Dual stereo-digital image correlation system for simultaneous measurement of overlapped wings with a polarization RGB camera and fluorescent speckle patterns.

Simultaneous monitoring of overlapped multi-wing structure by stereo-digital image correlation (stereo-DIC) may be used to quantify insect motion and deformation. We propose a dual stereo-DIC system based on multispectral imaging with a polarization RGB camera. Different fluorescent speckle patterns were fabricated on wings, which emit red and blue spectra under ultraviolet light that were imaged and separated using a polarization RGB camera and auxiliary optical splitting components. The resulting dual stereo-DIC system was validated through translation experiments with transparent sheets and reconstructed overlapped insect wings (cicadas). Dynamic measurements of the Ruban artificial flier indicate the efficacy of this approach to determining real insect flight behavior.

Full spectrum fluorescence lifetime imaging with 0.5 nm spectral and 50 ps temporal resolution.

The use of optical techniques to interrogate wide ranging samples from semiconductors to biological tissue for rapid analysis and diagnostics has gained wide adoption over the past decades. The desire to collect ever more spatially, spectrally and temporally detailed optical signatures for sample characterization has specifically driven a sharp rise in new optical microscopy technologies. Here we present a high-speed optical scanning microscope capable of capturing time resolved images across 512 spectral and 32 time channels in a single acquisition with the potential for ~0.2 frames per second (256 × 256 image pixels). Each pixel in the resulting images contains a detailed data cube for the study of diverse time resolved light driven phenomena. This is enabled by integration of system control electronics and on-chip processing which overcomes the challenges presented by high data volume and low imaging speed, often bottlenecks in previous systems.

Random lasers from photonic crystal wings of butterfly and moth for speckle-free imaging.

Several biological membranes have been served as scattering materials of random lasers, but few of them include natural photonic crystals. Here, we propose and demonstrate a facile approach to fabricating high-performance biological photonic crystal random lasers, which is cost-effective and reproducible for mass production. As a benchmark, optical and lasing properties of dye-coated Lepidoptera wings, including Papilio ulysses butterfly and Chrysiridia rhipheus moth, are characterized and show a stable laser emission with a superior threshold of 0.016 mJ/cm2, as compared to previous studies. To deploy the proposed devices in practical implementation, we have applied the as-fabricated biological devices to bright speckle-free imaging applications, which is a more sustainable and more accessible imaging strategy.

Functional Analysis of ESCRT-Positive Extracellular Vesicles in the Drosophila Wing Imaginal Disc.

A large number of studies have shown that proteins of the Endosomal Sorting Complex Required for Transport (ESCRT) can trigger the biogenesis of different types of Extracellular Vesicles (EV). The functions that these vesicular carriers exert in vivo remain, however, poorly understood. In this chapter, we describe a series of experimental approaches that we established in the Drosophila wing imaginal disc to study the importance of ESCRT-positive EVs for the extracellular transport of signaling molecules, as exemplified by a functional analysis of the mechanism of secretion and propagation of the major developmental morphogen Hedgehog (Hh).Through the combined use of genetic, cell biological, and imaging approaches, we investigate four important aspects of exovesicle biology: (1) The genetic identification of ESCRT proteins that are specifically required for Hh secretion. (2) The imaging of ESCRT and Hh-positive EVs in the lumenal space of both living and fixed wing imaginal discs. (3) The receptor-mediated capture of Hh-containing EVs on the surface of Hh-receiving cells. (4) The effect of manipulations of ESCRT function on the extracellular pool of Hh ligands.

Adult Drosophila muscle morphometry through microCT reveals dynamics during ageing.

Indirect flight muscles (IFMs) in adult Drosophila provide the key power stroke for wing beating. They also serve as a valuable model for studying muscle development. An age-dependent decline in Drosophila free flight has been documented, but its relation to gross muscle structure has not yet been explored satisfactorily. Such analyses are impeded by conventional histological preparations and imaging techniques that limit exact morphometry of flight muscles. In this study, we employ microCT scanning on a tissue preparation that retains muscle morphology under homeostatic conditions. Focusing on a subset of IFMs called the dorsal longitudinal muscles (DLMs), we find that DLM volumes increase with age, partially due to the increased separation between myofibrillar fascicles, in a sex-dependent manner. We have uncovered and quantified asymmetry in the size of these muscles on either side of the longitudinal midline. Measurements of this resolution and scale make substantive studies that test the connection between form and function possible. We also demonstrate the application of this method to other insect species making it a valuable tool for histological analysis of insect biodiversity.

Unveiling the nonlinear optical response of Trictenotoma childreni longhorn beetle.

The wings of some insect species are known to fluoresce under illumination by ultraviolet light. Their fluorescence properties are however, not comprehensively documented. In this article, the optical properties of one specific insect, the Trictenotoma childreni yellow longhorn beetle, were investigated using both linear and nonlinear optical (NLO) methods, including one- and two-photon fluorescence and second harmonic generation (SHG). These three distinct optical signals discovered in this beetle are attributed to the presence of fluorophores embedded within the scales covering their elytra. Experimental evidence collected in this study indicates that the fluorophores are non-centrosymmetric, a fundamental requirement for SHG. This study is the first reported optical behavior of this type in insects. We described how NLO techniques can complement other more convenient approaches to achieve a more comprehensive understanding of insect scales and integument properties.

The peregrine falcon's rapid dive: on the adaptedness of the arm skeleton and shoulder girdle.

During a dive, peregrine falcons (Falco peregrinus) can reach a velocity of up to 320 km h- 1. Our computational fluid dynamics simulations show that the forces that pull on the wings of a diving peregrine can reach up to three times the falcon's body mass at a stoop velocity of 80 m s- 1 (288 km h- 1). Since the bones of the wings and the shoulder girdle of a diving peregrine falcon experience large mechanical forces, we investigated these bones. For comparison, we also investigated the corresponding bones in European kestrels (Falco tinnunculus), sparrow hawks (Accipiter nisus) and pigeons (Columba livia domestica). The normalized bone mass of the entire arm skeleton and the shoulder girdle (coracoid, scapula, furcula) was significantly higher in F. peregrinus than in the other three species investigated. The midshaft cross section of the humerus of F. peregrinus had the highest second moment of area. The mineral densities of the humerus, radius, ulna, and sternum were highest in F. peregrinus, indicating again a larger overall stability of these bones. Furthermore, the bones of the arm and shoulder girdle were strongest in peregrine falcons.

Dragonfly wing nodus: A one-way hinge contributing to the asymmetric wing deformation.

Dragonfly wings are highly specialized locomotor systems, which are formed by a combination of several structural components. The wing components, also known as structural elements, are responsible for the various aspects of the wing functionality. Considering the complex interactions between the wing components, modelling of the wings as a whole is only possible with inevitable huge oversimplifications. In order to overcome this difficulty, we have recently proposed a new approach to model individual components of complex wings comparatively. Here, we use this approach to study nodus, a structural element of dragonfly wings which has been less studied to date. Using a combination of several imaging techniques including scanning electron microscopy (SEM), wide-field fluorescence microscopy (WFM), confocal laser scanning microscopy (CLSM) and micro-computed tomography (micro-CT) scanning, we aim to characterize the spatial morphology and material composition of fore- and hindwing nodi of the dragonfly Brachythemis contaminata. The microscopy results show the presence of resilin in the nodi, which is expected to help the deformability of the wings. The computational results based on three-dimensional (3D) structural data suggest that the specific geometry of the nodus restrains its displacements when subjected to pressure on the ventral side. This effect, resulting from an interlocking mechanism, is expected to contribute to the dorso-ventral asymmetry of wing deformation and to provide a higher resistance to aerodynamic forces during the downstroke. Our results provide an important step towards better understanding of the structure-property-function relationship in dragonfly wings. STATEMENT OF SIGNIFICANCE: In this study, we investigate the wing nodus, a specialized wing component in dragonflies. Using a combination of modern imaging techniques, we demonstrate the presence of resilin in the nodus, which is expected to facilitate the wing deformability in flight. The specific geometry of the nodus, however, seems to restrain its displacements when subjected to pressure on the ventral side. This effect, resulting from an interlocking mechanism, is suggested to contribute to dorso-ventral asymmetry of wing deformations and to provide a higher resistance to aerodynamic forces during the downstroke. Our results provide an important step towards better understanding of the structure-property-function relationship in dragonfly wings and might help to design more efficient wings for biomimetic micro-air vehicles.

Use of a caudoventral-craniodorsal oblique radiographic view made at 45° to the frontal plane to evaluate the pectoral girdle in raptors.

OBJECTIVE: To evaluate use of a caudoventral-craniodorsal oblique radiographic view made at 45° to the frontal plane (H view) for assessment of the pectoral (thoracic) girdle in raptors. DESIGN: Retrospective cross-sectional analysis. ANIMALS: 24 raptors suspected to have a fracture of the thoracic girdle. PROCEDURES: Standard ventrodorsal and H views were obtained for all birds. Radiographs were evaluated twice by a radiologist blinded to the final diagnosis, with each view first evaluated independently and views then evaluated in combination. Sensitivity, specificity, positive predictive value, and negative predictive value were calculated, with results of surgery or necropsy used as the gold standard. RESULTS: 9 birds had thoracic girdle fractures; fractures were correctly identified in 8 of these 9 birds on the ventrodorsal view alone, 7 of these 9 birds on the H view alone, and all 9 birds on the 2 views in combination. Fifteen birds did not have thoracic girdle fractures; radiographs were correctly classified in 12 of these 15 birds when the ventrodorsal view was evaluated alone, all 15 birds when the H view was evaluated alone, and 14 of these 15 birds when the 2 views were evaluated in combination. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that the H view or the addition of the H view to the VD view could be useful in raptors suspected to have fractures of the thoracic girdle. Agreement with the gold standard (ie, fracture present or absent) was higher with the H view and combination of views than with the ventrodorsal view alone.

Ultrastructure of dragonfly wing veins: composite structure of fibrous material supplemented by resilin.

Dragonflies count among the most skilful of the flying insects. Their exceptional aerodynamic performance has been the subject of various studies. Morphological and kinematic investigations have showed that dragonfly wings, though being rather stiff, are able to undergo passive deformation during flight, thereby improving the aerodynamic performance. Resilin, a rubber-like protein, has been suggested to be a key component in insect wing flexibility and deformation in response to aerodynamic loads, and has been reported in various arthropod locomotor systems. It has already been found in wing vein joints, connecting longitudinal veins to cross veins, and was shown to endow the dragonfly wing with chordwise flexibility, thereby most likely influencing the dragonfly's flight performance. The present study revealed that resilin is not only present in wing vein joints, but also in the internal cuticle layers of veins in wings of Sympetrum vulgatum (SV) and Matrona basilaris basilaris (MBB). Combined with other structural features of wing veins, such as number and thickness of cuticle layers, material composition, and cross-sectional shape, resilin most probably has an effect on the vein's material properties and the degree of elastic deformations. In order to elucidate the wing vein ultrastructure and the exact localisation of resilin in the internal layers of the vein cuticle, the approaches of bright-field light microscopy, wide-field fluorescence microscopy, confocal laser-scanning microscopy, scanning electron microscopy and transmission electron microscopy were combined. Wing veins were shown to consist of up to six different cuticle layers and a single row of underlying epidermal cells. In wing veins of MBB, the latter are densely packed with light-scattering spheres, previously shown to produce structural colours in the form of quasiordered arrays. Longitudinal and cross veins differ significantly in relative thickness of exo- and endocuticle, with cross veins showing a much thicker exocuticle. The presence of resilin in the unsclerotised endocuticle suggests its contribution to an increased energy storage and material flexibility, thus to the prevention of vein damage. This is especially important in the highly stressed longitudinal veins, which have much lower possibility to yield to applied loads with the aid of vein joints, as the cross veins do. These results may be relevant not only for biologists, but may also contribute to optimise the design of micro-air vehicles.

Computational Aerodynamic Analysis of a Micro-CT Based Bio-Realistic Fruit Fly Wing.

The aerodynamic features of a bio-realistic 3D fruit fly wing in steady state (snapshot) flight conditions were analyzed numerically. The wing geometry was created from high resolution micro-computed tomography (micro-CT) of the fruit fly Drosophila virilis. Computational fluid dynamics (CFD) analyses of the wing were conducted at ultra-low Reynolds numbers ranging from 71 to 200, and at angles of attack ranging from -10° to +30°. It was found that in the 3D bio-realistic model, the corrugations of the wing created localized circulation regions in the flow field, most notably at higher angles of attack near the wing tip. Analyses of a simplified flat wing geometry showed higher lift to drag performance values for any given angle of attack at these Reynolds numbers, though very similar performance is noted at -10°. Results have indicated that the simplified flat wing can successfully be used to approximate high-level properties such as aerodynamic coefficients and overall performance trends as well as large flow-field structures. However, local pressure peaks and near-wing flow features induced by the corrugations are unable to be replicated by the simple wing. We therefore recommend that accurate 3D bio-realistic geometries be used when modelling insect wings where such information is useful.

Kinematics of chiropteran shoulder girdle in flight.

New data on the mechanisms of movements of the shoulder girdle and humerus of bats are described; potential mobility is compared to the movements actually used in flight. The study was performed on the basis of morphological and functional analysis of anatomical specimens of 15 species, high speed and high definition filming of two species and X-ray survey of Rousettus aegyptiacus flight. Our observations indicate that any excursions of the shoulder girdle in bats have relatively small input in the wing amplitude. Shoulder girdle movements resemble kinematics of a crank mechanism: clavicle plays the role of crank, and scapula-the role of connecting rod. Previously described osseous "locking mechanisms" in shoulder joint of advanced bats do not affect the movements, actually used in flight. The wing beats in bats are performed predominantly by movements of humerus relative to shoulder girdle, although these movements occupy the caudal-most sector of available shoulder mobility.

Bi-functional photonic structure in the Papilio nireus (Papilionidae): modeling by scattering-matrix optical simulations.

Scales of the Papilio nireus combine fluorophores confined in a natural photonic structure. By means of numerical simulations based on the scattering-matrix formalism, we reveal the bi-functional optical role of this peculiar architecture. Two aspects are considered: the absorption of an incident light flux and the emission of another luminous flux. First, results highlight a light trapping effect and a light absorption increase in the ultraviolet, visible and near infrared ranges. Then, results highlight an enhanced fluorescence occurring in the spatial as well as in the frequency domain. This observation could be of great interest to design new optical devices.

Vascular contrast enhanced micro-CT imaging of "radiators" in the Brazilian free-tailed bat (Tadarida brasiliensis).

The Brazilian free-tailed bat (Tadarida brasiliensis) exhibits a highly vascularized, hairless thermal window (or "radiator") on the proximal ventral surfaces of extended wings and body. We identified this character using thermal infrared imaging and investigated the vasculature using barium sulfate enhanced microcomputed tomography (micro-CT). Micro-CT images revealed unique arrangements of arteries and veins in the region of the radiator positioned perpendicular to the axis of the body. Coupling micro-CT imaging with analysis of surface temperature profiles, we concluded that radiators aid in thermoregulation during flight in variable environments. This study represents the first application of contrast enhanced micro-CT to visualize vasculature of bats and thus exhibits a promising technique for further investigations of cardiovascular function and anatomy in bats.

Morphological and kinematic basis of the hummingbird flight stroke: scaling of flight muscle transmission ratio.

Hummingbirds (Trochilidae) are widely known for their insect-like flight strokes characterized by high wing beat frequency, small muscle strains and a highly supinated wing orientation during upstroke that allows for lift production in both halves of the stroke cycle. Here, we show that hummingbirds achieve these functional traits within the limits imposed by a vertebrate endoskeleton and muscle physiology by accentuating a wing inversion mechanism found in other birds and using long-axis rotational movement of the humerus. In hummingbirds, long-axis rotation of the humerus creates additional wing translational movement, supplementing that produced by the humeral elevation and depression movements of a typical avian flight stroke. This adaptation increases the wing-to-muscle-transmission ratio, and is emblematic of a widespread scaling trend among flying animals whereby wing-to-muscle-transmission ratio varies inversely with mass, allowing animals of vastly different sizes to accommodate aerodynamic, biomechanical and physiological constraints on muscle-powered flapping flight.

X-ray imaging with the PILATUS 100k detector.

We report on the application of the PILATUS 100K pixel detector for medical imaging. Experimental results are presented in the form of X-ray radiographs using standard X-ray absorption contrast and a recently developed phase contrast imaging method. The results obtained with the PILATUS detector are compared to results obtained with a conventional X-ray imaging system consisting of an X-ray scintillation screen, lens optics, and a charge coupled device. Finally, the results for both systems are discussed more quantitatively based on an image power spectrum analysis.

Congenital skeletal abnormalities in a tawny owl chick (Strix aluco).

In this case report, we describe a tawny owl chick (Strix aluco) coming from a Wild Fauna Recovery Center with multiple congenital malformations in the limbs. The animal was unable to fly and showed marked malnutrition and poor general appearance. Physical, radiologic, and anatomic examinations showed osseous malformations including dislocation of radius and carpometacarpus with abnormal nonfunctional fixation of ligamentum propatagialis, absence of most parts of the bones of the manus in both wings, and twisted nonfused left tarsometatarsus with marked griphosis of digits. Routine toxicologic and pathologic examinations did not reveal a specific etiology.

Lithium fluoride as a novel X-ray image detector for biological mu-world capture.

X-ray microradiographs of small biological objects, such as animals and plant materials at micrometric resolution, are currently performed by various methods, all of which are limited by the resolution or the dynamic range of the image detectors. Here a novel X-ray image detector is discussed, in which the previous limitations have been overcome. A film of lithium fluoride salt is used as a detector, in which the stored biological image is read by observing the optically stimulated visible luminescence of the active color centers, efficiently produced by the X-rays.

Acoustic microscopic analysis of the biological structure of insect wing membranes with emphasis on their waxy surface.

The mechanical performance of natural materials depends on the type, and especially the composition of the molecular constituents. They are almost without exception composite materials, whose characteristics are determined by the characteristics of the individual constituents, their shape, their interaction, and in particular their orientation within the natural material. One of the most impressive natural composites is the insect cuticle. This lightweight building material impresses one with its ability to withstand extremely heavy loading. Even the ultrathin (3-10 microm) membranes of insect wings add greatly to the structural stability of the wings. By means of acoustic microscopy, the present study also shows that the thin covering of wax on the membrane is not an accidental material arrangement. Contrary to that of locust wings, dragonfly wing membranes were found to have a criss-cross fiber-like density gradient within to the waxy layer. This density gradient proved to be mechanically relevant in stabilizing the wings.