Confocal laser scanning microscopy elucidation of the micromorphology of the leaf cuticle and analysis of its chemical composition

Electron microscopy techniques such as transmission electron microscopy (TEM) and scanning electron microscopy (SEM) have been invaluable tools for the study of the micromorphology of plant cuticles. However, for electron microscopy, the preparation techniques required may invariably introduce artefacts in cuticle preservation. Further, there are a limited number of methods available for quantifying the image data obtained through electron microscopy. Therefore, in this study, optical microscopy techniques were coupled with staining procedures and, along with SEM were used to qualitatively and quantitatively assess the ultrastructure of plant leaf cuticles. Leaf cryosections of Triticum aestivum (wheat), Zea mays (maize), and Lupinus angustifolius (lupin) were stained with either fat-soluble azo stain Sudan IV or fluorescent, diarylmethane Auramine O and were observed under confocal laser scanning microscope (CLSM). For all the plant species tested, the cuticle on the leaf surfaces could be clearly resolved in many cases into cuticular proper (CP), external cuticular layer (ECL), and internal cuticular layer (ICL). Novel image data analysis procedures for quantifying the epicuticular wax micromorphology were developed, and epicuticular waxes of L. angustifolius were described here for the first time. Together, application of a multifaceted approach involving the use of a range of techniques to study the plant cuticle has led to a better understanding of cuticular structure and provides new insights into leaf surface architecture.

Ranking of fibre-reinforced composite plate surface finish quality by wavelet texture analysis

In the automotive and other industries, the visual appearance of external surfaces is a key factor in perceived product quality. Traditionally, the quality of an automotive surface finish has been judged by expert human auditors. A set of 17 fibre-reinforced composite plates was previously manufactured to have a range of surface finish qualities and these plates were ranked by three expert observers and also optically digitally imaged. Following validation of the previous rankings, the wavelet texture analysis (WTA) technique was applied to the digital photographs to derive an instrumental measure of surface finish quality based on the panel images. The rank correlation between the human expert surface finish quality ratings and those from the W TA image analysis process was found to be positive, large and statistically significant. This finding indicates that WTA could form the basis of an inexpensive and practical instrumental method for the ranking of fibre-reinforced composite surface finish quality.

Mapping land use in a large agricultural basin : a comparison between classification techniques

In order to facilitate the better management of river basin resources, the Glenelg-Hopkins region in south-east Australia required an accurate and up to date land use map. Land use has a major impact on Australia's natural resources including its soil, water, flora and fauna and plays a major role in determining basin health. Inappropriate land use and practices have contributed to extensive dryland salinity and water quality problems. Land use data is often required for environmental models and in most cases the reliability of model outputs is dependent on the spatial detail and accuracy of the land use mapping. This paper examines methods to obtain an up to date land use map and a detailed accuracy assessment using Landsat ETM+ data for a regional basin. A multi-source based approach allowed the collection of 4817 ground truth data points from the field investigation. This enabled researchers to (i) incorporate a full range of information into digital image analysis with significant improvements in accuracy and (ii) hold sufficient independent references for an accurate error assessment. Classification accuracy was significantly improved using a stratification design, in which the region is sub-divided into smaller homogenous areas as opposed to a full scene classification technique. The overall classification accuracy was 84% (KHAT= 0.833) for the stratified approach compared to 76% (KHAT= 0.743) for the full scene classification. Effective assessment, planning and management of basins are dependent on a sound knowledge of the distribution and variability of land use.

Objective classification of fabric pilling based on the two-dimensional discrete wavelet transform

A number of methods for automated objective ratings of fabric pilling based on image analysis are described in the literature. The periodic structure of fabrics makes them suitable candidates for frequency domain analysis. We propose a new method of frequency domain analysis based on the two-dimensional discrete wavelet transform to objectively measure pilling intensity in sample images. We present a preliminary evaluation of the proposed method based on analysis of two series of standard pilling evaluation test images. The initial results suggest that the proposed method is feasible, and that the ability of the method to discriminate between levels of pilling intensity depends on the wavelet analysis scale being closely matched to the fabric interyarn pitch. We also present a heuristic method for optimal selection of an analysis wavelet and associated analysis scale.

Evaluation of the fineness of degummed bast fibers

Fiber fineness characteristics are important for yarn production and quality. In this paper, degummed bast fibers such as hemp, flax and ramie have been examined with the Optical Fiber Diameter Analyzer (OFDA100 and OFDA2000) systems for fiber fineness, in comparison with the conventional image analysis and the Wira airflow tester. The correlation between the results from these measurements was analysed. The results indicate that there is a significant linear co-relation between the fiber fineness measurement results obtained from those different systems. In addition, the mean fiber width and its coefficient of variation obtained from the OFDA100 system are smaller than those obtained from the OFDA2000 system, due to the difference in sample preparation methods. The OFDA2000 system can also measure the fiber fineness profile along the bast fiber plants, which can be useful for plant breeding.

Characterization and application of objective pilling classification to patterned fabrics

Previously, the authors proposed a new, simple method of frequency domain analysis based on the two-dimensional discrete wavelet transform to objectively measure the pilling intensity in sample fabric images. The method was further characterized, and the results obtained indicate that standard deviation and variance are the most appropriate measures of the dispersion of wavelet details coefficients for analysis, that the relationship between wavelet analysis scale and fabric inter-yarn pitch was empirically confirmed, and, that fabrics with random patterns do not appear to impact on the effectiveness of the analysis method.

The determination of the local conditions for void initiation in front of a crack tip for materials with second-phase particles

A procedure is proposed to determine, for second-phase particles near a crack tip, the maximum particle stresses at the moment of void initiation by either particle fracture or particle/matrix interface separation. A digital image analysis system is applied to perform a quantitative analysis of corresponding fracture surface regions from stereo image pairs taken in the scanning electron microscope. The fracture surface analysis is used to measure, for individual particles, the crack tip opening displacement at the moment of void initiation and the particle location with respect to the crack tip. From these data, the stress tensor at the moment of void initiation is calculated from the Hutchinson–Rice–Rosengren (HRR) field theory. The corresponding average local stresses within the particle are evaluated by a non-linear Mori–Tanaka-type approach. These stresses are compared to estimates according to the models by Argon et al. [A.S. Argon, J. Im, R. Safoglu, Metall. Trans. 6 (1975) 825] and Beremin [F.M. Beremin, Metall. Trans. 12 (1981) 723]. The procedure is demonstrated on an Al6061–10% Al2O3 metal matrix composite.

Objective assessment of pilling of nonwoven fabrics using the two dimensional discrete wavelet transform

Fabric pilling is a serious problem for the apparel industry, causing an unsightly appearance and premature wear. Woolen products are particularly prone to pilling. Recently, a process for production of woolen nonwoven apparel fabrics has been commercialized in Australia, and may lead to new markets for Australian wool. However, the success of such nonwoven fabrics will partly rely on their propensity to pill. A key element in the control of fabric pilling is the evaluation of resistance to pilling by testing. Resistance to pilling is normally tested in the laboratory by processes that simulate accelerated wear, followed by a manual assessment of the degree of pilling by an expert based on a visual comparison of the sample to a set of test images. To bring more objectivity into the pilling rating process, a number of automated systems based on image analysis have been developed. The authors previously proposed a new method of image analysis based on the two-dimensional discrete wavelet transform to objectively measure the pilling intensity for woven fabrics. This paper presents preliminary work in extending this method to nonwoven fabrics.

Increased versican content is associated with tendinosis pathology in the patellar tendon of athletes with jumper

Expansion of the extracellular matrix is a prominent but poorly characterized feature of tendinosis. The present study aimed to characterize the extent and distribution of the large aggregating proteoglycan versican in patients with patellar tendinosis. We obtained tendon from tendinopathy patients undergoing debridement of the patellar tendon and from controls undergoing intramedullary tibial nailing. Versican content was investigated by Western blotting and immunohistochemistry. Microvessel thickness and density were determined using computer-assisted image analysis. Markers for smooth muscle actin, endothelial cells (CD31) and proliferating cells (Ki67) were examined immunohistochemically. Western blot analysis and immunohistochemical staining revealed elevated versican content in the proximal patellar tendon of tendinosis patients (P=0.042). Versican content was enriched in regions of fibrocartilage metaplasia and fibroblast proliferation, as well as in the perivascular matrix of proliferating microvessels and within the media and intima of arterioles. Microvessel density was higher in tendinosis tissue compared with control tissue. Versican deposition is a prominent feature of patellar tendinosis. Because this molecule is not only a component of normal fibrocartilagenous matrices but also implicated in a variety of soft tissue pathologies, future studies should further detail both pathological and adaptive roles of versican in tendons.

Effect of alloying elements on the microstructure property relationship in thermomechanically processed C-Mn-Si TRIP steels

The effect of additions of Nb, Al and Mo to Fe-C-Mn-Si TRIP steel on the final microstructure and mechanical properties after simulated  thermomechanical processing (TMP) has been studied. The laboratory simulations of discontinuous cooling during TMP were performed using a hot rolling mill. All samples were characterised using optical microscopy and image analysis. The volume fraction of retained austenite was ascertained using a heat tinting technique and X-ray diffraction measurements. Room temperature mechanical properties were determined by a tensile test. From this a comprehensive understanding of the structural aspect of the bainite transformation in these types of TRIP steels has been developed. The  results have shown that the final microstructures of thermomechanically processed TRIP steels comprise 50 % of polygonal ferrite, 7 - 12 % of retained austenite, non-carbide bainitic structure and martensite. All steels exhibited a good combination of ultimate tensile strength and total elongation. The microstructure-property examination revealed the relationship between the composition of TRIP steels and their mechanical properties. It has been shown that the addition of Mo to the C-Si-Mn-Nb TRIP steel increases the ultimate tensile strength up to 1020 MPa. The stability of the retained austenite of the Nb-Mo steel was degraded, which led to a decrease in the elongation (24 %). The results have demonstrated that the addition of Al to C-Si-Mn-Nb steel leads to a good combination of strength (∼ 940 MPa) and elongation (∼ 30 %) due to the formation of refined acicular ferrite and granular bainite structure with ∼7 - 8 % of stable retained austenite. Furthermore, it has been found that the addition of Al increases the volume fraction of bainitic ferrite laths. The investigations have shown an interesting result that, in the Nb-Mo-Al steel, Al has a more pronounced effect on the microstructure in comparison with Mo. It has been found that the bainitic structure of the Nb-Mo-Al steel appears to be more granular than in the Nb-Mo steel. Moreover, the volume fraction of the retained austenite increased (12 %) with decreasing bainitic ferrite content. The results have demonstrated that this steel has the best mechanical properties (1100 MPa and 28 % elongation). It has been concluded that the combined effect of Nb, Mo, and Al addition on the dispersion of the bainite, martensite and retained austenite in the ferrite matrix and the morphology of these phases is different than effect of Nb, Mo and Al, separately.

Correlation between optical and SEM measurements of wool cortical cell size

Wool fibres consist of micro to nano scale protein constituents that could be used for innovative applications. While techniques for extracting these constituents or making wool fibres into organic powders have been developed, effectively dispersing the particles and accurately determining their size has been difficult in practice. In this study, an ultrasonic method was employed to disperse cortical cells extracted from wool fibres into an
immersion oil or ethanol. Specimens of the cortical cells were then observed under optical microscopy and scanning electron microscopy, respectively. Cell length and maximum cell diameter were measured to quantify the cell size. The results suggest significant discrepancies exist in the cortical cell size obtained from the two different measurement techniques. The maximum diameter of wool cortical cells obtained from the optical microscope was much larger than that from the scanning electron microscope, while the length was much shorter. A correction factor is given so that cortical cell size obtained from the two measurement techniques can be compared.

Effect of alloying elements on the microstructure-property relationship in thermomechanically processed C-Mn-Si TRIP steels

The effect of additions of Nb, Al and Mo to Fe-C-Mn-Si TRIP steel on the final microstructure and mechanical properties after simulated thermomechanical processing (TMP) has been studied. The laboratory simulations of discontinuous cooling during TMP were performed using a hot rolling mill. All samples were characterised using optical microscopy and image analysis. The volume fraction of retained austenite was ascertained using a heat tinting technique and X-ray diffraction measurements. Room temperature mechanical properties were determined by a tensile test. From this a comprehensive understanding of the structural aspect of the bainite transformation in these types of TRIP steels has been developed. The results have shown that the final microstructures of thermomechanically processed TRIP steels comprise 50 % of polygonal ferrite, 7 - 12 % of retained austenite, non-carbide bainitic structure and martensite. All steels exhibited a good combination of ultimate tensile strength and total elongation. The microstructure-property examination revealed the relationship between the composition of TRIP steels and their mechanical properties. It has been shown that the addition of Mo to the C-Si-Mn-Nb TRIP steel increases the ultimate tensile strength up to 1020 MPa. The stability of the retained austenite of the Nb-Mo steel was degraded, which led to a decrease in the elongation (24 %). The results have demonstrated that the addition of Al to C-Si-Mn-Nb steel leads to a good combination of strength (∼ 940 MPa) and elongation (∼ 30 %) due to the formation of refined acicular ferrite and granular bainite structure with ∼7 - 8 % of stable retained austenite. Furthermore, it has been found that the addition of Al increases the volume fraction of bainitic ferrite laths. The investigations have shown an interesting result that, in the Nb-Mo-Al steel, Al has a more pronounced effect on the microstructure in comparison with Mo. It has been found that the bainitic structure of the Nb-Mo-Al steel appears to be more granular than in the Nb-Mo steel. Moreover, the volume fraction of the retained austenite increased (12 %) with decreasing bainitic ferrite content. The results have demonstrated that this steel has the best mechanical properties (1100 MPa and 28 % elongation). It has been concluded that the combined effect of Nb, Mo, and Al addition on the dispersion of the bainite, martensite and retained austenite in the ferrite matrix and the morphology of these phases is different than effect of Nb, Mo and Al, separately.

Mimicry and the eye of the beholder

Recent experiments (Dittrich et al. (Proc. R. Soc. Lond. B 251, 195 (1993))) suggest that pigeon perception of wasp mimicry by hoverflies is similar to that of humans and of computer-based image matching. However, the relations are nonlinear and may explain why some species are abundant despite their being poor mimics to the human eye. We suggest that these discrepancies between pigeon and human categorization may lie in the differences between avian and primate colour vision. As pigeon categorization and computer image analysis were both assessed by using colour slides designed for human vision, they lacked the natural colour information available to wild birds, in particular that from ultraviolet (uv) wavelengths.