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Pergamon Press

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    Can insecticide-free clean water regenerate the midgut epithelium of the freshwater shrimp after dimethoate treatment?

    Ostrozka, A.Tiffert, Z.Wilczek, G.Rost-Roszkowska, M....
    16页
    查看更多>>摘要:Insecticides such as dimethoate persist for a long time in freshwater environments, influencing the physiology of the animals inhabiting such environments. In aquatic organisms, toxic substances can enter the body through the epidermis and the digestive system. The midgut is part of this system in which intense processes constitute a barrier against the effects of toxic substances on the body. The aim of this study was to evaluate the toxic potential of dimethoate in the midgut epithelium of the freshwater shrimp Neocaridina davidi, emphasizing ultra structural alterations. However, the additional and main purpose was to determine whether the midgut epithelium can regenerate after placing animals in insecticide-free clean water after various periods of exposure to dimethoate. N. davidi originates from Asia, but it has also been described in European rivers. This species is of particular interest among breeders worldwide due to its ease of breeding and reproduction. The animals were treated with dimethoate for 1, 2, and 3 weeks and then placed in clean water for 1, 2, and 3 weeks. The qualitative and quantitative analysis revealed different sensitivity of organs forming the midgut in freshwater crustaceans and the possibility for midgut regeneration after insecticide exposure. We concluded that different processes were triggered in the intestine and hepatopancreas to regenerate cells after damage, and mitochondria were the first organelles to respond to the appearance of a stressor in the living environment.

    Atomic force spectroscopy is a promising tool to study contractile properties of cardiac cells

    Kabanov, DaniilKlimovic, SimonRotrekl, VladimirPesl, Martin...
    8页
    查看更多>>摘要:Atomic Force Microscopy (AFM) is a rather new method with increasing potential in analyzing various biosamples. Moreover, it can serve as a multi-functional device in the studies of biological specimens under physiological conditions. However, it is becoming increasingly popular among biochemists and biologists, it is not often used in cardiology. Heart disease causes millions of deaths every year. A common point in all heart diseases is the inferior function of cardiomyocytes, which are the contracting unit of the heart. Therefore, these cells are a frequent target of scientific studies. However, few of them use innovative techniques such as AFM and related methods or parallel combinations with complementary techniques such as cell potential measurements. The aim of this review is to illustrate the potential of AFM microscopy in the study of cardiac cells, comparing it with related methods and other techniques used to study the biomechanics and electrophysiology of this cell type. A better understanding of these methods may lead to a better description of the pathophysiology of the heart disease and an improved understanding of the effect of selected drugs.

    Development of a direct point electron beam exposure system to investigate the biological functions of subcellular domains in a living biological cell

    Tanaka, AsahiInami, WataruSuzuki, YukoKawata, Yoshimasa...
    7页
    查看更多>>摘要:Electron microscopy studies have demonstrated that the diameter of a focused electron beam is small enough to probe or manipulate subcellular domains of a single biological cell. Here, we report the development of a direct point electron beam irradiation system to investigate the biological functions of subcellular domains in a living cell. Subcellular structures of a single living cell cultured on a thin film can be selectively irradiated by the point electron beam generated by our system. We have demonstrated controlled beam positioning capability to selectively irradiate 500 nm size structure with a point electron beam. We determined beam irradiation parameters that did not cause irreversible plasma membrane perforation after beam exposure and the irradiation caused intracellular Ca2+ elevation in an irradiated neuronal cell. Since the neuronal cell express fine subcellular structures such as neurites, we tried to position a beam on the structure and observed a Ca2+ wave originated from the intended point, which showed that our system had enough selectivity to target a subcellular structure. Point electron beam exposure is expected to be employed for various cellular stimulation protocols, and this enables the investigation of the biological functions of subcellular domains.

    Transmission electron microscopy of precipitation in fine-grained heat-affected zone of Grade91 steel weld during creep exposure

    Peansukmanee, SompornPhung-on, IsaratatPoopat, BovornchokTsuda, Kenji...
    15页
    查看更多>>摘要:Precipitation during creep exposure at 550 degrees C in the fine grain heat-affected zone of a Grade91 steel weld produced by gas tungsten arc welding with no filler and post-weld heat treatment at 760 degrees C for 30 min. has been investigated. The applied tension load during creep testing was 165 MPa, the creep rupture time was 1138 hrs. (100% creep aging), and the creep exposure times were 911, 569, 341, 223 and 114 hrs. for 80%, 50%, 30%, 20% and 10% creep aging, respectively. Light microscopy and scanning electron microscopy were used to examine the heat-affected zone in the steel weld. Grain size in the fine grain heat-affected zone was measured by a circular intercept method on secondary electron images from scanning electron microscopy. Precipitates were detached from the fine grain heat-affected zone by a carbon-film extraction replica technique and investigated by transmission electron microscopy. During creep, a continual increase in grain size was observed in the fine grain heat-affected zone, starting from about 1.70 mu m after post-weld heat treatment to about 2.30 mu m at 100% creep aging. Precipitates found during creep were M23C6, M7C3, M6C, MX, M2X and high-Si Laves phase. M23C6 was found in all creep conditions until rupture, mainly at prior austenite grain boundaries and sub-boundaries. M7C3, M6C and high-Si Laves phase were observed during creep aging, mainly nearby grain boundaries and sub boundaries. Relatively fine MX and M2X were found after 20% creep aging until rupture, mainly within grain interiors. Detailed criteria for identification of these precipitate phases, based mainly on energy dispersive X-ray spectrometry and selected area electron diffraction using transmission electron microscopy, is given and a major cause of premature loss of creep strength in Grade91 steel weld is discussed. This is important in understanding creep behavior of steel parts used for boiler tubing, heat exchangers, and steam piping systems in fossil-fuel power plants.

    A method for cleaning flat punch diamond microprobe tips

    Everaerts, JorisSlagter, AlejandraMortensen, Andreas
    4页
    查看更多>>摘要:Microprobe tips are commonly used to perform in-situ micromechanical tests within an electron microscope. In service, such tips have a tendency to accumulate along their surface a layer of deposited material. Tip cleanliness is crucial in order to obtain reliable and reproducible data; however, cleaning of such tips can be arduous, due to their fragility. The literature on appropriate tip cleaning methods is relatively sparse; we aim in this study to fill this gap by presenting an effective way to clean flat punch diamond microprobe tips within an electron microscope, based on mechanical scraping. Initial attempts to remove deposits from a contaminated diamond tip using two micro-brush samples, one containing silica needles and the other containing cementite lamellae, were unsuccessful, due to the adherence of the deposit to the surface of the tip and its apparently high hardness. The successful cleaning method consists of milling a silicon ridge by means of a focused ion beam, and then using this ridge to effectively scrape the deposits off the tip surface in a controlled and complete manner. This method avoids potential damage to the microprobe and can be implemented easily to clean flat punch tips rapidly within a scanning electron microscope.

    Ultrastructural studies of photoreceptor cell degeneration with organophosphate and its regeneration in Cyprinus carpio communis L.

    Kaur, RavneetGarg, VincyJohal, M. S.
    10页
    查看更多>>摘要:Organophosphates are highly neurotoxic to aquatic fauna if they enter the water bodies as runoff, thus affecting the nervous system of the fishes. The present study was undertaken to investigate the vision changes, especially on the photoreceptor layer of the retina, of Cyprinus carpio communis L. when exposed to monocrotophos, an organophosphate. Fish were exposed to three sub-lethal concentrations of LC50, i.e. 0.038 ppm (1/10 LC50), 0.062 ppm (1/6 LC50), and 0.126 ppm (1/3 LC50), to observe the changes in the photoreceptor cells at the behavioral, histopathological and ultrastructural levels. Further, acetylcholinesterase activity was also evaluated. Behavioral changes, such as long resting period, inactivity, increase in air gulps and decrease in opercular and fin movements, were observed. A semi-quantitative analysis of the histological sections showed shrinkage in retinal layers at 0.038 ppm concentration of monocrotophos. At 0.062 ppm, the disappearance of the outer nuclear layer was observed and at the highest concentration of 0.126 ppm, damage in all retinal layers involving necrosis of the outer segment of the photoreceptor cells was observed. Further, at the ultrastructural level, detachment of photoreceptor cells and damage in the inner and outer segments of the photoreceptors were observed in an increasing dose-dependent manner. A reduction in the acetylcholinesterase level was observed in the treated groups. The treated fish were then transferred to toxicant-free water for 60 days to study self-regeneration, but no regeneration was observed in photoreceptor cells of the fish retina. This study shows that exposure to of monocrotophos effectively damages and disturbs the functioning of photoreceptor cells of retina of C. carpio communis L., thus affecting its vision.

    The Hertzian theory in AFM nanoindentation experiments regarding biological samples: Overcoming limitations in data processing

    Kontomaris, S. V.Malamou, A.Stylianou, A.
    13页
    查看更多>>摘要:Atomic Force Microscopy (AFM) nanoindentation is a powerful tool for the mechanical nano-characterization of biological samples. However, the range of Young's modulus values for the same type of samples usually varies significantly in the literature. This fact is partly related to the inhomogeneity of biological samples at the nanoscale and partly to significant mistakes during data processing. This review depicts that common errors related to (i) the real shape of the AFM tip, (ii) the range of data for which the sample presents an approximate linear elastic response, (iii) the sample's viscoelasticity, (iv) the sample's shape and (v) the substrate effects can be easily avoided without increasing the complexity of data processing. Thus, the present review paper focuses on the procedures that should be followed for the accurate processing of force-indentation curves regarding experiments on biological samples.

    Effect of antibiotics on mechanical properties of Bordetella pertussis examined by atomic force microscopy

    Villalba, M. I.Venturelli, L.Arnal, L.Masson, C....
    9页
    查看更多>>摘要:In recent years, the coevolution of microorganisms with current antibiotics has increased the mechanisms of bacterial resistance, generating a major health problem worldwide. Bordetella pertussis is a bacterium that causes whooping cough and is capable of adopting different states of virulence, i.e. virulent or avirulent states. In this study, we explored the nanomechanical properties of both virulent and avirulent B. pertussis as exposed to various antibiotics. The nanomechanical studies highlighted that only virulent B. pertussis cells undergo a decrease in their cell elastic modulus and height upon antimicrobial exposure, whereas their avirulent counterparts remain unaffected. This study also permitted to highlight different mechanical properties of individual cells as compared to those growing in close contact with other individuals. In addition, we analyzed the presence on the bacterial cell wall of Filamentous hemagglutinin adhesin (FHA), the major attachment factor produced by virulent Bordetella spp., under different virulence conditions by Force Spectroscopy.

    Fast determination of sample thickness through scanning moire fringes in scanning transmission electron microscopy

    Nan, PengfeiLiang, ZhiyaoZhang, YueLiu, Yangrui...
    5页
    查看更多>>摘要:Sample thickness is an important parameter in transmission electron microscopy (TEM) imaging for interpreting image contrast and understanding the relationship between properties and microstructure. In this study, we introduce a method for sample thickness determination in scanning TEM (STEM) mode based on scanning moire fringes (SMFs). Focal-series SMF imaging is used and sample thickness can be determined in situ at a medium magnification range, with beam damage and contamination avoided to a large extent. It provides a fast and convenient approach for determining sample thickness in TEM imaging, which is particularly useful for beamsensitive materials.