Combination Of Light Microscopy Research Articles

Sea-ice eukaryotes of the Gulf of Finland, Baltic Sea, and evidence for herbivory on weakly shade-adapted ice algae

To determine community composition and physiological status of early spring sea-ice organisms, we collected sea-ice, slush and under-ice water samples from the Baltic Sea. We combined light microscopy, HPLC pigment analysis and pyrosequencing, and related the biomass and physiological status of sea-ice algae with the protistan community composition in a new way in the area. In terms of biomass, centric diatoms including a distinct Melosira arctica bloom in the upper intermediate section of the fast ice, dinoflagellates, euglenoids and the cyanobacterium Aphanizomenon sp. predominated in the sea-ice sections and unidentified flagellates in the slush. Based on pigment analyses, the ice-algal communities showed no adjusted photosynthetic pigment pools throughout the sea ice, and the bottom-ice communities were not shade-adapted. The sea ice included more characteristic phototrophic taxa (49%) than did slush (18%) and under-ice water (37%). Cercozoans and ciliates were the richest taxon groups, and the differences among the communities arose mainly from the various phagotrophic protistan taxa inhabiting the communities. The presence of pheophytin a coincided with an elevated ciliate biomass and read abundance in the drift ice and with a high Eurytemora affinis read abundance in the pack ice, indicating that ciliates and Eurytemora affinis were grazing on algae.

The first instar larva of two species of Miltogramminae (Diptera: Sarcophagidae) from the Middle East – examples of peculiar morphology

AbstractThe first instar larva is described for two species of Miltogramminae flesh flies representing the genera Miltogramma Meigen and Phrosinella Robineau‐Desvoidy. Using a combination of light microscopy and scanning electron microscopy, the habitus is documented along with all important exoskeletal and cephaloskeletal structures. The described first instar larvae show substantial modifications of their morphology in comparison with the saprophagous ground plan defined for calyptrate flies. Miltogramma (Miltogrammidium) efflatouni Rohdendorf possesses the general set of derived character states shared with other species of Miltogramma and shares larval synapomorphies with species representing the subgenera Anacanthothecum Rohdendorf, Cylindrothecum Rohdendorf and Miltogrammidium Rohdendorf. Unique character states of the first instar larva of this species are the spine‐like shape of antennal complex, laterally compressed apical part of mouthhooks and an extreme elongation of the sensilla of the maxillary palpus. Phrosinella (Asiometopa) ujgura (Rohdendorf) possesses a pair of retractable, robust processes on the ventral margin of the first thoracic segment and a very strong, elongated labrum. Both character states are present in other known first instar larvae of genus Phrosinella. A reduction of cuticular ridges on dorsal and lateral surfaces of abdominal segments supports a monophylum consisting of the subgenera Asiometopa Rohdendorf and Caspiomyia Rohdendorf within genus Phrosinella.

SIMS and NanoSIMS analyses of well-preserved microfossils imply oxygen-producing photosynthesis in the Mesoproterozoic anoxic ocean

Well-preserved microfossils in the stromatolitic cherts from the Gaoyuzhuang Formation (~1500Ma), which have one of the best state of preservation in the Proterozoic Era, may play key roles in systematizing information about the evolution of early life in the Proterozoic Ocean. Here, a combination of light microscopy (LM), scanning electron microscopy (SEM), nano-scale secondary ion mass spectrometry (NanoSIMS), secondary ion mass spectrometry (SIMS) and geochemical techniques was employed to characterize the morphology, elemental distributions and carbon isotope values of individual Gaoyuzhuang microfossils, and their growing environment. Light microscopy analyses show that abundant filamentous and coccoid microfossils, classified as oscillatoriacean and chroococcacean cyanobacteria, respectively, are exceptionally well preserved in chert. NanoSIMS analyses show that metabolically important elements, such as 12C−, 13C−, 12C14N−, 32S−, and 34S−, are concentrated in these microfossils and that the variations in the concentrations of these elements are similar to each other, establishing the elemental patterns in undoubtedly biogenic microstructures. The carbon isotope (δ13C) values of individual microfossils range from −31.7‰±0.9‰ to −23.1‰±1.0‰ (weighted mean=−28.5‰±0.1‰), consistent not only with a Calvin Cycle-based photosynthesis, but typically for chroococcacean cyanobacteria as well. Our results, for the first time, provide the element distributions and cell specific carbon isotope values on convincing Mesoproterozoic cyanobacterial fossils, supporting sustained oxygen-producing photosynthesis in the Proterozoic Ocean. The geochemical data indicate these fossil microorganisms may grow in an anoxic seawater, potentially supporting the delayed oxygenation on the Earth. In this regard, this integrated approach may be a step towards a more comprehensive picture of the evolution of early life on the Earth.

The middle Miocene palynoflora and palaeoenvironments of Eskihisar (Yatağan basin, south-western Anatolia): a combined LM and SEM investigation

Anatolia was a crossroads for mammal migration during the Miocene due to intermittent land connections between Africa and Anatolia and persisting warm conditions. Here, we investigated a palynological section from middle Miocene sediments of Eskihisar (southwestern Anatolia) in order to establish biogeographic links of the palynoflora and to infer the palaeoenvironment. Four algal palynomorphs, nine spore taxa, eight gymnosperms, three monocots, and 67 dicot pollen types were encountered and investigated using the “single grain method” that combines light microscopy and scanning electron microscopy. Two pollen zones reflect different phases of basin development. Zonal vegetation remained fairly stable across the section and reflects heterogeneous environments including broad-leaved deciduous forest, subtropical forest, and sclerophyllous and semi-evergreen oak forest. Conifers were accessory elements in the broad-leaved deciduous forest communities and replaced these at higher elevations. Some herbaceous taxa (Plumbaginaceae) indicate scattered occurrences of sandy and/or rocky soils. Biogeographic affinities are general Northern Hemispheric, North American, and East Asian as also suggested by the macro fossil record. Only two taxa provide potential biogeographic links with the African flora. This suggests that biome shifts of plant taxa between African subtropical /tropical biomes and Anatolian (western Eurasian) temperate forests and shrublands may have been rare in the middle Miocene.

Morphological and molecular description of immature stages of Ornithodoros savignyi (Acari: Argasidae).

This study was designed to provide more details about larva, first nymph, and second nymph of Ornithodoros savignyi using a combination of light microscopy (LM), scanning electron microscopy (SEM), and partial sequence of mitochondrial 16S ribosomal ribonucleic acid (rRNA). The main characteristics of larva are wrinkled integument with many grooves, gnathosoma without camerostome cheeks, hypostome with a pair of large teeth apically, and tarsus without humps. The comparisons between the first and the second nymphs are different shape and distribution of dorsal grooves; a few spots without mammilla on the dorsal surface of the second nymph; 27 and 63-65 pairs of setae on the dorsal surface of the first and second nymphs, respectively; small holes on mammillae that are more dense in the second nymph; basis capitulum with two pairs of small setae in the second nymph; and one pair of sate in the first nymph, hypostome with dental formula 2/2 in the first nymph, and 3/3 in the second nymph. The partial 16S rRNA sequence of the second nymph that was determined as O. savignyi (450bp) was deposited in GenBank under the accession number KU163242.

Microscopy in combination with image analysis for characterization of fishmeal material in aquafeed

The aim of this study was to investigate the use of microscopy in combination with image analysis (IA) measurements for the characterization of fish bone lacunae in aquafeed-extracted material. For this purpose two experiments have been conducted.In experiment (exp.) 1, six samples of fish meal based aquafeed were analyzed by the microscopic method, according to Annex VI of Regulation 152/2009. Sediment fractions of each sample were observed with a compound microscope at ×40. Two hundred and fifty eight bone fragment lacunae images were recorded and processed through IA software. Accordingly, on each lacuna 30 geometric variables have been obtained and measured. The geometric variables have been grouped in two main families, namely size descriptors and derived shape descriptors.In exp. 2 measurements obtained from 1081 bone lacunae (644 for mammals and 437 for poultry) acquired from 14 mammalian and 7 poultry samples have been merged with the aquafeed dataset (258 bone lacunae). Results obtained in exp. 1 indicated that nearly two thirds of the descriptors presented differences among the analyzed samples. Differences in observed values were not systematically distributed among the six samples. Nevertheless, in all analyzed samples features of lacunae have shown an overlap. By contrast the comparison of fish bone lacunae with avian and mammalian bone lacunae (exp. 2), has revealed a large gap between terrestrial and aquatic animals in the case of several descriptors (16 primary and 8 secondary descriptors). Therefore, it can be concluded that combining light microscopy and image analysis: (i) fish material in aquafeed appears quite homogenous in term of bone features; (ii) fish material can be distinguished from avian and mammalian materials by selecting specific descriptors.

A combined light and scanning electron microscopy study

The uppermost Eocene Florissant Formation, Rocky Mountains, Colorado, has yielded numerous insect, vertebrate and plant fossils. Three previous comprehensive palynological studies investigated sections of lacustrine deposits of the Florissant Formation and documented the response of plant communities to volcanic eruptive phases but overall found little change in plant composition throughout the investigated sections. These studies reported up to 150 pollen and spore phenotypes. In the present paper, we used a taxonomic approach to the investigation of dispersed pollen and spores of the Florissant Formation. Sediment samples from the shale units containing macrofossils were investigated using light microscopy (LM) and scanning electron microscopy (SEM). The general picture of the palynoflora is in agreement with previous studies. However, the combined LM and SEM investigation provides important complementary information to previous LM studies. While a fairly large amount of previous pollen determinations could be confirmed, the purported taxonomic affinities of several pollen phenotypes need to be revised. For example, pollen referred to as Podocarpus or Podocarpidites sp. belongs to the Pinaceae Cathaya, Malus/Pyrus actually belongs to Dryadoideae, pollen of the form genus Boehlensipollis referred to as Proteaceae/Sapindaceae/Elaeagnaceae or Cardiospermum belongs to Sapindaceae but not to Cardiospermum, and pollen of Persicarioipollis sp. B with previously assumed affinities to Polygonaceae actually belongs to Thymelaeaceae. Pandaniidites and one type of Malvacipollis cannot be linked with Pandanaceae and Malvaceae. A few taxa are new records for Florissant (Ebenaceae: Diospyros; Mernispermaceae; Trochodendraceae: Tetracentron). In general, SEM investigations complement the LM palynological studies and improve the identification of dispersed pollen and spores and enable integration of data from dispersed fossil pollen into a wide range of comparative morphological, taxonomic, evolutionary, biogeographic and phylogenetic studies.

Stages of Cell Cannibalism--Entosis--in Normal Human Keratinocyte Culture.

Entosis is a type of cell cannibalism during which one cell penetrates into another cell and usually dies inside it. Researchers mainly pay attention to initial and final stages of entosis. Besides, tumor cells in suspension are the primary object of studies. In the present study, we investigated morphological changes of both cells-participants of entosis during this process. The substrate-dependent culture of human normal keratinocytes HaCaT was chosen for the work. A combination of light microscopy and scanning electron microscopy was used to prove that one cell was completely surrounded by the plasma membrane of another cell. We investigated such "cell-in-cell" structures and described the structural and functional changes of both cells during entosis. The outer cell nucleus localization and shape were changed. Gradual degradation of the inner cell nucleus and of the junctions between the inner and the outer cells was revealed. Moreover, repeated redistribution of the outer cell membrane organelles (Golgi apparatus, lysosomes, mitochondria, and autophagosomes), rearrangement of its cytoskeleton, and change in the lysosomal, autophagosomal, and mitochondrial state in both entotic cells were observed during entosis. On the basis of these data, we divided entosis into five stages that make it possible to systematize description of this type of cell death.

A simple and versatile cell wall staining protocol to study plant reproduction.

Key messageThe optical brightener SCRI Renaissance 2200 can be used as versatile dye to study various aspects of plant reproduction by confocal laser scanning microscopy. The study of sexual reproduction of plants has traditionally relied on light microscopy in combination with a variety of staining methods. Transgenic lines that label specific cell or tissue types with fluorescent proteins in combination with confocal laser scanning microscopy were an important development to visualize gametophyte development, the fertilization process, and to follow cell differentiation in the early embryo. Staining the cell perimeter to identify surrounding tissue is often a necessary prerequisite to put the fluorescent signal in the right context. Here, we present SCRI Renaissance 2200 (SR2200) as a versatile dye to study various aspects of plant reproduction ranging from pollen tube growth, guidance and reception to the early patterning process in the developing embryo of Arabidopsis thaliana. Furthermore, we demonstrate that SR2200 can be combined with a wide variety of fluorescent proteins. If spectral information can be recorded, even double labeling with dyes that have very similar emission spectra such as 4′,6-diamidin-2-phenylindol (DAPI) is possible. The presented staining method can be a single, easy-to-use alternative for a range of other staining protocols commonly used for microscopic analyses in plant reproductive biology.Electronic supplementary materialThe online version of this article (doi:10.1007/s00497-015-0267-1) contains supplementary material, which is available to authorized users.

Early postnatal diagnosis of hereditary spherocytosis by combining light microscopy, acidified glycerol lysis test and eosin-5'-maleimide binding assay.

Exact diagnosis of hereditary spherocytosis (HS) is widely considered unreliable around birth. However, early postnatal diagnosis at the beginning of congenital hemolysis may be essential for managing neonatal anemia and hemolytic icterus, identifying those at high risk for severe hyperbilirubinemia, irreversible kernicterus, or sudden need for red cell transfusion. We analyzed 37 blood samples from neonates or infants up to six weeks of life that had been collected in-house or shipped to our laboratory due to suspected red cell membrane disorder. By combining assessment of red cell morphology, acidified glycerol lysis test (AGLT), and eosin-5'-maleimide (EMA) binding assay, we were able to clearly exclude HS in 22 and confirm HS in 10 patients, of which one had undergone red cell transfusion prior to blood sampling. Assessment of red cell morphology and normal test results allowed diagnosis of infantile pyknocytosis or Heinz body anemia in three neonates. Re-evaluation of five patients with inconsistent results of AGLT and EMA binding led to confirmation of HS in two cases. Automated analysis of hematologic parameters revealed elevated proportion of hyperdense cells to be a highly significant indicator for HS in neonatal infants. We showed that assessment of red cell morphology in combination with AGLT and EMA binding assay is a reliable basis for confirming or rejecting suspected diagnosis of HS even in neonates. Our data underline the necessity for blood sampling and laboratory exploration in suspected red cell membrane or enzyme defects at the earliest occasion.

Morphotypes of Prorocentrum lima (Dinophyceae) from Hainan Island, South China Sea: morphological and molecular characterization

:The benthic dinoflagellate Prorocentrum lima has a cosmopolitan distribution that extends from temperate to tropical oceans. The identification of P. lima needs detailed morphological observation and molecular characterization because of its similarity to congeners and its morphological variation. In this study, we characterized different morphotypes of P. lima found in six sampling sites from Hainan Island, South China Sea, associated with macroalgae and seagrass. Twenty strains were isolated and cultivated, and P. lima was characterized using light and scanning electron microscopy in combination with large-subunit (LSU) and internal transcribed spacer (ITS) rDNA phylogeny. High morphological variability and genetic divergence was observed. Molecular analysis based on LSU and ITS rDNA sequences clearly indicated that the different morphotypes of P. lima were genetically based. The morphological and molecular data determined here suggest that all five morphotypes of P. lima examined from Hainan Island belonged to a P. lima complex.

The potential of bioorthogonal chemistry for correlative light and electron microscopy: a call to arms

With correlative light and electron microscopy (CLEM), the ultrastructural cellular location of a biomolecule of interest can be determined using a combination of light microscopy (LM) and electron microscopy (EM). In many cases, the application of CLEM requires the use of markers that need to be attached to a biomolecule of interest to allow its identification and localization. Here, we review the potential of bioorthogonal chemistry to introduce such markers for CLEM.

FLIPPER, a combinatorial probe for correlated live imaging and electron microscopy, allows identification and quantitative analysis of various cells and organelles.

Ultrastructural examination of cells and tissues by electron microscopy (EM) yields detailed information on subcellular structures. However, EM is typically restricted to small fields of view at high magnification; this makes quantifying events in multiple large-area sample sections extremely difficult. Even when combining light microscopy (LM) with EM (correlated LM and EM: CLEM) to find areas of interest, the labeling of molecules is still a challenge. We present a new genetically encoded probe for CLEM, named “FLIPPER”, which facilitates quantitative analysis of ultrastructural features in cells. FLIPPER consists of a fluorescent protein (cyan, green, orange, or red) for LM visualization, fused to a peroxidase allowing visualization of targets at the EM level. The use of FLIPPER is straightforward and because the module is completely genetically encoded, cells can be optimally prepared for EM examination. We use FLIPPER to quantify cellular morphology at the EM level in cells expressing a normal and disease-causing point-mutant cell-surface protein called EpCAM (epithelial cell adhesion molecule). The mutant protein is retained in the endoplasmic reticulum (ER) and could therefore alter ER function and morphology. To reveal possible ER alterations, cells were co-transfected with color-coded full-length or mutant EpCAM and a FLIPPER targeted to the ER. CLEM examination of the mixed cell population allowed color-based cell identification, followed by an unbiased quantitative analysis of the ER ultrastructure by EM. Thus, FLIPPER combines bright fluorescent proteins optimized for live imaging with high sensitivity for EM labeling, thereby representing a promising tool for CLEM.Electronic supplementary materialThe online version of this article (doi:10.1007/s00441-015-2142-7) contains supplementary material, which is available to authorized users.

Influence of cerium addition on microstructure and properties of Sn–Cu–(Ag) solder alloys

Abstract In the present work, the effect of Ce concentration on microstructure refinement, melting behavior and mechanical properties of a series of near-eutectic Sn–Ag–Cu alloys have been investigated. Twelve different alloys with Ce concentration between 0.005 and 0.171 wt% were prepared by melting of the pure elements in high purity argon, followed by quenching to room temperature. The samples microstructure and phase constitution were investigated by a combination of light microscopy, scanning electron microscopy, X-ray diffraction, and energy dispersive X-ray spectroscopy. In the alloys a total of five different phases have been identified: β-Sn, Cu 6 Sn 5 , Ag 3 Sn, CeO 2 , and CeSn 3 . The Ce addition is found to contribute to the refinement of mostly eutectic microstructures. The CeO 2 and/or CeSn 3 phases are observed in alloys with high Ce concentration. The absence of these particles in alloys with low Ce concentration (0.005–0.011 wt%) shows a complete dissolution of Ce in the β-Sn solid solution. The elongation and ultimate tensile strength of the alloys were determined under a constant strain rate of 2 mm min −1 . Moreover, the toughness (i.e. specific fracture energy) was calculated from the area underneath the force–extension curve. Melting temperatures are found to change slightly with bulk Ce concentration. Solution strengthening by Ce atoms, grain refinement, and the stress concentration along both types of Ce-rich particles of undesirable shapes are considered to be the main factors influencing the mechanical properties of the investigated alloy systems.

Current taxonomic studies on the diatom flora (Bacillariophyceae) of the Bolivian Altiplano, South America, with possible consequences for palaeoecological assessments

Abstract. The present investigation is based on detailed taxonomical analyses combining light microscopy (LM), scanning electron microscopy (SEM) and critical reviews of publications and type material. This method revealed a higher number of diatom taxa than that reported for the Altiplano, reports that are based on LM and SEM, but force-fitting the taxa into their European relatives. It also became evident that some taxa often reported from the Altiplano do not occur or are less common in this region and that misidentifications are the consequence not only of force-fitting, but also of misinterpretation of the original protologues and illustrations (taxonomic concept drift). These findings have important repercussions on past palaeoecological assessments for the Altiplano, which have been based on ecological information for taxa developing in ecosystems of highly contrasting conditions. Here we present the case of three araphid diatoms, which were reported as known taxa from Europe and elsewhere, but in fact correspond to taxa described as new for the Altiplano or unpublished taxa. It is recommended that: (1) a flora for this region is developed utilizing sound microscopical data and reviewing pertinent type material, (2) key common taxa used for past palaeoecological studies in the Altiplano are reviewed using a taxonomically thorough and critical method, and (3) future palaeoecological studies for the Andes are supported by prior detailed taxonomical analysis.

Correlative light and electron microscopy imaging of autophagy in a zebrafish infection model

High-resolution imaging of autophagy has been used intensively in cell culture studies, but so far it has been difficult to visualize this process in detail in whole animal models. In this study we present a versatile method for high-resolution imaging of microbial infection in zebrafish larvae by injecting pathogens into the tail fin. This allows visualization of autophagic compartments by light and electron microscopy, which makes it possible to correlate images acquired by the 2 techniques. Using this method we have studied the autophagy response against Mycobacterium marinum infection. We show that mycobacteria during the progress of infection are frequently associated with GFP-Lc3-positive vesicles, and that 2 types of GFP-Lc3-positive vesicles were observed. The majority of these vesicles were approximately 1 μm in size and in close vicinity of bacteria, and a smaller number of GFP-Lc3-positive vesicles was larger in size and were observed to contain bacteria. Quantitative data showed that these larger vesicles occurred significantly more in leukocytes than in other cell types, and that approximately 70% of these vesicles were positive for a lysosomal marker. Using electron microscopy, it was found that approximately 5% of intracellular bacteria were present in autophagic vacuoles and that the remaining intracellular bacteria were present in phagosomes, lysosomes, free inside the cytoplasm or occurred as large aggregates. Based on correlation of light and electron microscopy images, it was shown that GFP-Lc3-positive vesicles displayed autophagic morphology. This study provides a new approach for injection of pathogens into the tail fin, which allows combined light and electron microscopy imaging in vivo and opens new research directions for studying autophagy process related to infectious diseases.

New Method for (Nano)Micro-Characterization of Materials - Correlative Microscopy

New building materials must meet strict requirements on mechanical properties. Therefore structural and chemical characterization of new building materials is very important. The correlative microscopy is a new possibility of materials analysis. This method combines light microscopy (LM) and scanning electron microscopy (SEM). LM images can be then precisely extended by their morphological background and/or material distribution, e.g. with energy dispersive spectroscopy (EDS). With using correlative microscopy, the weakest places of alkali-activated concrete were found.

Evolutionary trends and ecological differentiation in early Cenozoic Fagaceae of western North America.

• The early Cenozoic was a key period of evolutionary radiation in Fagaceae. The common notion is that species thriving in the modern summer-dry climate of California originated in climates with ample summer rain during the Paleogene.• We investigated in situ and dispersed pollen of Fagaceae from the uppermost Eocene Florissant fossil beds, Colorado, United States, using a combined light and scanning electron microscopy approach.• Pollen types of Castaneoideae with affinities to modern Castanea, Lithocarpus, and Castanopsis were recognized. Pollen of the extinct genus Fagopsis represents a derived type of Castaneoideae pollen. Infrageneric groups of Quercus were well represented, including pollen of Group Protobalanus. The taxonomic diversity of Fagaceae and of the total plant assemblage indicates a mosaic of microclimates, that range from pronounced to weakly seasonal climates and depend on slope aspect and elevation. Continental climatic conditions may have triggered the evolution of sclerophyllous leaves and adaptive radiation in Quercus and other taxa thriving today under distinctly summer-dry and winter-dry climates.• Vegetation types similar to modern vegetation belts of the Coastal Ranges (chaparral, nemoral conifer forest) were established in the Front Range in the late Eocene. Coeval plant assemblages from the Coastal Ranges of California indicate distinctly subtropical, moist climates. Hence, characteristic elements found today in the summer-dry and winter-dry climates of Pacific North America (Quercus Group Protobalanus, Notholithocarpus) may opportunistically have dispersed into their modern ranges later in the Cenozoic. This scenario is in contrast to the evolution and migration patterns of their western Eurasian Mediterranean counterparts (Quercus Group Ilex).

Selective vulnerability of the cerebral vasculature to blast injury in a rat model of mild traumatic brain injury.

BackgroundBlast-related traumatic brain injury (TBI) is a common cause of injury in the military operations in Iraq and Afghanistan. How the primary blast wave affects the brain is not well understood. The aim of the present study was to examine whether blast exposure affects the cerebral vasculature in a rodent model. We analyzed the brains of rats exposed to single or multiple (three) 74.5 kPa blast exposures, conditions that mimic a mild TBI. Rats were sacrificed 24 hours or between 6 and 10 months after exposure. Blast-induced cerebral vascular pathology was examined by a combination of light microscopy, immunohistochemistry, and electron microscopy.ResultsWe describe a selective vascular pathology that is present acutely at 24 hours after injury. The vascular pathology is found at the margins of focal shear-related injuries that, as we previously showed, typically follow the patterns of penetrating cortical vessels. However, changes in the microvasculature extend beyond the margins of such lesions. Electron microscopy revealed that microvascular pathology is found in regions of the brain with an otherwise normal neuropil. This initial injury leads to chronic changes in the microvasculature that are still evident many months after the initial blast exposure.ConclusionsThese studies suggest that vascular pathology may be a central mechanism in the induction of chronic blast-related injury.

Cell differentiation and tissue formation in the unique fruits of devil's claws (Martyniaceae).

• Premise of the study: Martyniaceae are characterized by capsules with two upwardly curved, horn-shaped extensions representing morphologically specialized epizoochorous fruits. Because the capsules are assumed to cling to hooves and ankles of large mammals, fiber arrangement and tissue combinations within the endocarp ensuring proper attachment to the vector's feet during transport are of particular interest. In this first detailed anatomical investigation, the functional adaptation of the fruits and their implications for the specific dispersal mode are provided. The peculiar fiber arrangement may also be of interest for future biomimetic composite materials.• Methods: Endocarp anatomy and details of tissue differentiation were examined in fruits of Ibicella lutea and Proboscidea louisianica subsp. fragrans combining light microscopy, SEM, and x-ray microtomography analysis.• Key results: While tips of the extensions are predominantly reinforced by longitudinally oriented fibers, in the middle segment these fibers are densely packed in individual bundles entwined and separated by transversely elongated cells. Within the capsule wall, the fiber bundles are embedded in a dense mesh of transversely oriented fibers that circularly reinforce and protect the loculus. This fibrous pericarp tissue develops within few days by localized cell divisions and intrusive growth of primarily isodiametric parenchyma cells in the pistil.• Conclusions: The study allows insight into a unique and complex example of functionally driven cell growth and tissue formation. Long-horned fruits of Martyniaceae obviously are highly specialized to epizoochorous dispersal, pointing to primary vector-related seed dispersal. The highly ordered arrangement of fibers results in a great mechanical firmness.