bluuebunny/crossref_metadata_2025_split huggingface dataset:
Non‐optical bimorph‐based tapping‐mode force sensing method for scanning near‐field optical microscopy
Authors: SHANG G. Y. , LEI F. H. , TROYON M. , QIAO W. H. , TRUSSARDI‐REGNIER A. , MANFAIT M. 
Published: July 2004

SummaryA non‐optical bimorph‐based tapping‐mode force sensing method for tip–sample distance control in scanning near‐field optical microscopy is developed. Tapping‐mode force sensing is accomplished by use of a suitable piezoelectric bimorph cantilever, attaching an optical fibre tip to the extremity of the cantilever free end and fixing the guiding portion of the fibre to a stationary part near the tip to decouple it from the cantilever. This method is mainly characterized by the use of a bimorph, which carries out simultaneous excitation and detection of mechanical vibration at its resonance frequency owing to piezoelectric and anti‐piezoelectric effects, resulting in simplicity, compactness, ease of implementation and lack of parasitic optical background. In conjugation with a commercially available SPM controller, tapping‐mode images of various samples, such as gratings, human breast adenocarcinoma cells, red blood cells and a close‐packed layer of 220‐nm polystyrene spheres, have been obtained. Furthermore, topographic and near‐field optical images of a layer of polystyrene spheres have also been taken simultaneously. The results suggest that the tapping‐mode set‐up described here is reliable and sensitive, and shows promise for biological applications.
X‐ray high‐resolution vascular network imaging
Authors: PLOURABOUÉ F. , CLOETENS P. , FONTA C. , STEYER A. , LAUWERS F. , MARC‐VERGNES J.‐P. 
Published: July 2004

SummaryThis paper presents the first application of high‐resolution X‐ray synchrotron tomography to the imaging of large microvascular networks in biological tissue samples. This technique offers the opportunity of analysing the full three‐dimensional vascular network from the micrometre to the millimetre scale. This paper presents the specific sample preparation method and the X‐ray imaging procedure. Either barium or iron was injected as contrast agent in the vascular network. The impact of the composition and concentration of the injected solution on the X‐ray synchrotron tomography images has been studied. Two imaging modes, attenuation and phase contrast, are compared. Synchrotron high‐resolution computed tomography offers new prospects in the three‐dimensional imaging of in situ biological vascular networks.
Shape modelling of spatial particles from planar central sections – a case study
Authors: HANNILA M.‐L. , JENSEN E. B. VEDEL , HOBOLTH A. , NYENGAARD J. R. 
Published: July 2004

SummaryIn this paper we develop statistical tools for shape modelling of spatial particles from central sections through the particles. The particles are assumed to be star‐shaped with respect to a reference point inside the particles and are modelled as stochastic deformations of spheres centred at the reference points. The resulting particles are rotation invariant with respect to the reference point. As an illustration, the model is applied to study shape differences between neurons in the Granular and CA1 layer in the human hippocampus.
Quantitative fluorescence resonance energy transfer (FRET) measurement with acceptor photobleaching and spectral unmixing
Authors: GU Y. , DI W. L. , KELSELL D. P. , ZICHA D. 
Published: July 2004

SummaryFluorescence resonance energy transfer (FRET) by acceptor photobleaching is a simple but effective tool for measurements of protein–protein interactions. Until recently, it has been restricted to qualitative or relative assessments owing to the spectral bleed‐through contamination resulting from fluorescence overlap between the donor and the acceptor. In this paper, we report a quantitative algorithm that combines the spectral unmixing technique with FRET by acceptor photobleaching. By spectrally unmixing the emissions before and after photobleaching, it is possible to resolve the spectral bleed‐through and retrieve the FRET efficiency/interaction distance quantitatively. Using a human keratinocyte cell line transfected with cyan fluorescent protein (CFP)‐ and yellow fluorescent protein (YFP)‐tagged Cx26 connexins as an example, FRET information at homotypic gap junctions is measured and compared with well‐established methods. Results indicate that the new approach is sensitive, flexible, instrument independent and solely FRET dependent. It can achieve FRET estimations similar to that from a sensitized emission FRET method. This approach has a great advantage in providing the relative concentrations of the donor and the acceptor; this is, for example, very important in the comparative study of cell populations with variable expression levels.
Five‐colour in vivo imaging of neurons in Caenorhabditis elegans
Authors: HUTTER H. 
Published: July 2004

SummaryIn the last few years variants of the ‘green fluorescent protein’ (GFP) with different spectral properties have been generated. This has greatly increased the number of possible applications for these fluorochromes in cell biology. The significant overlap of the excitation and emission spectra of the different GFP variants imposes constraints on the number of variants that can be used simultaneously in a single sample. In particular, the two brightest variants, GFP and YFP, are difficult to separate spectrally. This study shows that GFP and YFP can be readily separated with little spectral overlap (cross‐talk) with the use of a confocal microscope equipped with an acusto‐optical beam splitter and freely adjustable emission windows. Under optimal recording conditions cross‐talk is less than 10%. Together with two other fluorescent proteins and the lipophilic dye DiD a total of five different colours can now be used simultaneously to label in vivo distinct anatomical structures such as neurons and their processes. Spatial resolution of the confocal microscope is sufficient to resolve the relative position of labelled axons within a single axon bundle. The use of five distinct marker dyes allows the in vivo analysis of the Caenorhabditis elegans nervous system at unprecedented resolution and richness in detail at the light microscopic level.
Silver deposition on freeze‐dried cells allows subcellular localization of cholesterol with imaging TOF‐SIMS
Authors: NYGREN H. , MALMBERG P. 
Published: July 2004

SummaryImaging time‐of‐flight secondary ion mass spectrometry (TOF‐SIMS) was used for characterization and subcellular localization of organic ions in leucocytes adhering to glass surfaces. The cells were fixed by freeze drying in 0.15 m ammonium formate buffer at pH 7.2–7.4. The freeze‐dried cells were sputter‐coated with silver, and the silver surface was analysed with imaging TOF‐SIMS. TOF‐SIMS spectra were recorded by scanning the primary ion beam over the analysis area and acquiring positive mass spectra of the ions leaving the surface. The relative brightness of each pixel within the analysis area reflects the signal intensity of a selected ion in that pixel. Data were collected separately at high mass resolution m/Δm > 7000 and at high lateral resolution (= 0.5 µm). The images were analysed by principal component analysis (PCA).The glass‐adhering cells showed a well defined attachment area with a diameter of up to 20 µm, and an equally well defined cell body, containing the nucleus, with a diameter of 8–10 µm. On the raw data images, the obtained cholesterol distributions were consistent with a higher cholesterol content of the cell membrane in the attachment area than in the cell body. Using PCA analysis, silver‐cationized molecular cholesterol was found localized mainly in the attachment area of the cells. Cholesterol was also seen at higher concentration in circular spots of ≤ 1 µm in diameter, probably representing caveolae.
Pre‐ and post‐thaw assessment of intracellular ice formation
Authors: ACKER J. P. , CROTEAU I. M. 
Published: July 2004

SummaryIntracellular ice formation (IIF) refers to the formation of ice crystals within cells during rapid freezing. To develop an understanding of the means by which intracellular ice forms and the mechanisms by which it damages cells and tissues requires techniques that combine real‐time assessment of ice nucleation and ice crystal growth with detailed assessments of cell structure and function. Intracellular ice formation has been detected in live samples using light scattering, freeze substitution and fluorescent detection. In this study we develop a method to correlate IIF with post‐thaw structural analyses by combining low temperature microscopy and freeze substitution. V79‐4 hamster fibroblasts were frozen on a low temperature microscope at various temperatures, IIF was visualized using the nucleic acid‐specific fluorophore SYTO 13™, then the samples were fixed (10% formaldehyde, 85% ethanol, 5% acetic acid) while still frozen. The monolayers were then thawed and stained with routine histological stains haematoxylin and eosin and assessed. Fixation allowed for the post‐thaw assessment of IIF and for subsequent histological processing to examine in detail the structural consequences of IIF. The post‐thaw identification of cells that form intracellular ice during freezing is a significant improvement to current methods used in low temperature biology.
Fine‐scale spatial patterns in populations of the clonal moss Hylocomium splendens partly reflect structuring processes in the boreal forest floor
Authors: Økland Rune Halvorsen , Bakkestuen Vegar 
Published: July 2004

Spatial patterns of important population or individual characteristics are expected to reflect structuring ecosystem processes to the extent that populations with strikingly different spatial patterns most likely have been structured by different processes. Few studies of spatial pattern in plant assemblages, bryophyte assemblages in particular, still exist and pattern has only vaguely been linked with process. In this study we describe fine‐scale spatial variation in size and the occurrence of terminated (non‐branching) segments in 21 Norwegian populations of the dominant boreal forest moss Hylocomium splendens, in search for general patterns of spatial structure.All Hylocomium splendens segments in the investigated plots were followed for a 10‐year period; their size (dry mass, estimated from in situ measurements) and fate (terminated or ramifying) were recorded annually. The spatial structure of size and terminating segments at scales from 0.5 to 16 cm was described for each population as fractal dimension profiles, derived from semi‐variograms. For about one half of the populations size and terminating segments could be assigned to a specific spatial pattern, the predicted outcome of one of three main structuring processes: (i) negative spatial dependence (fractal dimension F>3.0) on the scale of individuals (below 2 cm), indicative of negative interactions; (ii) positive spatial dependence (F<3.0) on the scale of individuals, indicative of positive interactions; and (iii) positive spatial dependence on broader (4–8 cm) scales, indicative of structuring by environmental factors or patchy disturbance. Patterns (i) and (iii) were observed both for size and terminated segments more often than expected. Fractal dimension profiles for size obtained separately for each year revealed temporal patterns of spatial structure that tended to be invariant over years. Negative spatial dependence of size, possibly due to large size difference between buried segments and other segments during self thinning, was typical of populations rapidly growing in number. Positive spatial dependence with a range of influence by the spatial process of 4–8 cm was observed in plots with sparse bryophyte cover and high cover of deciduous litter, probably due to accumulation of litter in depressions between shoots or groups of shoots. The main finding of this study is that different populations of the same species in the same type of ecosystem can be spatially structured in different ways. This accords with a model for the studied system as consisting of a mosaic of cells, of spatial extent of a few cm or larger, intergrading in space and temporally dynamic, the population characteristics of each cell being shaped by several structuring processes that vary, temporally and spatially, in their relative importance.
The impact of short‐term predator removal on vole dynamics in an arctic‐alpine landscape
Authors: Ekerholm Per , Oksanen Lauri , Oksanen Tarja , Schneider Michael 
Published: July 2004

During 1991–95, mammalian predators (weasel, Mustela nivalis, stoat, M. erminea, mink, M. vison, and red fox, Vulpes vulpes) were excluded in late summers from a 2 ha piece of a north Norwegian mountain slope. The exclosure extended from an outpost of luxuriant sub‐arctic birch forest to a typical arctic–alpine habitat complex, including productive willow scrublands, tundra heaths, dry ridges and snow‐beds. The exclosure thus encompassed the entire range of habitat conditions encountered in a typical north Fennoscandian mountain and tundra landscapes. During 1991–95, the exclosure was predator‐proof from late July to late September. In wintertime and in early summer, the exclosure was accessible to mammalian predators. Vole dynamics in the short‐term exclosure were compared to dynamics in five reference areas with similar habitat conditions.In 1991, when vole densities were rising in the area, neither collective vole densities nor densities of individual vole species differed significantly between the exclosure and the replicated controls. Spring densities of voles were never significantly different between the exclosure and the controls. With respect to autumnal densities of voles, however, the exclosure was a statistical outlier in the peak year 1992 and throughout the gradual decline phase of 1993–95. In the peak year, the difference in collective vole densities was modest (30%), but increased to two‐fold during the first two decline years and was almost four‐fold in the crash year of 1995. The strongest response was displayed by field voles (Microtus agrestis), hypothesized to be the pivotal prey species of weasels, especially by females and young individuals, i.e. by the functional categories especially sensitive to mammalian predation. These results are consistent with the hypothesis that predation plays a pivotal role for the regulation of herbivorous mammals in relatively productive arctic–alpine habitats.
Conspicuousness, not colour as foraging cue in plant–animal signalling
Authors: Schmidt Veronika , Martin Schaefer Hinrich , Winkler Hans 
Published: July 2004

The global prevalence of red and black fruits has still not been explained. Hypotheses based on innate consumer preferences have been tested and rejected. Though colour itself plays an important role in animal foraging, it is only one component of signals. Another major component are colour contrasts against background achieving the conspicuousness of signals. In order to evaluate which signal component determines consumers behaviour, we measured fruit colour and colour contrasts of 43 species against their natural background under ambient light conditions. Red and black fruits exhibit stronger contrasts and are therefore more conspicuous to consumers than fruits of other colours. Subsequently, trials were carried out to determine whether colour or conspicuousness influences avian food choice. Four bird species strongly preferred contrasting red–green or black–green over uni‐coloured red, green, or black fruit displays, while no preference for particular hues was found. We therefore hypothesize that conspicuousness determines avian food selection and define the contrast hypothesis: Diurnal dispersers select fruit colours based on their conspicuousness and not their colour itself.Because colour vision is an ancient trait, the entire heterogeneous group of frugivorous birds most likely perceives conspicuousness uniformly over evolutionary time spans. Conspicuousness has thus the potential to explain the global prevalence of red and black fruits.
Plant species identity and diversity effects on different trophic levels of nematodes in the soil food web
Authors: De Deyn Gerlinde B. , Raaijmakers Ciska E. , Van Ruijven Jasper , Berendse Frank , Van Der Putten Wim H. 
Published: July 2004

Previous studies on biodiversity and soil food web composition have mentioned plant species identity, as well as plant species diversity as the main factors affecting the abundance and diversity of soil organisms. However, most studies have been carried out under limitations of time, space, or appropriate controls. In order to further examine the relation between plant species diversity and the soil food web, we conducted a three‐year semi‐field experiment in which eight plant species (4 forb and 4 grass species) were grown in monocultures and mixtures of two, four and eight plant species. In addition there were communities with 16 plant species. We analyzed the abundance and identity of the nematodes in soil and roots, including feeding groups from various trophic levels (primary and secondary consumers, carnivores, and omnivores) in the soil food web.Plant species diversity and plant identity affected the diversity of nematodes. The effect of plant diversity was attributed to the complementarity in resource quality of the component plant species rather than to an increase in total resource quantity. The nematode diversity varied more between the different plant species than between different levels of plant species diversity, so that plant identity is more important than plant diversity. Nevertheless the nematode diversity in plant mixtures was higher than in any of the plant monocultures, due to the reduced dominance of the most abundant nematode taxa in the mixed plant communities. Plant species identity affected the abundances of the lower trophic consumer levels more than the higher trophic levels of nematodes. Plant species diversity and plant biomass did not affect nematode abundance. Our results, therefore, support the hypothesis that resource quality is more important than resource quantity for the diversity of soil food web components and that plant species identity is more important than plant diversity per se.
Can the assumption of a non‐random search improve our prediction of butterfly fluxes between resource patches?
Authors: Kindlmann Pavel , Aviron Stephanie , Burel Francoise , Ouin Annie 
Published: July 2004

Abstract.  1. Understanding dispersal patterns that enable small, spatially isolated populations to survive in fragmented landscapes has become an important issue in conservation biology and landscape management. However, for most of the species of interest it is not known whether dispersing individuals navigate or follow systematic search strategies, as opposed to moving randomly.2. Recently it was shown that individuals of the butterfly species Maniola jurtina do not seek resources by means of random flight. If true, this may be problematic for existing metapopulation models, including those based on the evolution of dispersal rates in metapopulations.3. The study tested to what extent the non‐random dispersal patterns described in the literature can explain M. jurtina fluxes in its natural habitat.4. A model based on literature assumptions of M. jurtina movements is presented in the work reported here, and its predictions are compared with 2 years of capture–recapture data on its fluxes in two landscapes.5. The model provides a good fit to the data and gives better predictions than the model based only on patch sizes and distances between patches.6. Thus, if data are available about the actual landscape under consideration, the model should be preferred to simpler approaches; however, in general theoretical considerations the simple approach based on patch size and the degree of its isolation will retain its value.
Foraging in nature: foraging efficiency and attentiveness in caterpillars with different diet breadths
Authors: Bernays E. A. , Singer M. S. , Rodrigues D. 
Published: July 2004

Abstract.  1. Seventy‐seven individual last‐instar caterpillars foraging in the field were examined for 6 h each. They represented four species of Arctiidae of similar size and habitat use. Two, Hypocrisias minima and Pygarctia roseicapitis, are specialists restricted to particular plant genera. The other two, Grammia geneura and Estigmene acrea, are extreme generalists that use many host plant species from multiple plant families.2. Parameters of behavioural efficiency were monitored. Generalists spent more time walking, rejected more potential host plants, took longer to decide to feed after inspecting a plant, and took relatively more small feeding bouts compared with specialists.3. This is the first test of differential foraging efficiency in the field in relation to diet breadth of insects and the data indicate that generalists are less efficient in their foraging activities and may suffer from divided attention. The need for attentiveness to enhance efficiency and thereby reduce ecological risk is discussed.
Effects of parental body condition and size on reproductive success in a tenebrionid beetle with biparental care
Authors: Heg Dik , Rasa O. Anne E. 
Published: July 2004

Abstract.  1. The beetle Parastizopus armaticeps (Coleoptera: Tenebrionidae) inhabits the Kalahari desert of southern Africa, constructs breeding burrows after rainfall, and shows extensive biparental care. Previous work has shown that it is predominantly male size, not female size, that determines breeding success; however, in the field these beetles show size assortative mating. This might obscure or override effects of female size on reproduction. Moreover, the inaccessibility of the breeding burrows makes it impossible to test effects of female and male size on offspring development and survival before adulthood.2. To disentangle the effects of male and female length, body mass, and body condition on reproductive success, males and females were paired randomly in small breeding cages in the laboratory (n = 887 breeding pairs). The construction of the breeding cages allowed a clear view of the brood chamber contents at each stage in offspring development. Larva, pupa, and imago numbers and development were monitored daily, and imago mass at hatching from the pupa (hatchlings), offspring mass, and offspring body length at complete exoskeleton melanisation (juveniles) were determined.3. There was a weak positive correlation between body condition and body length for females only. Breeding chronology was related to male body condition: males in better condition were fast to start and finish a breeding bout. Males in better condition produced heavier hatchlings and juveniles, and larger‐sized males produced larger‐sized juveniles. In contrast, numbers of larvae and juveniles produced were determined mainly by female length and body condition: larger females in better condition hatched more larvae and produced more offspring.4. The results suggest that male size and condition will be the most important determinant of reproductive success under relatively dry conditions, when burrow length is critical for reproductive success. Female size might be more important for the pair's reproductive success under wet breeding conditions, when burrow length is less critical for successful reproduction.