The Experts below are selected from a list of 14703 Experts worldwide ranked by ideXlab platform
Alfonso Barbarisi - One of the best experts on this subject based on the ideXlab platform.
-
Immunogold Labelling in environmental scanning electron microscopy applicative features for complementary cytological interpretation
Journal of Microscopy, 2011Co-Authors: Gennaro Cafiero, F Papale, Anna Grimaldi, Francesco Rosso, Manlio Barbarisi, Carla Tortora, Gerardo Marino, Alfonso BarbarisiAbstract:We have combined environmental scanning electron microscopy (ESEM) and Immunogold Labelling (IGL) for the analysis of cell morphology and surface protein detection on human fine needle aspiration, which is processed in thin uniform monolayer (a single layer of cells) on a glass slide by Thin Prep technology. Among scanning electron microscopy techniques, we choose the environmental modality (ESEM) because it allows a slight manipulation of biological samples and an operational time comparable with cytological techniques. Moreover, the Thin Prep technology confirmed a reproducible cell monolayer on glass smear, minimizing problems for the determination of appropriate amount of material per slide. The first experimental data in ESEM-IGL on biological samples with fine needle aspiration Thin Prep, in human thyroid nodules, showed that cells retained their morphology and provided a clear IGL. The optimization of conditions (i.e. vacuum pressure, temperature and relative humidity) confirmed the possibility to observe an immunolabelled biological sample and morphological signal, joined with compositional informations, due to peculiar characteristics of gaseous secondary electron detector in ESEM. The ESEM-IGL and fine needle aspiration Thin Prep could be used in combination for the interpretation of cell morphology and cell surface immunoLabelling. Our paper suggests this use as a powerful diagnostic tool in a pre-surgical evaluations, opening a new applicative window for electron microscopy.
-
cell surface protein detection with Immunogold Labelling in esem optimisation of the method and semi quantitative analysis
Journal of Cellular Physiology, 2008Co-Authors: Livio Muscariello, Gennaro Cafiero, Francesco Rosso, Manlio Barbarisi, Gerardo Marino, Alfonso BarbarisiAbstract:In this work we used a combination of Immunogold Labelling (IGL) and environmental scanning electron microscopy (ESEM) to detect the presence of a protein on the cell surface. To achieve this purpose we chose as experimental system 3T3 Swiss Albino Mouse Fibroblasts and galectin-3. This protein, whose sub-cellular distribution is still under discussion, is involved in a large number of cell physiological and pathological processes. IGL technique has been utilised by many authors in combination with SEM and TEM to obtain the identification/localisation of receptors and antigens, both in cells and tissues. ESEM represents an important tool in biomedical research, since it does not require any severe processing of the sample, lowering the risk of generating artefacts and interfere with IGL procedure. The absence of metal coating could yield further advantages for our purpose as the Labelling detection is based on the atomic number difference between Nanogold spheres and the biological material. Using the gaseous secondary electron detector (GSED) compositional contrast is easily revealed by the backscattered electrons component of the signal. In spite of this fact, only few published papers present a combination of ESEM and IGL. Hereby we present our method, optimised to improve the intensity and the specificity of the Labelling signal, in order to obtain a semi-quantitative evaluation of the Labelling signal. J. Cell. Physiol. 214: 769–776, 2008. © 2007 Wiley-Liss, Inc.
Elisabeth Grenet - One of the best experts on this subject based on the ideXlab platform.
-
Immunogold Labelling of feruloyl-arabinose of maize stem after rumen microbial degradation
Journal of the Science of Food and Agriculture, 1999Co-Authors: Carole Migné, Gérard Prensier, Agnès Cornu, J.p. Utille, P. Angibeaud, Elisabeth GrenetAbstract:Immunogold Labelling of feruloyl-arabinose was performed on the base and the top of the apical internode of the stem of Co125 and W401 maize, harvested at anthesis +5 days, after incubation in the rumen in nylon bags for 4, 8 and 24h. The description of Labelling evolution during rumen degradation was permitted by the quantification of the intensity of Labelling in four tissues: sclerenchyma, fibres, xylem and parenchyma. The results are discussed in relation to maize digestibility.
-
immunocytochemical localisation of para coumaric acid in the cell walls of maize stem after rumen microbial degradation
Journal of the Science of Food and Agriculture, 1998Co-Authors: Carole Migné, Gérard Prensier, Agnès Cornu, Elisabeth GrenetAbstract:Immunogold Labelling of para-coumaric acid was performed on the base and the top of the apical internode of the stem of Co125 and W401 maize, harvested at anthesis + 5 days, after incubation in the rumen in nylon bags for 4, 8 and 24 h. The description of Labelling evolution during rumen degradation was permitted by the quantification of the intensity of Labelling in four tissues: sclerenchyma, fibres (surrounding vascular bundles), xylem and parenchyma. The results are discussed in relation to maize digestibility.
-
Immunocytochemical localisation of para‐coumaric acid in the cell walls of maize stem after rumen microbial degradation
Journal of the Science of Food and Agriculture, 1998Co-Authors: Carole Migné, Gérard Prensier, Agnès Cornu, Elisabeth GrenetAbstract:Immunogold Labelling of para-coumaric acid was performed on the base and the top of the apical internode of the stem of Co125 and W401 maize, harvested at anthesis + 5 days, after incubation in the rumen in nylon bags for 4, 8 and 24 h. The description of Labelling evolution during rumen degradation was permitted by the quantification of the intensity of Labelling in four tissues: sclerenchyma, fibres (surrounding vascular bundles), xylem and parenchyma. The results are discussed in relation to maize digestibility.
-
Immunogold Labelling of arabinoxylans in the plant cell walls of normal and bm3 mutant maize
Biology of the Cell, 1991Co-Authors: P. Barry, Gérard Prensier, Elisabeth GrenetAbstract:Abstract Polyclonal antibodies directed against α, L -1.2-arabinofuranosyl poly-β, D -1.4-xylopyranosyl (degree of polymerization 130) have been raised from rabbits. The Immunogold Labelling in transmission electron microscopy (TEM) evidenced the arabinoxylans of the plant cell walls. Comparison between the stems of normal and mutant bm3 maize demonstrated a greater accessibility of arabinoxylans in the walls of the mutant maize. The method, specific and swift, allows us to specify the repartition in the different parts of the stem: sclerenchyma, fibers, parenchyma.
Kenneth Goldie - One of the best experts on this subject based on the ideXlab platform.
-
Amyloid fibril formation by human stefin B in vitro : Immunogold Labelling and comparison to stefin A
Biological chemistry, 2017Co-Authors: Vito Turk, Maruša Pompe Novak, Eva Žerovnik, Tina Zavašnik-bergant, Miha Škarabot, Nataša Kopitar-jerala, Igor Muševič, Maja Ravnikar, Kenneth GoldieAbstract:The mechanism by which proteins form amyloid fibrils is of high interest to the scientific community as its understanding could resolve questions relevant to conformational diseases. The structural and energetic basis of the process is still largely unknown. The main controversial issue is the co-existence of several protein conformations. Three models for the mechanism of protein fibrillogenesis have been proposed which need to be tested by experiments. In this report, amyloid fibrils grown from human stefin B (type 1 cystatin) are described. This physiologically relevant protein readily forms fibrils in vitro, in contrast to the homologue - human stefin A - which forms fibrils under extreme conditions only. In order to specifically label stefin B fibrils in vitro, rabbit polyclonal antibody and mouse monoclonal antibody A6/2 against human stefin B were used for Immunogold Labelling. Samples were examined by transmission electron microscopy. Fibrils of stefin B were strongly labelled using polyclonal antibody and Protein A gold, whereas no positive reaction was observed with monoclonal antibody A6/2.
Frédéric Marin - One of the best experts on this subject based on the ideXlab platform.
-
Chalky versus foliated: a discriminant Immunogold Labelling of shell microstructures in the edible oyster Crassostrea gigas
Marine Biology, 2016Co-Authors: Vincent Mouchi, Franck Lartaud, Nathalie Guichard, Françoise Immel, Marc De Rafélis, Cédric Broussard, Quentin G. Crowley, Frédéric MarinAbstract:Mollusc shells are organic–inorganic biocomposites, arranged in a limited number of superimposed calcified layers that generally exhibit very different organization of their crystallites. Because of their attractive mechanical and crystallographic properties, these shell layers have been the focus of several physical and biochemical characterizations. In particular, recent proteomic data obtained from individual layers suggest that their protein contents are different. However, the direct visual evidence that some macromolecular components are layer-specific is rather tenuous. This paper is based on a non-conventional Immunogold Labelling approach to localize proteins in the shell of the edible oyster Crassostrea gigas . The shell microstructure of this model organism is predominantly composed of foliated calcite, interspersed by discontinuous pockets of ‘chalky layers’, a porous microstructure typical of bivalves of the ostreid family. By developing a polyclonal antibody (in two rats) elicited against a proteinaceous shell fraction, we obtained differential staining of the two microstructures. We assert that our Labelling is microstructure discriminant. The difference in Labelling of the two shell microstructures suggests either that they are formed by a variation of the secretory repertoire of the shell-forming cells of the calcifying mantle epithelium or that the chalky layer may be formed via a completely different mechanism. Our results allow a first glimpse on the subtle regulatory mechanisms that drive the process of chalky and foliated layers deposition.
E Skarmoutsou - One of the best experts on this subject based on the ideXlab platform.
-
quantifying Immunogold Labelling in transmission electron microscopy
Journal of Microscopy, 2008Co-Authors: F Damico, E SkarmoutsouAbstract:Summary Gold particles Labelling on ultrathin sections is extensively used for antigen localization in transmission electron microscopy. In establishing absolute or relative counts in tissue sections, it would be expedient to use stereologically based unbiased estimates for quantitative results. Nowadays, quantitative immunoelectron microscopy has achieved good and satisfactory results to test whether the gold Labelling follows a non-random or a random pattern and then to draw statistical comparisons between cell subcompartments within a sample of cells or between experimental groups of cells. This brief informal review of literature focuses on the relative quantitative determinations of gold Labelling of antigens as well as on the statistical distribution comparisons in transmission electron microscopy.
