosfatos de metais de transição

Glass-ceramics are prepared by controlled separation of crystal phases in glasses, leading to uniform and dense grain structures. On the other hand, chemical leaching of soluble crystal phases yields porous glass-ceramics with important applications. Here, glass/ceramic interfaces of niobo-, vanado- and titano-phosphate glasses were studied by micro-Raman spectroscopy, whose spatial resolution revealed the multiphase structures. Phase-separation mechanisms were also determined by this technique, revealing that interface composition remained unchanged as the crystallization front advanced for niobo- and vanadophosphate glasses (interface-controlled crystallization). For titanophosphate glasses, phase composition changed continuously with time up to the equilibrium composition, indicating a spinodal-type phase separation.

Determinação espectrofotométrica de hexametilenotetramina (HMT) em medicamentos, utilizando ácido cromotrópico e forno de micro-ondas

This paper proposes a methodology for spectrophotometric determination of hexamethylenetetramine (HMT) by using chromotropic acid in a phosphoric acid media employing a domestic microwave oven as a source of heating. The reddish-purple soluble product is quantitatively formed after 30 s of irradiation and obeys the Beer´s law in the range between 0.1-1.2 mg L-1 HMT (r = 0.99925). The method was applied successfully in commercial pharmaceutical preparations containing dyes in their composition. The results showed that the method proposed is feasible for simplicity, speed, low cost, precision and accuracy when compared with United States Pharmacopeia official method.

The Rg1 allele as a valuable tool for genetic transformation of the tomato 'Micro-Tom' model system

Background: The cultivar Micro-Tom (MT) is regarded as a model system for tomato genetics due to its short life cycle and miniature size. However, efforts to improve tomato genetic transformation have led to protocols dependent on the costly hormone zeatin, combined with an excessive number of steps. Results: Here we report the development of a MT near-isogenic genotype harboring the allele Rg1 (MT-Rg1), which greatly improves tomato in vitro regeneration. Regeneration was further improved in MT by including a two-day incubation of cotyledonary explants onto medium containing 0.4 mu M 1-naphthaleneacetic acid (NAA) before cytokinin treatment. Both strategies allowed the use of 5 mu M 6-benzylaminopurine (BAP), a cytokinin 100 times less expensive than zeatin. The use of MT-Rg1 and NAA pre-incubation, followed by BAP regeneration, resulted in high transformation frequencies (near 40%), in a shorter protocol with fewer steps, spanning approximately 40 days from Agrobacterium infection to transgenic plant acclimatization. Conclusions: The genetic resource and the protocol presented here represent invaluable tools for routine gene expression manipulation and high throughput functional genomics by insertional mutagenesis in tomato.

Analysis of the tip roundness effects on the micro- and macroindentation response of elastic-plastic materials

In this work, the effects of indenter tip roundness oil the load-depth indentation curves were analyzed using finite element modeling. The tip roundness level was Studied based on the ratio between tip radius and maximum penetration depth (R/h(max)), which varied from 0.02 to 1. The proportional Curvature constant (C), the exponent of depth during loading (alpha), the initial unloading slope (S), the correction factor (beta), the level of piling-up or sinking-in (h(c)/h(max)), and the ratio h(max)/h(f) are shown to be strongly influenced by the ratio R/h(max). The hardness (H) was found to be independent of R/h(max) in the range studied. The Oliver and Pharr method was successful in following the variation of h(c)/h(max) with the ratio R/h(max) through the variation of S with the ratio R/h(max). However, this work confirmed the differences between the hardness values calculated using the Oliver-Pharr method and those obtained directly from finite element calculations; differences which derive from the error in area calculation that Occurs when given combinations of indented material properties are present. The ratio of plastic work to total work (W(p)/W(t)) was found to be independent of the ratio R/h(max), which demonstrates that the methods for the Calculation of mechanical properties based on the *indentation energy are potentially not Susceptible to errors caused by tip roundness.

Importance of considering a material micro-failure criterion in the numerical modelling of the shot peening process applied to parabolic leaf springs

Shot peening is a cold-working mechanical process in which a shot stream is propelled against a component surface. Its purpose is to introduce compressive residual stresses on component surfaces for increasing the fatigue resistance. This process is widely applied in springs due to the cyclical loads requirements. This paper presents a numerical modelling of shot peening process using the finite element method. The results are compared with experimental measurements of the residual stresses, obtained by the X-rays diffraction technique, in leaf springs submitted to this process. Furthermore, the results are compared with empirical and numerical correlations developed by other authors.

Convergence of developmental mutants into a single tomato model system: 'Micro-Tom' as an effective toolkit for plant development research

Background: The tomato (Solanum lycopersicum L.) plant is both an economically important food crop and an ideal dicot model to investigate various physiological phenomena not possible in Arabidopsis thaliana. Due to the great diversity of tomato cultivars used by the research community, it is often difficult to reliably compare phenotypes. The lack of tomato developmental mutants in a single genetic background prevents the stacking of mutations to facilitate analysis of double and multiple mutants, often required for elucidating developmental pathways. Results: We took advantage of the small size and rapid life cycle of the tomato cultivar Micro-Tom (MT) to create near-isogenic lines (NILs) by introgressing a suite of hormonal and photomorphogenetic mutations (altered sensitivity or endogenous levels of auxin, ethylene, abscisic acid, gibberellin, brassinosteroid, and light response) into this genetic background. To demonstrate the usefulness of this collection, we compared developmental traits between the produced NILs. All expected mutant phenotypes were expressed in the NILs. We also created NILs harboring the wild type alleles for dwarf, self-pruning and uniform fruit, which are mutations characteristic of MT. This amplified both the applications of the mutant collection presented here and of MT as a genetic model system. Conclusions: The community resource presented here is a useful toolkit for plant research, particularly for future studies in plant development, which will require the simultaneous observation of the effect of various hormones, signaling pathways and crosstalk.