Microscopy Research and Technique via MedWorm.com MedWorm.com provides a medical RSS filtering service. Over 6000 RSS medical sources are combined and output via different filters. This feed contains the latest items from the 'Microscopy Research and Technique' source. Sun, 21 Mar 2010 17:30:16 +0100 Calibration issues for nanoindentation experiments: Direct atomic force microscopy measurements and indirect methods http://www.medworm.com/index.php?rid=3385251&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20850 This article discusses calibration issues for shallow depth nanoindentation experiments with Berkovich tips with respect to the accurate measurement of the diamond area function (DAF). For this purpose, two different calibration procedures are compared: (i) the direct measurement of the DAF through atomic force microscopy (AFM) imaging of the Berkovich tip at shallow depth and (ii) a novel indirect calibration method based on an iterative robust and converging scheme in which both reduced modulus and indentation hardness are simultaneously used. These results are obtained by indentation measurements on a standard specimen of fused silica, performed in the 0.5-200 mN load range with a Berkovich indenter. Direct tip shape measurements were carried out through different AFM methods. Compariso... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3385251 Sat, 20 Mar 2010 00:00:00 +0100 3385251 Bioluminescent imaging of reporter gene expression in the lungs of wildtype and model mice following the administration of PEG-stabilized DNA nanoparticles http://www.medworm.com/index.php?rid=3385252&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20855 DNA nanoparticles (DNPs) formed by compacting DNA with polyethyleneglycolylated poly-L-lysine are a nonviral vector shown to be safe and efficacious in animals and humans. To extend our capabilities of assessing the efficacy and duration of expression achieved by DNPs, we tested the utility of bioluminescent imaging (BLI) of transgene expression in wildtype and cystic fibrosis (CF) mouse models. We tested the effect of route of administration, mouse coat color, anesthesia, dose, and promoter sequence on the level and duration of expression. Furthermore, we investigated the correlation between imaging and direct analysis of luciferase expression in lung homogenates. We found that intratracheal instillation, and the use of deep and prolonged anesthesia with avertin produced significantly hig... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3385252 Fri, 19 Mar 2010 00:00:00 +0100 3385252 Immunochemical or fluorescent labeling of vesicular subcellular fractions for microscopy imaging http://www.medworm.com/index.php?rid=3370215&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20852 We describe a procedure for the labeling of membranous vesicular purified subcellular fractions, to image them, typically by confocal laser scanning microscopy. Being intracellular organelles, these fractions, once purified cannot be attached to glass slides as for cells. Fractions are labeled "in batch" without prior embedding or freezing. Each labeling step performed by passages of resuspension/centrifugation is followed by washings. Then samples are dispersed on the glass slides. Mammalian retinal rod outer segment disks, intact brain stem myelin vesicles, and brain synaptosomes were chosen, as these subcellular fractions can be purified by well established procedures. These fractions were immunolabeled with specific antibodies. Moreover, by the earlier procedure, we show that the mitoc... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3370215 Tue, 16 Mar 2010 00:00:00 +0100 3370215 Imaging multiple intermediates of single-virus membrane fusion mediated by distinct fusion proteins http://www.medworm.com/index.php?rid=3370231&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20841 Membrane fusion plays an essential role in the entry of enveloped viruses into target cells. The merging of viral and target cell membranes is catalyzed by viral fusion proteins, which involves multiple sequential steps in the fusion process. However, the fusion mechanisms mediated by different fusion proteins involve multiple transient intermediates that have not been well characterized. Here, we report a synthetic virus platform that allows us to better understand the different fusion mechanisms driven by the diverse types fusion proteins. The platform consists of lentiviral particles coenveloped with a surface antibody, which serves as the binding protein, along with a fusion protein derived from either influenza virus (HAmu) or Sindbis virus (SINmu). By using a single virus tracking te... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3370231 Mon, 15 Mar 2010 00:00:00 +0100 3370231 Unbiased stereological estimation of the rat fetal pituitary volume and of the total number and volume of TSH cells after maternal dexamethasone application http://www.medworm.com/index.php?rid=3370230&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20842 Glucocorticoids have an inhibitory influence on proliferation activity of the pituitary cells while stimulating apoptosis. Therefore, it was hypothesized that the synthetic glucocorticoid, dexamethasone (DX), has an inhibitory influence on the number of thyroid-stimulating hormone (TSH) cells during fetal development. The effects of maternal administration of DX on stereological parameters of TSH cells, and TSH serum concentration were investigated in 21-day-old rat fetuses. On day 16 of pregnancy, the experimental dams received 1.0 mg DX/kg b.w. subcutaneously, followed by 0.5 mg DX/kg b.w./day on days 17 and 18 of gestation. The control gravid females received the same volume of saline vehicle. TSH cells were stained immunocytochemically by the peroxidase-antiperoxidase (PAP) method. The... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3370230 Mon, 15 Mar 2010 00:00:00 +0100 3370230 Electron microscopy studies of electron-beam sensitive PbTe-based nanostructures http://www.medworm.com/index.php?rid=3370229&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20843 PbTe nanocrystals were synthesized and then reacted with a toluene solution containing AuCl3. Depending on the reaction conditions and on the starting PbTe nanocrystals morphology, different kinds of nanostructures have been obtained: single defect-free PbTe nanocrystals with and without an external amourphous oxide shell, crystalline-Au core/amorphous PbxTeyAuz shell structure, large balloon-shaped (or mushroom-shaped) Au domain attached via its apex to the surface of the nanocrystals. Structure and composition of these different types of nanostructure have been studied by means of Cs-corrected High Resolution Transmission Electron Microscopy (HRTEM) and Scanning Transmission Electron Microscopy (STEM) together with Energy Dispersive X-Ray Spectrometry (EDX), respectively. These nanostruc... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3370229 Mon, 15 Mar 2010 00:00:00 +0100 3370229 Effect of encapsulation or grafting on release kinetics of recombinant human bone morphogenetic protein-2 from self-assembled poly(lactide-co-glycolide ethylene oxide fumarate) nanoparticles http://www.medworm.com/index.php?rid=3370228&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20846 The objective of this work was to compare the release characteristics of Recombinant human bone morphogenetic protein-2 (rhBMP-2) encapsulated in thermally self-assembled poly(lactide ethylene oxide fumarate) (PLEOF) nanoparticles (NPs) with rhBMP-2 grafted to succinimide-terminated poly(lactide fumarate) (PLAF-NHS) or poly(lactide-co-glycolide fumarate) (PLGF-NHS) NPs. The amphiphilic PLEOF NPs had average size of 110 ± 50 nm. The hydrophobic PLAF-NHS and PLGF-NHS NPs had average size of 242 ± 67 and 195 ± 42 nm, respectively. PLEOF NPs had rhBMP-2 encapsulation efficiency ranging from 65 to 93%. Grafting efficiency of rhBMP-2 to PLAF-NHS and PLGF-NHS NPs was 97% ± 1% and 98% ± 1%, respectively. PLEOF NPs displayed a relatively high-release rate of rhBMP-2 in the first week, which ra... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3370228 Mon, 15 Mar 2010 00:00:00 +0100 3370228 Multiscale observation of biological interactions of nanocarriers: From nano to macro http://www.medworm.com/index.php?rid=3370227&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20847 Microscopic observations have played a key role in recent advancements in nanotechnology-based biomedical sciences. In particular, multiscale observation is necessary to fully understand the nano-bio interfaces where a large amount of unprecedented phenomena have been reported. This review describes how to address the physicochemical and biological interactions of nanocarriers within the biological environments using microscopic tools. The imaging techniques are categorized based on the size scale of detection. For observation of the nanoscale biological interactions of nanocarriers, we discuss atomic force microscopy (AFM), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). For the micro to macro-scale (in vitro and in vivo) observation, we focus on confocal l... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3370227 Mon, 15 Mar 2010 00:00:00 +0100 3370227 Combination of atomic force microscopy and principal component analysis as a general method for direct recognition of functional and structural domains in nanonocomposite materials http://www.medworm.com/index.php?rid=3370226&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20837 In this work, we report a simple method to direct identify nanometer sized textures in composite materials by means of AFM spectroscopy, aiming at recognizing structured region to be further investigated. It consists in acquiring a set of dynamic data organized in spectroscopy maps and subsequently extracting most valuable information by means of the principal component analysis (PCA) method. This algorithm projects the information of D spectroscopy curves, each containing P data, acquired at each point of an LxC grid into a subset of LxC maps without any assumption on the sample structure, filtering out redundancies and noise. As a consequence, a huge amount of 3D data is condensed into few 2D maps, easy to be examined. Results of this algorithm allow to find and locate regions of interes... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3370226 Mon, 15 Mar 2010 00:00:00 +0100 3370226 AFM measurement of the stiffness of layers of agarose gel patterned with polylysine http://www.medworm.com/index.php?rid=3370225&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20838 Films of agarose gel microspotted with polylysine aqueous solution have been characterized by atomic force microscopy carried out in deionized water. Thickness and surface morphology of the layers have been checked, and the effect of polylysine impregnation on the local elasticity has been investigated. An increase in contact stiffness of the organic layer at the spotted areas has been observed, correlated with the polylysine concentration. For the considered agarose layer thickness of [sim]0.9 [mu]m in dry condition, the concentration threshold at which stiffening appears is [sim]0.1 mg/mL. Above this threshold, the stiffening coefficient becomes approximately twofold and seems not to increase significantly with concentration in the range 0.3-0.7 mg/mL. For concentrations above the stiffe... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3370225 Mon, 15 Mar 2010 00:00:00 +0100 3370225 3D reconstruction of histological sections: Application to mammary gland tissue http://www.medworm.com/index.php?rid=3370224&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20829 We present an experimental validation of this tool using both synthetic and real data. In particular, we present the 3D reconstruction of an entire mouse mammary gland and demonstrate the integration of high-resolution molecular data. Microsc. Res. Tech., 2010. © 2010 Wiley-Liss, Inc. (Source: Microscopy Research and Technique) Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3370224 Mon, 15 Mar 2010 00:00:00 +0100 3370224 Structural and functional consequences of poly(ethylene glycol) inclusion on DNA condensation for gene delivery http://www.medworm.com/index.php?rid=3370223&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20839 Polycationic polymers have been used to condense therapeutic DNA into submicron particles, offering protection from shear-induced or enzymatic degradation. However, the spontaneous nature of this self-assembly process gives rise to the formation of multimolecular aggregates, resulting in significant polyplex heterogeneity. Additionally, cytotoxicity issues and serum instability have limited the in vivo efficacy of such systems. One way these issues can be addressed is through the inclusion of poly(ethylene glycol) (PEG). PEG has known steric effects that inhibit polyplex self-aggregation. A variety of PEGylated gene delivery formulations have been previously pursued in an effort to take advantage of this material's benefits. Because of such interest, our aim was to further explore the cons... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3370223 Mon, 15 Mar 2010 00:00:00 +0100 3370223 Cellular uptake pathway and drug release characteristics of drug-encapsulated glycol chitosan nanoparticles in live cells http://www.medworm.com/index.php?rid=3370222&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20845 Herein, we evaluated the cellular uptake pathways of hydrophobically modified glycol chitosan (HGC) nanoparticles as nano-sized drug carriers using cellular imaging technology. The endocytic pathway of nanocarriers for intracellular drug delivery is of great interest for the design of high efficacy delivery carriers for therapeutic agents. To evaluate the cellular uptake pathways of HGC nanoparticles, HGC was chemically labeled with near infrared (NIR) fluorescence dye, Cy5.5, to visualize the nanoparticle under confocal laser scanning microscopy. The internalization pathways of HGC nanoparticles were evaluated after treatment of specific endocytosis inhibitors. Importantly, HCG nanoparticles showed different cellular uptake efficiency and intracellular fate in cytoplasm according to the i... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3370222 Mon, 15 Mar 2010 00:00:00 +0100 3370222 Formation and microscopic investigation of iron oxide aligned nanowires into polymeric nanocomposite films http://www.medworm.com/index.php?rid=3370221&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20848 We present a microscopic investigation of nanocomposite films of iron oxide ([gamma]-Fe2O3) magnetic nanowires (NWs) aligned into polymers, formed upon evaporation of solutions of acrylate polymer/magnetic nanoparticles under magnetic field (MF). The field causes the assembly of the [gamma]-Fe2O3 nanoparticles along the direction of the MF lines, resulting in magnetic NWs embedded throughout the entire volume of the polymer film. The scanning electron microscopy and the transmission electron microscopy studies show that the cylindrical-shaped NWs have [sim]15-[mu]m average length and are isotropically distributed throughout the film. The study with the MF microscopy technique not only proves that the composed NWs are magnetic but also makes possible the magnetic study of each individual NW... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3370221 Mon, 15 Mar 2010 00:00:00 +0100 3370221 Force spectroscopy as a tool to investigate the properties of supported lipid membranes http://www.medworm.com/index.php?rid=3370220&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20834 Solid supported lipid bilayers (SLB) are extensively used as a model for the investigation of cell membranes in a variety of spectroscopic and biophysical methods. It is nevertheless well known that the interaction with the solid substrate, such as mica or silicon, influences the properties of the membranes. In this article we have employed atomic force microscopy (AFM) in force spectroscopy mode (FS) to investigate the local mechanical properties of lipid membranes supported on mica and on polymer cushion. The lipid double layers were obtained by fusion of unilamellar vesicle of phospholipids. The polymer support was created by self-assembly of charged polyelectrolytes. Force spectroscopy provided information about the breakthrough force, the breakthrough depth, and the sample adhesion. A... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3370220 Mon, 15 Mar 2010 00:00:00 +0100 3370220 Two and three-dimensional gene transfer from enzymatically degradable hydrogel scaffolds http://www.medworm.com/index.php?rid=3370219&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20840 In this report, we explored gene transfer to MSCs seeded on top or inside matrix metalloproteinase (MMP) degradable hydrogels that were loaded with DNA/poly(ethylene imine) (PEI) polyplexes. DNA/PEI polyplexes were encapsulated inside poly(ethylene glycol) (PEG) hydrogels crosslinked with MMP degradable peptides via Michael Addition chemistry. Gene transfer was visualized and quantified through using a vector encoding for green fluorescent protein and luciferase. We found that gene transfer to MSCs was possible for cells seeded both in two and three dimensions. The amount of luciferase expression was similar for cells seeded in two and three dimensions even though the number of cells in three dimensions is significantly higher, indicating that gene transfer to cells seeded in two dimension... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3370219 Mon, 15 Mar 2010 00:00:00 +0100 3370219 Rationalized approach to the determination of contact point in force-distance curves: Application to polymer brushes in salt solutions and in water http://www.medworm.com/index.php?rid=3370218&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20851 In this work, we present two methods to determine the contact point in force-distance curves obtained with the atomic force microscope. These procedures are compared with the typical determination of contact point by a visual assessment of the data. One method, based on the assumption that the sample shows linear elastic behavior, provides results similar to those obtained by a visual assessment of the data, and will be suitable for determining the contact point in cases where ionic repulsion is not significant. The second method is based on a series of measurements in which the sample deformation is measured at increasing values of applied load; the contact point is determined by extrapolation to zero load. Because this method is based on extrapolation of measurements made in the contact ... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3370218 Mon, 15 Mar 2010 00:00:00 +0100 3370218 Time sequence of blood activation by nanoporous alumina: Studies on platelets and complement system http://www.medworm.com/index.php?rid=3370217&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20854 In the present work, the time sequence of blood activation by alumina membranes with different porosities (20 and 200 nm in diameter) was studied. The membranes were incubated with whole blood from 2 min to 4 h. Platelet adhesion and activation in addition to complement activation was monitored at different time points. Evaluation of platelet adhesion and activation was done by determining the change in platelet number and the levels of thrombospondin-1 (TSP-1) in the fluid phase. Scanning electron microscopy studies were done to further evaluate platelet adhesion and morphology. Immunocytochemical staining was used to evaluate the presence of CD41 and CD62P antigens on the material surface. Complement activation was monitored by measuring C3a and sC5b-9 in plasma samples by means of enzym... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3370217 Mon, 15 Mar 2010 00:00:00 +0100 3370217 FT-IR, Raman, RRS measurements and DFT calculation for doxorubicin http://www.medworm.com/index.php?rid=3370216&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20849 Doxorubicin (DOXO) is a powerful anthracycline antibiotic used to treat many human neoplasms, including acute leukemias, lymphomas, stomach, breast and ovarian cancer, and bone tumors, yet causing cardiotoxicity at the same time. For this reason, there is a great interest in medical field to gain deep insight and knowledge of this molecule. Raman, Fourier Transform Infrared (FT-IR) absorption spectroscopy, and Resonance Raman scattering were performed for the vibrational characterization of DOXO molecule. Density function theorem (DFT) modeling of Raman and FT-IR spectra were used for the assignment of the vibrational frequencies. The optimized molecular structured was obtained, first, on the basis of potential energy distribution. The simulation for vibrational bands is based on the calcu... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3370216 Mon, 15 Mar 2010 00:00:00 +0100 3370216 Optimization of culture conditions for an efficient xeno-feeder free limbal cell culture system towards ocular surface regeneration http://www.medworm.com/index.php?rid=3318170&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20831 This study was aimed to establish an efficient xeno-feeder free limbal culture system towards ocular surface regeneration. To study the effect of initial dispase treatment and culture system used, migratory distance of cells from explants was analyzed from phase contrast images using "interactive measurements" of Qwin software (Leica). Expression of p63 in different culture systems was studied by immunofluorescent staining, followed by quantitative confocal microscopy (Carl Zeiss). Results showed dispase treatment was not necessary for establishing limbal explant culture. A combination of Iscove's modified Dulbecco's medium and Panserin 801 resulted in formation of autofeeder layer with maintenance of progenitor characteristics, thus mimicking natural tissue architecture. Further analysis ... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3318170 Mon, 01 Mar 2010 00:00:00 +0100 3318170 A scanning electron microscopy specimen holder for viewing different angles of a single specimen http://www.medworm.com/index.php?rid=3318169&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20835 The specimen holder for scanning electron microscopy described herein allows a single specimen to be examined in any possible view and significantly improves object illumination. The specimen is glued to a fine pin and flexibly mounted on a double-sided adhesive conductive pad on a rotatable pivot. A milled pot placed beneath the specimen acts as an electron trap. This provides a homogeneous black image background by minimizing noisy signals from the specimen's surroundings. Microsc. Res. Tech., 2010. © 2010 Wiley-Liss, Inc. (Source: Microscopy Research and Technique) Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3318169 Mon, 01 Mar 2010 00:00:00 +0100 3318169 Low-temperature glycol methacrylate resin embedding method: A protocol suitable for bone marrow immunohistochemistry, PCR, and FISH analysis http://www.medworm.com/index.php?rid=3318168&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20836 Molecular analyses such as fluorescence in situ hybridization (FISH) and polymerase chain reaction (PCR) are demanded to improve diagnostic accuracy in addition to immunohistopathology of bone marrow (BM) trephine specimens. Conventional BM embedding method needs decalcification, and its procedure may impair tissue morphology and DNA quality. Here, we report an undecalcified method by which glycol methacrylate resin is polymerized at low temperature (4°C). Using this method, BM enzyme activity and antigenic determinants are well preserved, and moreover, DNA extracted from plastic embedding sections is suitable for PCR amplification and sequencing, FISH analysis can be well done because of the DNA integrity of BM sections. If working with BM trephine specimen, our protocol offers the possi... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3318168 Mon, 01 Mar 2010 00:00:00 +0100 3318168 Effect of indirubin-3-monoxime against lung cancer as evaluated by histological and transmission electron microscopic studies http://www.medworm.com/index.php?rid=3287473&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20832 The aim of this study is to evaluate the antitumor effect of indirubin-3-monoxime and its mode of action in benzo([alpha])pyrene [B([alpha])P] induced lung cancer in A/J mice. Light microscopic examination of lung sections of [B([alpha])P] induced lung cancer mice revealed the presence of adenocarcinoma characterized by extensive proliferation of alveolar epithelium and loss of alveolar spaces. The control lung tissue showed a normal architecture with clear alveolar spaces. Interestingly the indirubin-3-monoxime treated groups showed the reduced adenocarcinoma with appearance of alveolar spaces. Transmission Electron Microscopic (TEM) studies of lung sections of [B([alpha])P] induced lung cancer mice showed the presence of phaemorphic cells with dense granules and increased mitochondria. T... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3287473 Fri, 19 Feb 2010 00:00:00 +0100 3287473 Early history, discovery, and expression of Aequorea green fluorescent protein, with a note on an unfinished experiment http://www.medworm.com/index.php?rid=3287476&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20821 This article reviews the work that took place leading to the discovery, cloning, and expression of the green fluorescent protein, with a note on an unfinished experiment. Microsc. Res. Tech., 2010. © 2010 Wiley-Liss, Inc. (Source: Microscopy Research and Technique) Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3287476 Thu, 18 Feb 2010 00:00:00 +0100 3287476 Two different pathways for the transport of primitive and definitive blood cells from the yolk sac to the embryo in humans http://www.medworm.com/index.php?rid=3287475&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20823 During the early human embryonic period nutrients and blood cells are temporarily provided by the extraembryonic yolk sac (YS). The YS before week six is involved not only in primitive but also in definitive erythropoiesis. While the destiny of primitive erythroid cells that fill the blood vessels of the YS is well known, the final destination of erythrocytes present in the endodermal vesicular system is unknown. In the present study we have investigated, step by step, the destiny of the erythrocytes present in the endodermal vesicles during the embryonic period.Twelve human YSs and their corresponding yolk stalks were analyzed between weeks 4 and 7 of embryonic age by light and scanning electron microscopy. It is shown that erythrocytes (according to their size and morphological features)... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3287475 Thu, 18 Feb 2010 00:00:00 +0100 3287475 Histological analysis of the alterations on cortical bone channels network after radiotherapy: A rabbit study http://www.medworm.com/index.php?rid=3287474&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20826 In conclusion, the radiotherapy causes reduction of bone matrix and modifies the morphology of bone channels network. Microsc. Res. Tech., 2010. © 2010 Wiley-Liss, Inc. (Source: Microscopy Research and Technique) Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3287474 Thu, 18 Feb 2010 00:00:00 +0100 3287474 Characterization of nanoscale transformations in polyelectrolyte multilayers fabricated from plasmid DNA using laser scanning confocal microscopy in combination with atomic force microscopy http://www.medworm.com/index.php?rid=3269593&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20830 Laser scanning confocal microscopy (LSCM) and atomic force microscopy (AFM) were used to characterize changes in nanoscale structure that occur when ultrathin polyelectrolyte multilayers (PEMs) are incubated in aqueous media. The PEMs investigated here were fabricated by the deposition of alternating layers of plasmid DNA and a hydrolytically degradable polyamine onto a precursor film composed of alternating layers of linear poly(ethylene imine) (LPEI) and sodium poly(styrene sulfonate) (SPS). Past studies of these materials in the context of gene delivery revealed transformations from a morphology that is smooth and uniform to one characterized by the formation of nanometer-scale particulate structures. We demonstrate that in-plane registration of LSCM and AFM images acquired from the sam... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3269593 Sat, 13 Feb 2010 00:00:00 +0100 3269593 Doxazosin treatment alters stromal cell behavior and increases elastic system fibers deposition in rat prostate http://www.medworm.com/index.php?rid=3269594&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20828 Doxazosin (DOX), an [alpha]-adrenoceptor antagonist, induces the relaxation of smooth muscle cell tonus and reduces the clinical symptoms of benign prostatic hyperplasia (BPH). However, the effects of DOX in the prostate stromal microenvironment are not fully known. In a previous study, we showed that DOX treatment for 30 days increased deposition of collagen fibers in the three rat prostatic lobes. Herein, we investigated the effects of DOX on stromal cell ultrastructure and elastic fiber deposition. Adult Wistar rats were treated with DOX (25 mg/kg/day); and the ventral, dorsal, and anterior prostates were excised at 30 days of treatment. The prostatic lobes were submitted to histochemical and stereological-morphometric analyze and transmission electron microscopy (TEM). Histochemical st... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3269594 Fri, 12 Feb 2010 00:00:00 +0100 3269594 Image processing techniques in a preliminary morphometric characterization of corneal epithelium by scanning electron microscopy http://www.medworm.com/index.php?rid=3258682&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20833 Several processing techniques of digital images allowed us to quantify the percentage of cell surface covered by microprojections (microvilli or microplicae) (SCM), the adhesion between epithelial cells by the parameter intercellular junctions (IJ), the size (cell area), shape (cell shape) and shade (cell shade) of cells on the corneal epithelium of nine rabbits. The data were analyzed and the epithelial cells were classified into three groups by cluster analysis. Assuming the representativeness of the sample, our findings suggest that for a normal corneal epithelium, 80% of the cells could show SCM >41%, and IJ >0.98 (being one a cell to cell junction without disruptions). Standard deviations of cell shade lower than 21 gray levels could indicate a tendency to lose the cell shade mosaic. ... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3258682 Wed, 10 Feb 2010 00:00:00 +0100 3258682 Erratum: Gomez F. and Romero E., a model for predicting pathologist's velocity profiles when navigating virtual slides. Microscopy research and technique 73:85-98. http://www.medworm.com/index.php?rid=3258687&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20799 No abstract.&#13; (Source: Microscopy Research and Technique) Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3258687 Tue, 09 Feb 2010 00:00:00 +0100 3258687 Leptin-like immunoreactivity in the muscle of juvenile sea bass (Dicentrarchus labrax) http://www.medworm.com/index.php?rid=3258686&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20822 In this study, we detected and localized leptin immunoreactivity to the muscle of early juvenile sea bass (Dicentrarchus labrax) by Western blot analysis and immunohistochemistry. A leptin immunopositive band with a molecular weight of [sim]16 kDa, corresponding to mammalian leptin, was identified in trunk skeletal muscle homogenate. Furthermore, leptin immunopositive cells were detected in the endomysium of skeletal muscular fibers. These cells showed immunostained cytoplasmic granules and roundish and oval nuclei. The most intense immunostaining was observed in the endomysial space among the superficial red muscular fibers of the trunk. These findings suggest that in early juvenile sea bass, leptin is mostly produced by skeletal muscles. Therefore, during the developmental stage lacking ... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3258686 Tue, 09 Feb 2010 00:00:00 +0100 3258686 Polylactide nanoparticles containing stably incorporated cyanine dyes for in vitro and in vivo imaging applications http://www.medworm.com/index.php?rid=3258685&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20824 Stably incorporating fluorescent molecules to polymeric nanoparticles (NPs) or micelles can facilitate the prolonged tracking of these drug-delivery vehicles in vitro and in vivo. However, incorporation of fluorescent molecules, usually charged and thereby water-soluble, through the encapsulation strategy to hydrophobic polymer matrices is challenging. The encapsulated fluorescent agents are also subject to rapid release when the polymeric NPs are exposed to biological media. To address this issue, we developed Cy5-conjugated polylactide (Cy5-PLA) NPs through Cy5/(BDI)ZnN(TMS)2 [(BDI) = 2-((2,6-diisopropylphenyl)amido)-4-((2,6-diisopropylphenyl)-imino)-2-pentene]-mediated ring-opening polymerization of lactide (LA) followed by nanoprecipitation. This process allows for covalent conjugation... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3258685 Tue, 09 Feb 2010 00:00:00 +0100 3258685 The administration of nonmetabolizable glucose analogues fails to suppress the development of glycogen autophagy in newborn rat hepatocytes http://www.medworm.com/index.php?rid=3258684&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20825 The effects of parenteral administration of glucose, 3-methylglucose (3MG), or 2-deoxyglucose (2DG) on the glycogen autophagy were studied in the newborn rat liver using electron microscopy and biochemical methods. The administration of glucose resulted in hyperglycemia and prevented the mobilization of hepatocytic glycogen. It also prevented the development of autophagic vacuoles in general and inhibited the glycogen-degrading activity of acid [alpha]-1,4-glucosidase. The nonphosphorylated and not further metabolized glucose analog 3MG also produced hyperglycemia, but increased acid glucosidase. Pretreating the newborns with the [beta]-adrenergic blocker propranolol inhibited the effects of 3MG. The phosphorylated but not fully metabolized glucose analog 2DG produced similar effects. The ... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3258684 Tue, 09 Feb 2010 00:00:00 +0100 3258684 Microhardness and SEM after CO2 laser irradiation or fluoride treatment in human and bovine enamel http://www.medworm.com/index.php?rid=3258683&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20827 It remains uncertain as to whether or not CO2 laser is able to hinder demineralization of enamel. The possibility to use bovine instead of human teeth on anticariogenic studies with laser has not yet been determined.To compare the ability of CO2 laser and fluoride to inhibit caries-like lesions in human enamel and to test whether a similar pattern of response would hold for bovine enamel.Ninety-six enamel slabs (2 × 2 × 4 mm) (48 from bovine and 48 from human teeth) were randomly distributed according to surface treatment (n = 12): CO2 laser, 5% sodium fluoride varnish (FV), 1.23% acidulated phosphate fluoride (APF) gel, or no treatment (control). Specimens were subjected to a 14-day in vitro cariogenic challenge. Microhardness (SMH) was measured at 30 [mu]m from the surface. For ultrast... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3258683 Tue, 09 Feb 2010 00:00:00 +0100 3258683 The hepatocytes of the brown trout (Salmo trutta fario): A stereological study of some cytoplasmic components with the breeding cycle http://www.medworm.com/index.php?rid=3239289&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20820 Sex differences exist in fish hepatocytes, but studies for characterizing their cytology throughout the breeding cycle are still scarce; suggesting changes, but most lacking quantitative data. To address this limitation, to complement baseline data generated from the brown trout model, and to prove that sex-specific seasonal changes exist, we made an unbiased stereological evaluation of the hepatocytic cytoplasm. Unprecedentedly for fish liver, the stereological design was exempt from model (biased) assumptions. Five (3 years old) animals per sex were studied in endogenous vitellogenesis, exogenous vitellogenesis, and spawning season end. Liver pieces for analysis were systematically sampled. Stereology was done in transmission electron microscopy (TEM) micrographs. Primary data generated ... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3239289 Thu, 04 Feb 2010 00:00:00 +0100 3239289 Influence of acceleration voltage on scanning electron microscopy of human blood platelets http://www.medworm.com/index.php?rid=3188956&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20778 Scanning electron microscopy (SEM) is used to view a variety of surface structures, molecules, or nanoparticles of different materials, ranging from metals, dental and medical instruments, and chemistry (e.g. polymer analysis) to biological material. Traditionally, the operating conditions of the SEM are very important in the material sciences, particularly the acceleration voltage. However, in biological sciences, it is not typically seen as an important parameter. Acceleration voltage allows electrons to penetrate the sample; thus, the higher the acceleration voltage the more penetration into the sample will occur. As a result, ultrastructural information from deeper layers will interfere with the actual surface morphology that is seen. Therefore, ultimately, if acceleration voltage is l... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3188956 Tue, 19 Jan 2010 00:00:00 +0100 3188956 Ultramorphological characteristics of immature stages of Chrysomya albiceps (Wiedemann 1819) (Diptera: Calliphoridae), a fly specie of forensic importance http://www.medworm.com/index.php?rid=3156340&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20819 Forensic entomology is an area of science that serves as a tool in crime scene investigations. Usually, flies are the first insects to reach a dead body and can oviposit just a few hours after arrival. Therefore the knowledge of immature stages is essential for correct identification of the species found on corpses. Scanning electron microscopy gives detailed information about morphological characters helping to identify the immature forms and consequently serves as a tool in crime scene investigations. C. albiceps is a very important fly for forensic entomologists because its larvae are almost always present on a dead body and it is facultative predators and therefore can alter the composition of species present at the carcass. The aim of this study is to identify eggs, larvae, and pupari... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3156340 Sat, 09 Jan 2010 00:00:00 +0100 3156340 Modified paraffin wax for improvement of histological analysis efficiency http://www.medworm.com/index.php?rid=3156341&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20818 We describe a modified paraffin wax that can improve the histological analysis efficiency of natural tissue, composed of paraffin and ethylene vinyl acetate (EVA) resin (0, 3, 5, and 10 wt %). Softening temperature of the paraffin/EVA media was similar to that of paraffin (50-60°C). The paraffin/EVA media dissolved completely in xylene after 30 min at 50°C. Physical properties such as the amount of load under the same compressive displacement, elastic recovery, and crystal intensity increased with increased EVA content. EVA medium (5 wt %) was regarded as an optimal composition, based on the sectioning efficiency measured by the numbers of unimpaired sectioned slices, amount of load under the same compressive displacement, and elastic recovery test. Based on the staining test of sectione... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3156341 Fri, 08 Jan 2010 00:00:00 +0100 3156341 Integrated methodology for the evaluation of cleaning effectiveness in two Russian icons (16th-17th centuries) http://www.medworm.com/index.php?rid=3126494&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20817 This article covers a methodology for evaluating the effectiveness of cleaning two Russian icons. The icons belong to a group of five from the same iconographic school, dating from the 16th to 17th centuries. An integrated and complementary approach to varnish and overpaint removal involved microscopic techniques (optical and scanning electron microscopy) and colorimetry (CIE L*a*b* system). The materials and techniques used in these icons have been characterized previously. Cleaning revealed extensive overpainting that had not only dramatically changed the original appearance, but also the meaning and attribution of one of the two icons. The analyses carried out were useful in determining the extent of the overpainting and led to a better assessment of the results and effectiveness of the... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3126494 Tue, 29 Dec 2009 00:00:00 +0100 3126494 Physical attachment of fluorescent protein particles to atomic force microscopy probes in aqueous media: Implications for surface pH, fluorescence, and mechanical properties studies http://www.medworm.com/index.php?rid=3118704&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20816 Transfer of a fluorescently labeled protein particle from a surface to a microsized scanning probe has been induced by repetitive scanning in aqueous medium. The so-attached particle can in turn act as a probing tool to study particle-substrate and particle-particle interactions. Attachment of the fluorescent particle occurs at the apical region of an atomic force microscope (AFM) cantilever tip and it endures repetitive loading-unloading cycles against the sample surface. Fluorescence microscopy has been used to address the exact location of the attached particle in the cantilever and to identify the moment when the particle contacts the sample. Moreover, we have observed that fluorescence intensity at the contact point is lower when the probing particle contacts another fluorescent parti... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3118704 Thu, 24 Dec 2009 00:00:00 +0100 3118704 Infection of the Red Palm Weevil (Rhynchophorus ferrugineus) by the entomopathogenic fungus Beauveria bassiana: A SEM study http://www.medworm.com/index.php?rid=3104859&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20812 The Red Palm Weevil (Rhynchophorus ferrugineus) is a devastating pest of palms in the Mediterranean, Middle East, and Eastern countries. No effective control measures are available. R. ferrugineus has been found naturally infected by the entomopathogenic fungus Beauveria bassiana, but its infection process in this host is unknown. We have studied the infection of R. ferrugineus larvae and adults by B. bassiana using dry conidia and conidia suspensions using scanning electron microscopy (SEM). In early stages, SEM revealed acquisition of B. bassiana conidia by cuticle ornamentation in legs, antennae, and elytra of R. ferrugineus adults. Subsequently, conidia germinated and frequent episodes of hyphal/conidial fusion were found. Appressoria, signs of adhesion and cuticle degradation led to p... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3104859 Sat, 19 Dec 2009 00:00:00 +0100 3104859 The cathepsin K inhibitor AAE581 induces morphological changes in osteoclasts of treated patients http://www.medworm.com/index.php?rid=3104862&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20813 Inhibitors of Cathepsin K (Cat-K) are recognized as an interesting way to inhibit osteoclast (OC) activity. OCs from patients treated with the anticathepsin-K inhibitor AAE581 (balicatib) were found enlarged. They contained numerous vacuoles filled with tartrate resistant acid phosphatase (TRAcP), an intracellular enzyme that terminates the degradation of collagen internalized in OC transcytotic vesicles. In a phase 2 clinical study, 675 patients with postmenopausal osteoporosis received the Cat-K inhibitor AAE581 at 0, 5, 10, 25, or 50 mg/D during 1 year. Eleven patients had a transiliac bone biopsy, studied undecalcified. Histoenzymatic detection of TRAcP was used to identify and count OC number. The histomorphometrist was not aware of the randomization of patients at the time of analysi... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3104862 Fri, 18 Dec 2009 00:00:00 +0100 3104862 Curcumin protects against hepatic and renal injuries mediated by inducible nitric oxide synthase during selenium-induced toxicity in Wistar rats http://www.medworm.com/index.php?rid=3104861&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20802 The aim of this study is to evaluate the effect of curcumin in protecting against selenium-induced toxicity in liver and kidney of Wistar rats. Light microscopy evaluation of selenium alone administered rats showed liver to be infiltrated with mononuclear cells, vacuolation, necrosis, and pronounced degeneration. Control liver sections showed a regular morphology of parenchymal cells with intact hepatocytes and sinusoids. Kidney from selenium alone administered rats showed vacuolar degeneration changes in the epithelial cells, cellular proliferation with fibrosis, thickening of capillary walls, and glomerular tuft atrophy. Such changes were also observed in rats administered with selenium and curcumin simultaneously and rats administered first with selenium and then curcumin 24 h later. In... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3104861 Fri, 18 Dec 2009 00:00:00 +0100 3104861 A reproducible technique for specific labeling of antigens using preformed fluorescent molecular IgG-F(ab[prime])2 complexes from primary antibodies of the same species http://www.medworm.com/index.php?rid=3104860&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20803 In this study, we describe how preformed complexes of primary and secondary labeled antibodies can be used in such circumstances. In this situation, the first antigen is labeled using the conventional indirect method followed by incubation with the preformed primary-secondary antibody complex against the second antigen. To prevent unbound secondary antibody from binding the indirectly-labeled antibodies, resulting in a false positive, we quenched excess secondary antibody with nonimmune murine serum from the species of the primary antibody. Before the formation of the preformed complex, the optimum dilution of both primary and secondary antibodies was determined. Once these concentrations were established, the concentration of nonimmune murine serum required to quench excess unbound second... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3104860 Fri, 18 Dec 2009 00:00:00 +0100 3104860 Technical advances in the sectioning of dental tissue and of on-section cross-linked collagen detection in mineralized teeth http://www.medworm.com/index.php?rid=3047926&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20815 This report offers advances in sectioning mineralized dental tissues and allows the application of immunofluorescence not only for on-section protein detection but importantly for detecting cross-linked fibrous collagens in both soft and mineralized tissue sections. Microsc. Res. Tech., 2010. © 2009 Wiley-Liss, Inc. (Source: Microscopy Research and Technique) Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3047926 Wed, 02 Dec 2009 00:00:00 +0100 3047926 Use of direct fluorescence labeling and confocal microscopy to determine the biodistribution of two protein therapeutics, Cerezyme® and Ceredase® http://www.medworm.com/index.php?rid=3047930&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20810 Efficient targeting of therapeutic reagents to tissues and cell types of interest is critical to achieving therapeutic efficacy and avoiding unwanted side effects due to offtarget uptake. To increase assay efficiency and reduce the number of animals used per experiment during preclinical development, we used a combination of direct fluorescence labeling and confocal microscopy to simultaneously examine the biodistribution of two therapeutic proteins, Cerezyme® and Ceredase®, in the same animals. We show that the fluorescent tags do not interfere with protein uptake and localization. We are able to detect Cerezyme and Ceredase in intact cells and organs and demonstrate colocalization within target cells using confocal microscopy. In addition, the relative amount of protein internalized by... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3047930 Tue, 01 Dec 2009 00:00:00 +0100 3047930 Detection of lead in Zea mays by dual-energy X-ray microtomography at the SYRMEP beamline of the ELETTRA synchrotron and by atomic absorption spectroscopy http://www.medworm.com/index.php?rid=3047929&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20805 This study is related to the application of the X-ray dual-energy microradiography technique together with the atomic absorption spectroscopy (AAS) for the detection of lead on Zea mays stem, ear, root, and leaf samples. To highlight the places with lead intake, the planar radiographs taken with monochromatic X-ray radiation in absorption regime with photon energy below and above the absorption edge of a given chemical element, respectively, are analyzed and processed. To recognize the biological structures involved in the intake, the dual-energy images with the lead signal have been compared with the optical images of the same Z. mays stem. The ear, stem, root, and leaf samples have also been analyzed with the AAS technique to measure the exact amount of the hyperaccumulated lead. The AAS... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3047929 Tue, 01 Dec 2009 00:00:00 +0100 3047929 Enameloid microstructure of the serrated cutting edges in certain fossil carcharhiniform and lamniform sharks http://www.medworm.com/index.php?rid=3047928&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20811 This study uses scanning electron microscopy to investigate the orientation of the parallel enameloid bundles in the area of the serrated cutting edges in certain fossil elasmobranchs. The examined teeth come from two Upper Cretaceous Squalicorax species and the Upper Miocene carcharhiniforms Galeocerdo sp., Carcharhinus sp., and Hemipristis serra. The parallel bundles are revealed by surface etching, which removes the superficial shiny-layered enameloid. In the teeth of Squalicorax, the bundles around the cutting edge bend once, before they reach the serrations. The studied carcharhiniform species show a more complicated pattern with a change of parallel bundle course inside the serrations. H. serra teeth do not display the first bending of the bundles, whereas it was present in the other... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3047928 Tue, 01 Dec 2009 00:00:00 +0100 3047928 A single pretreatment by zoledronic acid converts metastases from osteolytic to osteoblastic in the rat http://www.medworm.com/index.php?rid=3047927&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20814 Bone metastases are severe complications of cancers associated with increased morbidity, pain, risk fracture, and reduced life span for patients. Bisphosphonates emerged as a relief treatment in bone metastases. A single dose of zoledronic acid (78 [mu]g/kg) was injected into six Copenhagen rats 4 days before receiving an intraosseous inoculation of metastatic anaplastic tumor of lymph node and lung cell (MLL) prostate cancer cells. Rat femurs were analyzed for changes by microCT and histomorphometry; trabecular volume, trabecular characteristics, osteoid parameters, osteoblastic surfaces, and osteoclast number were measured. Values were compared to a group of SHAM animals, a group of SHAM animals having received zoledronic acid and animals inoculated with MLL cells. All rats were euthaniz... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3047927 Tue, 01 Dec 2009 00:00:00 +0100 3047927 Surfing the wave, cycle, life history, and genes/proteins expressed by testicular germ cells. Part 1: Background to spermatogenesis, spermatogonia, and spermatocytes http://www.medworm.com/index.php?rid=3029587&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20783 Spermatogenesis, a study of germ cell development, is a long, orderly, and well-defined process occurring in seminiferous tubules of the testis. It is a temporal event whereby undifferentiated spermatogonial germ cells evolve into maturing spermatozoa over a period of several weeks. Spermatogenesis is characterized by three specific functional phases: proliferation, meiosis, and differentiation, and it involves spermatogonia, spermatocytes, and spermatids. Germ cells at steps of development form various cellular associations or stages, with 6, 12, and 14 specific stages being identified in human, mouse, and rat, respectively. The stages evolve over time in a given area of the seminiferous tubule forming a cycle of the seminiferous epithelium that has a well-defined duration for a given spe... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3029587 Thu, 26 Nov 2009 00:00:00 +0100 3029587 A new bone vascular perfusion compound for the simultaneous analysis of bone and vasculature http://www.medworm.com/index.php?rid=3029597&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20807 In this study, we present a new barium sulfate (BaSO4) radiopaque vascular perfusion compound for performing postmortem microangiography with single-beam microcomputed tomography (microCT), which allows for such simultaneous analysis. This compound differs from currently available contrast mediums due to (1) the high weight-to-volume ratio of BaSO4 achieved, (2) small BaSO4 aggregate size ( (Source: Microscopy Research and Technique) Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3029597 Wed, 25 Nov 2009 00:00:00 +0100 3029597 Evaluation of the smear layer and hybrid layer in noncarious and carious dentin prepared by air abrasion system and diamond tips http://www.medworm.com/index.php?rid=3029596&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20798 To analyze the smear layer and the hybrid layer in noncarious and carious dentin prepared by different cutting instruments and restored with composite resin.Cavities were randomly prepared in 160 specimens (noncarious and artificial carious dentin) by high-speed diamond tips (KG Sorensen 1013), air abrasion system (Prepstart, Danville Engineering), ultrasonic tip (CVDentus 8.3231-1), and ultrasonic tip associated with ultrasonic cavitation by water for 10 s. Half of the cavities in each group were conditioned with 37% phosphoric acid for 15 s. The amount of smear layer and dentinal tubules present were analyzed using scanning electron microscopy and graded from 0 to 3. Cavities were prepared in another 20 noncarious specimens and 20 carious specimens and restored with adhesive composite re... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3029596 Wed, 25 Nov 2009 00:00:00 +0100 3029596 MRT letter: Auto-fluorescence by human alveolar macrophages after in vitro exposure to air pollution particles http://www.medworm.com/index.php?rid=3029595&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20804 Macrophages from smokers demonstrate an increased auto-fluorescence. Similarly, auto-fluorescence follows in vitro exposure of macrophages to cigarette smoke condensate (i.e., the particulate fraction of cigarette smoke). The composition of particles in cigarette smoke can be comparable to air pollution particles. We tested the postulate that macrophages exposed to air pollution particles could demonstrate auto-fluorescence. Healthy nonsmoking and healthy smoking volunteers (both 18-40 years of age) underwent fiberoptic bronchoscopy with bronchoalveolar lavage and alveolar macrophages isolated. Macrophages were incubated at 37°C in 5% CO2 with either PBS or 100 [mu]g/mL particle for both 1 and 24 h. Particles included a residual oil fly ash, Mt. St. Helens volcanic ash, and ambient air pa... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3029595 Wed, 25 Nov 2009 00:00:00 +0100 3029595 Flexible and stable optical parametric oscillator based laser system for coherent anti-Stokes Raman scattering microscopy http://www.medworm.com/index.php?rid=3029594&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20806 The characteristics of a stable and flexible laser system based on a synchronously pumped optical parametric oscillator (OPO) is presented. This OPO can offer very stable operation with both [sim]1 ps and [sim]300 fs outputs over a broad wavelength range, i.e., 920-1200 nm. Combining the pump tuning with the OPO tuning, a total Raman range of 1900-5500 cm-1 is accessible. For maximum spectral sensitivity, the CARS microsope based on the ps laser system is presented in detail. The lateral resolution of the microscope is diffraction limited to be about 390 nm. Fast wavelength switching (sub-second) between two Raman vibrational frequencies, i.e., 2848 cm-1 for C[bond]H aliphatic vibrations and 3035 cm-1 for C[bond]H aromatic vibrations is presented as an example, although this also extends t... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3029594 Wed, 25 Nov 2009 00:00:00 +0100 3029594 Oogenesis and programmed cell death of nurse cells in the endoparasitoid, Pteromalus puparum http://www.medworm.com/index.php?rid=3029593&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20808 In this study, we describe the features of oogenesis in the endoparasitoid, Pteromalus puparum, as well as the distinct type of programmed cell death of the nurse cells through conventional light and fluorescent markers for apoptosis and immunofluorescent analysis. Oogenesis in this endoparasitoid is divided into five stages, of which stages 1-2, 3-4, and 5 are corresponding to previtellogenic growth, yolk uptake, and the formation of egg envelopes, respectively. From these studies, we demonstrate two critical events, which are vitellogenin absorption and rapid transfer of nurse cell content, resulting in remarkable increase in the volume of oocytes during oogenesis in this endoparasitoid. Vitellogenin absorption initiates in the oocyte of early stage 3, and bulk transfer of nurse cell con... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3029593 Wed, 25 Nov 2009 00:00:00 +0100 3029593 Surfing the wave, cycle, life history, and genes/proteins expressed by testicular germ cells. Part 3: Developmental changes in spermatid flagellum and cytoplasmic droplet and interaction of sperm with the zona pellucida and egg plasma membrane http://www.medworm.com/index.php?rid=3029592&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20784 Spermiogenesis constitutes the steps involved in the metamorphosis of spermatids into spermatozoa. It involves modification of several organelles in addition to the formation of several structures including the flagellum and cytoplasmic droplet. The flagellum is composed of a neck region and middle, principal, and end pieces. The axoneme composed of nine outer microtubular doublets circularly arranged to form a cylinder around a central pair of microtubules is present throughout the flagellum. The middle and principal pieces each contain specific components such as the mitochondrial sheath and fibrous sheath, respectively, while outer dense fibers are common to both. A plethora of proteins are constituents of each of these structures, with each playing key roles in functions related to the... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3029592 Wed, 25 Nov 2009 00:00:00 +0100 3029592 Surfing the wave, cycle, life history, and genes/proteins expressed by testicular germ cells. Part 4: Intercellular bridges, mitochondria, nuclear envelope, apoptosis, ubiquitination, membrane/voltage-gated channels, methylation/acetylation, and transcription factors http://www.medworm.com/index.php?rid=3029591&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20785 As germ cells divide and differentiate from spermatogonia to spermatozoa, they share a number of structural and functional features that are common to all generations of germ cells and these features are discussed herein. Germ cells are linked to one another by large intercellular bridges which serve to move molecules and even large organelles from the cytoplasm of one cell to another. Mitochondria take on different shapes and features and topographical arrangements to accommodate their specific needs during spermatogenesis. The nuclear envelope and pore complex also undergo extensive modifications concomitant with the development of germ cell generations. Apoptosis is an event that is normally triggered by germ cells and involves many proteins. It occurs to limit the germ cell pool and ac... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3029591 Wed, 25 Nov 2009 00:00:00 +0100 3029591 Surfing the wave, cycle, life history, and genes/proteins expressed by testicular germ cells. Part 5: Intercellular junctions and contacts between germs cells and Sertoli cells and their regulatory interactions, testicular cholesterol, and genes/proteins associated with more than one germ cell generation http://www.medworm.com/index.php?rid=3029590&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20786 In the testis, cell adhesion and junctional molecules permit specific interactions and intracellular communication between germ and Sertoli cells and apposed Sertoli cells. Among the many adhesion family of proteins, NCAM, nectin and nectin-like, catenins, and cadherens will be discussed, along with gap junctions between germ and Sertoli cells and the many members of the connexin family. The blood-testis barrier separates the haploid spermatids from blood borne elements. In the barrier, the intercellular junctions consist of many proteins such as occludin, tricellulin, and claudins. Changes in the expression of cell adhesion molecules are also an essential part of the mechanism that allows germ cells to move from the basal compartment of the seminiferous tubule to the adluminal compartment... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3029590 Wed, 25 Nov 2009 00:00:00 +0100 3029590 Surfing the wave, cycle, life history, and genes/proteins expressed by testicular germ cells. Part 2: Changes in spermatid organelles associated with development of spermatozoa http://www.medworm.com/index.php?rid=3029589&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20787 Spermiogenesis is a long process whereby haploid spermatids derived from the meiotic divisions of spermatocytes undergo metamorphosis into spermatozoa. It is subdivided into distinct steps with 19 being identified in rats, 16 in mouse and 8 in humans. Spermiogenesis extends over 22.7 days in rats and 21.6 days in humans. In this part, we review several key events that take place during the development of spermatids from a structural and functional point of view. During early spermiogenesis, the Golgi apparatus forms the acrosome, a lysosome-like membrane bound organelle involved in fertilization. The endoplasmic reticulum undergoes several topographical and structural modifications including the formation of the radial body and annulate lamellae. The chromatoid body is fully developed and ... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3029589 Wed, 25 Nov 2009 00:00:00 +0100 3029589 Mirror lenses in light microscopy - Theoretical considerations and practical implications http://www.medworm.com/index.php?rid=3029588&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20809 Various types of mirror lenses were developed some decades ago designed for several special tasks in microscopy. When compared with high-end glass lenses, mirror lenses lead to an extraordinary image quality because of their supraapochromatic color fidelity and their high planarity; the working distances are significantly longer, and in most cases, the depth of field and resolution are higher than in concurrent glass lenses. In this respect, the microscopic advantages of mirror lenses seem to be comparable with the advantages of mirror telescopes in astronomy; for in observations of celestial bodies, mirror telescopes lead to better results than refractors based on glass lenses. When mirror lenses are available in light microscopy, not only common illumination modes can be carried out in t... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3029588 Wed, 25 Nov 2009 00:00:00 +0100 3029588 Optimized time-gated generalized polarization imaging of Laurdan and di-4-ANEPPDHQ for membrane order image contrast enhancement http://www.medworm.com/index.php?rid=3022929&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20801 The membrane dyes Laurdan and di-4-ANEPPDHQ can be used to image membrane order due to a spectral blue-shift in the fluorescence emission between the liquid-ordered and liquid-disordered phases. These images typically take the form of a normalized intensity ratio image known as a generalized polarization (GP) plot. Here, we exploit the known excited state photophysics and time-resolved data acquisition via time-correlated single-photon counting (TCSPC) to demonstrate GP contrast enhancement for these two probes of 7 and 31%, respectively. This improvement in image contrast enhancement will be invaluable when studying the role of lipid rafts in fixed and live cell systems. Microsc. Res. Tech. 2009. © 2009 Wiley-Liss, Inc. (Source: Microscopy Research and Technique) Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3022929 Tue, 24 Nov 2009 00:00:00 +0100 3022929 Evaluation of critical size defects of mouse calvarial bone: An organ culture study http://www.medworm.com/index.php?rid=3022936&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20792 Mouse calvarial organ culture has been used widely for the study of bone biology. The purpose of this study was to evaluate the healing potential of neonatal mouse parietal defects in different culture media. The critical size defect (CSD) was also investigated. The parietal bones of neonatal mice were used. Full-thickness, 0.8-mm circular defects were created through the bones from one litter of mice. The bones were divided into three groups: Dulbecco's Modified Eagle Medium (DMEM) group, DMEM/osteogenic medium (OM) group, and OM group. Cultures were analyzed with microcomputed-tomography, dissecting-microscope, phase-contrast-microscope, Von Kossa stain, scanning-electron-microscopy, and energy-dispersive-X-ray. Continuous bone formation of parietal bones was observed in all groups. Defe... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3022936 Mon, 23 Nov 2009 00:00:00 +0100 3022936 In situ hybridization and immunofluorescence on resin-embedded tissue to identify the components of Nissl substance http://www.medworm.com/index.php?rid=3022935&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20794 It is not clear whether the Nissl substance is present at the axon hillock. To clarify this gap in knowledge, we conducted in situ hybridization (ISH) on mouse brain tissue using 30-[mu]m cryostat and 1-3-[mu]m acrylic resin sections. Cryostat and rehydrated resin sections were exposed to digoxygenin-labeled glutamic acid decarboxylase 1 sense and antisense riboprobes. Consecutive sections from tissue embedded in resin were subjected to the ribosomal protein L26 primary antibody to determine the distribution of the ribo/polysomes. ISH results from the antisense riboprobe in both cryostat and resin-embedded tissue sections demonstrated an abundance of message in the neurons from the substantia nigra pars reticulate. In addition, the resin sections demonstrated hybridization signal in the ax... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3022935 Mon, 23 Nov 2009 00:00:00 +0100 3022935 Sensilla on the antennae, legs and ovipositor of Spathius agrili Yang (Hymenoptera: Braconidae), a parasitoid of the emerald ash borer Agrilus planipennis Fairmaire (Coleoptera: Buprestidae) http://www.medworm.com/index.php?rid=3022934&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20795 To investigate the host-finding mechanisms of Spathius agrili Yang, a parasitoid of the emerald ash borer, Agrilus planipennis Fairmaire, a highly concealed insect pest of Fraxinus spp. The morphology and ultrastructure of sensilla on the antennae and legs of both sexes and on the ovipositors and ovipositor sheaths of females were observed using SEM. Results showed that abundant sensilla trichodea (ST) are distributed on the antennae of both females and males, with the sharp trichoid with a socket at the base being the most numerous sensillum. There are 6-7 sensilla placodea on each segment of the flagellum, arranged lengthways in parallel rows. There are very few sensilla styloconica on the female antennae. The tibiae have mostly sensilla chaetica, whereas three types of ST and the relati... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3022934 Mon, 23 Nov 2009 00:00:00 +0100 3022934 Development of an optimized protocol for studying the interaction of filamentous bacteriophage with mammalian cells by fluorescence microscopy http://www.medworm.com/index.php?rid=3022933&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20793 Filamentous bacteriophage has been proposed as a vehicle that can carry and deliver therapeutics into mammalian cells for disease treatment, thus a protocol for imaging phage-cell interaction is essential. Because high signal intensity is necessary to study the mechanism of interaction between filamentous bacteriophage and mammalian cells, it is important to optimize the procedure for fluorescence labeling of phage in order to understand such interaction. Here, we describe a procedure that gives intense fluorescence labeling and can show interactions between fd-tet bacteriophage selected from phage libraries and mammalian cells (SKBR-3 and MCF-10A). The indirect labeling of phage with dye-conjugated antibody and cytoskeleton associated proteins was significantly enhanced in the presence of... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3022933 Mon, 23 Nov 2009 00:00:00 +0100 3022933 Microstructure and mineral composition of dental enamel of permanent and deciduous teeth http://www.medworm.com/index.php?rid=3022932&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20796 Conclusions: The primary enamel structure showed a lower level of Ca and P, thinner thickness and higher numerical density of rods. Microsc. Res. Tech., 2009. © 2009 Wiley-Liss, Inc. (Source: Microscopy Research and Technique) Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3022932 Mon, 23 Nov 2009 00:00:00 +0100 3022932 Aberrant distribution of junctional complex components in retinoic acid receptor alpha-deficient mice http://www.medworm.com/index.php?rid=3022931&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20797 In this study, we investigated whether defective retinoid signaling involved at least in part, disrupted cell-cell interactions. Hypertonic fixation approaches revealed defects in the integrity of the Sertoli-cell barrier in the tubules of RAR[alpha]-deficient testes. Dye transfer experiments further revealed that coupling between cells from the basal to adluminal compartments was aberrant. There were also differences in the expression of several known retinoic acid (RA)-responsive genes encoding structural components of tight junctions and gap junctions. Immunostaining demonstrated a delay in the incorporation of zonula occludens (ZO-1), a peripheral component protein of tight junctions, into the Sertoli cell tight junctions. Markedly reduced expression of connexin-40 in mutant pachytene ... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3022931 Mon, 23 Nov 2009 00:00:00 +0100 3022931 Synaptic membrane proteins form stable microdomains in early endosomes http://www.medworm.com/index.php?rid=3022930&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20800 In the plasma membrane, membrane proteins are frequently organized in microdomains that are stabilized both by protein-protein and protein-lipid interactions, with the membrane lipid cholesterol being instrumental for microdomain stability. However, it is unclear whether such microdomains persist during endocytotic membrane trafficking. We used stimulated emission-depletion microscopy to investigate the domain structure of the endosomes. We developed a semiautomatic method for counting the individual domains, an approach that we have validated by immunoelectron microscopy. We found that in endosomes derived from neuroendocrine PC12 cells synaptophysin and several SNARE proteins are organized in microdomains. Cholesterol depletion by methyl-[beta]-cyclodextrin disintegrates most of the doma... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=3022930 Mon, 23 Nov 2009 00:00:00 +0100 3022930 Ultrastructural alterations in liver of mice exposed chronically and transgenerationally to aqueous extract of betel nut: Implications in betel nut-induced carcinogenesis http://www.medworm.com/index.php?rid=2901555&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20791 The aqueous extract of betel nut (AEBN) induces the formation of preneoplastic nodules in the liver of Swiss Albino mice and leads to increased predisposition to cancer when administered transgenerationally. The aim of this investigation was to elucidate the alterations in ultrastructure of subcellular organelles in the liver nodules using transmission electron microscopy and to determine whether these alterations have implications in AEBN-induced carcinogenesis. Male and female Swiss Albino mice were exposed to AEBN chronically and transgenerationally at a dose of 2 mg/mL in drinking water for 24 weeks. Extensive polymorphism was noted in nuclear shape and heterochromatin organization. Heterochromatin aggregation and marginalization were observed in the nuclei of chronically exposed mice,... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2901555 Fri, 16 Oct 2009 23:00:00 +0100 2901555 Quantitative analysis of spatial proteoglycan content in articular cartilage with Fourier transform infrared imaging spectroscopy: Critical evaluation of analysis methods and specificity of the parameters http://www.medworm.com/index.php?rid=2901558&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20789 To evaluate the specificity of the current Fourier transform infrared imaging spectroscopy (FT-IRIS) methods for the determination of depthwise proteoglycan (PG) content in articular cartilage (AC). In addition, curve fitting was applied to study whether the specificity of FT-IRIS parameters for PG determination could be improved.Two sample groups from the steer AC were prepared for the study (n = 8 samples/group). In the first group, chondroitinase ABC enzyme was used to degrade the PGs from the superficial cartilage, while the samples in the second group served as the controls. Samples were examined with FT-IRIS and analyzed using previously reported direct absorption spectrum techniques and multivariate methods and, in comparison, by curve fitting. Safranin O-stained sections were measu... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2901558 Thu, 15 Oct 2009 23:00:00 +0100 2901558 Chemical and morphological features of dental composite resin: Influence of light curing units and immersion media http://www.medworm.com/index.php?rid=2901557&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20769 The study evaluated the influence of light curing units and immersion media on superficial morphology and chemistry of the nanofilled composite resin Supreme XT (3M) through the EDX analysis and SEM evaluation. Light curing units with different power densities and mode of application used were XL 3000 (480 mW/cm2), Jet Lite 4000 Plus (1230mW/cm2), and Ultralume Led 5 (790 mW/cm2) and immersion media were artificial saliva, Coke®, tea and coffee, totaling 12 experimental groups. Specimens (10 mm × 2 mm) were immersed in each respective solution for 5 min, three times a day, during 60 days and stored in artificial saliva at 37°C ± 1°C between immersion periods. Topography and chemical analysis was qualitative.Groups immersed in artificial saliva, showed homogeneous degradation of matrix... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2901557 Thu, 15 Oct 2009 23:00:00 +0100 2901557 Development and neuronal dependence of cutaneous sensory nerve formations: Lessons from neurotrophins http://www.medworm.com/index.php?rid=2901556&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20790 Null mutations of genes from the NGF family of NTs and their receptors (NTRs) lead to loss/reduction of specific neurons in sensory ganglia; conversely, cutaneous overexpression of NTs results in skin hyperinnervation and increase or no changes in the number of sensory neurons innervating the skin. These neuronal changes are paralleled with loss of specific types of sensory nerve formations in the skin. Therefore, mice carrying mutations in NT or NTR genes represent an ideal model to identify the neuronal dependence of each type of cutaneous sensory nerve ending from a concrete subtype of sensory neuron, since the development, maintenance, and structural integrity of sensory nerve formations depend upon sensory neurons. Results obtained from these mouse strains suggest that TrkA positive n... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2901556 Thu, 15 Oct 2009 23:00:00 +0100 2901556 Progressive chorion morphology during egg development in Samia ricini (Donovan) http://www.medworm.com/index.php?rid=2834196&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20781 The egg of Samia ricini (Donovan), is oval or laterally flattened ellipsoid, freshly laid eggs are candid white while the chorion is colorless and semi-transparent. The surface of the chorion is covered with network patterns of polygons and their shapes are common in the whole surface region. The boundaries between polygons made ridges had distinct acropyles at three-cell junctions. The numbers of aeropyles are variable according to their structures both in the lateral flat and marginal regions. During the course of egg development, no significant structural changes were observed in either the polygonal structures or the overall morphology of the egg. However, the size of the aeropyles kept on changing as the egg matures. The aeropyle increases initially upto day-9 of egg development and t... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2834196 Fri, 25 Sep 2009 23:00:00 +0100 2834196 Green fluorescent protein as indicator of nonviral transient transfection efficiency in endometrial and testicular biopsies http://www.medworm.com/index.php?rid=2834200&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20779 In conclusion, nonviral methods are attractive for an improvement of the gene therapy and our protocol could provide useful indications for in vivo gene therapy applications. Microsc. Res. Tech., 2009. © 2009 Wiley-Liss, Inc. (Source: Microscopy Research and Technique) Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2834200 Thu, 24 Sep 2009 23:00:00 +0100 2834200 MRT letter: In situ observation method for microstructural changes of steel during hot deformation http://www.medworm.com/index.php?rid=2834199&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20780 We report on the result of an in situ method for observing microstructural changes during hot deformation. The observation of microstructural changes of steel at 1,473 K under tensile strain is demonstrated using the reported method. The development of deformed structures and the formation of a new grain boundary, which subsequently moved with increased strain, were clearly observed. The effectiveness of this method was confirmed by the results of several examples. © 2009 Wiley-Liss, Inc. (Source: Microscopy Research and Technique) Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2834199 Thu, 24 Sep 2009 23:00:00 +0100 2834199 Spermatozoon and its relationship with the ovarian lamellae in the internally inseminating catfish Trachelyopterus galeatus http://www.medworm.com/index.php?rid=2834198&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20782 We examined the spermatozoa and their relationship with the ovarian lamellae in the catfish Trachelyopterus galeatus by classical light microscopy, high-resolution light microscopy, and transmission electron microscopy. Trachelyopterus galeatus is an internally inseminating species the spermatozoon of which presented an elongated cylindrical head (12.3 ± 1.5 [mu]m), elongated midpiece (5.0 ± 0.7 [mu]m), and flagellum (23.9 ± 2.8 [mu]m). Fertilized eggs or embryos were not found in its ovaries. Spermatozeugmata were demonstrated for the first time in this species. At the ultrastructural level, the anterior region of the head was devoid of chromatin with its shape being rounded with a hyaline tip in frontal sections and flattened in sagittal sections. The proximal centriole and most of th... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2834198 Thu, 24 Sep 2009 23:00:00 +0100 2834198 Autophagosomal glycogen-degrading activity and its relationship to the general autophagic activity in newborn rat hepatocytes: The effects of parenteral glucose administration http://www.medworm.com/index.php?rid=2834197&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20788 The effects of the administration of parenteral glucose on the postnatal glycogen autophagic activity and its relationship to the general autophagic activity, were studied in newborn rat liver using electron microscopy and biochemical methods. Glucose abolished the normal postnatal hypoglycemia and preserved the hepatocytic hyaloplasmic glycogen to the levels of birth. It also inhibited the normal postnatal increase in the number and volume of autophagic vacuoles. Glucose especially decreased the rate of postnatal development of the glycogen-containing autophagic vacuoles. This decrease was greater than that of the autophagic vacuoles in general. In the control animals at the age of 6 h, the total volume of the glycogen-containing autophagic vacuoles accounted for 87% of the autophagic vac... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2834197 Thu, 24 Sep 2009 23:00:00 +0100 2834197 A novel approach for correlative light electron microscopy analysis http://www.medworm.com/index.php?rid=2757484&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20777 Correlative light and electron microscopy (CLEM) is a multimodal technique of increasing utilization in functional, biochemical, and molecular biology. CLEM attempts to combine multidimensional information from the complementary fluorescence light microscopy (FLM) and electron microscopy (EM) techniques to bridge the various resolution gaps. Within this approach the very same cell/structure/event observed at level can be analyzed as well by FLM and EM. Unfortunately, these studies turned out to be extremely time consuming and are not suitable for statistical relevant data. Here, we describe a new CLEM method based on a robust specimen preparation protocol, optimized for cryosections (Tokuyasu method) and on an innovative image processing toolbox for a novel type of multimodal analysis. Mai... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2757484 Tue, 01 Sep 2009 23:00:00 +0100 2757484 Immunohistochemical characterization of TH13-L2 spinal ganglia neurons in sheep (Ovis aries) http://www.medworm.com/index.php?rid=2757497&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20764 In this study, some of the widely used neuronal markers, neurofilament 200 kDa (NF200), substance P (SP), calcitonin gene-related peptide (CGRP) and isolectin B4 (IB4), were employed to characterize neuronal nitric oxide synthase (nNOS)-immunoreactivity (-IR) in sheep (Ovis aries) SG (Th13-L2) neurons. The majority of the SG neurons were IB4-labeled (79 ± 10%), followed by NF200- (45 ± 4%), NOS- (44 ± 10%), SP- (42 ± 5%) and CGRP-IR (35 ± 7%) neurons. The triple staining experiments showed that a higher percentage (75 ± 16%) of NOS-IR neurons bound both IB4 and CGRP, or both IB4 and SP (49 ± 6%). The IB4 marker showed an unexpected staining pattern; in fact, IB4-labeled neurons largely colocalized with NF200, usually considered a marker of light SG neurons, and with CGRP and SP. For... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2757497 Mon, 31 Aug 2009 23:00:00 +0100 2757497 Histometric and histomorphologic comparison of combustion and ambustion using in vivo reflectance-confocal microscopy http://www.medworm.com/index.php?rid=2757496&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20770 Conclusions: RCM evaluates significant histomorphological differences in superficial burns caused by combustion and ambustion. The term "superficial burn" should consider the underlying cause and thus supplemented by the term "combustion" or "ambustion." Microsc. Res. Tech., 2009. © 2009 Wiley-Liss, Inc. (Source: Microscopy Research and Technique) Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2757496 Mon, 31 Aug 2009 23:00:00 +0100 2757496 Parallel deconvolution of large 3D images obtained by confocal laser scanning microscopy http://www.medworm.com/index.php?rid=2757495&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20773 Various deconvolution algorithms are often used for restoration of digital images. Image deconvolution is especially needed for the correction of three-dimensional images obtained by confocal laser scanning microscopy. Such images suffer from distortions, particularly in the Z dimension. As a result, reliable automatic segmentation of these images may be difficult or even impossible. Effective deconvolution algorithms are memory-intensive and time-consuming. In this work, we propose a parallel version of the well-known Richardson-Lucy deconvolution algorithm developed for a system with distributed memory and implemented with the use of Message Passing Interface (MPI). It enables significantly more rapid deconvolution of two-dimensional and three-dimensional images by efficiently splitting ... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2757495 Mon, 31 Aug 2009 23:00:00 +0100 2757495 The toxic effect of ketamine on SH-SY5Y neuroblastoma cell line and human neuron http://www.medworm.com/index.php?rid=2757494&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20774 Ketamine used as an injectable anesthetic in human and animal medicine is also a recreational drug used primarily by young adults often at all night dance parties in nightclubs. The percentage of ketamine users has grown very fast in the last 5 years worldwide. However, this leads to the serious question of the long-term adverse effects of ketamine on our nervous system, particularly the brain, because ketamine as an NMDA antagonist could cause neurons to commit apoptosis. Our study therefore aimed to find out the chronic effect of ketamine on neuron using prolonged incubation (48 h) of neuronal cells with ketamine in culture. Our results showed that differentiated neuronal cells were prone to the toxicity of ketamine but probably less susceptible than undifferentiated neuronal cells and f... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2757494 Mon, 31 Aug 2009 23:00:00 +0100 2757494 Differences between nanoscale structural and electrical properties of AZO:N and AZO used in polymer light-emitting diodes http://www.medworm.com/index.php?rid=2757493&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20775 Conducting atomic force microscopy and scanning surface potential microscopy were adopted to investigate the nanoscale surface electrical properties of N-doped aluminum zinc oxide (AZO:N) films that were prepared by pulsed laser deposition (PLD) at various substrate temperatures. Experimental results demonstrated that when the substrate temperature is 150°C and the N2O background pressure is 150 mTorr, the N-dopant concentration on the surface is optimal. In addition, the root-mean-square roughness value of the film surface, the low contact current ( (Source: Microscopy Research and Technique) Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2757493 Mon, 31 Aug 2009 23:00:00 +0100 2757493 Time-series investigation of fused vesicles in microvessel endothelial cells with atomic force microscopy http://www.medworm.com/index.php?rid=2757492&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20766 Vesicles or caveolae within endothelial cells, fusing together to form vacuolar organelles, are implicated in macromolecular transport and cellular element transmigration across the blood-brain barrier (BBB) during inflammation and ischemia. Vacuolar organelles have been described by transmission electron microscopy and immunofluorescence, but the details of their dynamics have not been well addressed yet. Herein, by using tapping mode atomic force microscopy (AFM), we observed the time-series changes of fused vesicles within the serum-free cultured rat cerebral microvessel endothelial cells. The fused vesicles were certainly proved by fluorescent staining of Fm4-64 combining simultaneous AFM imaging, as well as the field emission scanning electron microscopy technique. And energy dispersi... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2757492 Mon, 31 Aug 2009 23:00:00 +0100 2757492 A novel method for shape from focus in microscopy using Bezier surface approximation http://www.medworm.com/index.php?rid=2757491&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20765 In this article, we introduce a novel shape from focus method to compute 3D shape of microscopic objects, based on modified-pixel intensities and Bezier surface approximations. A new and simple but effective focus measure is proposed. In our focus measure, the original intensities of a sequence of small neighborhood are modified by subtracting the maximum of the values of first and last frames. An initial depth map is calculated by finding the maximum of the pixel's focused energy and its corresponding frame number. Missing information between two consecutive frames, false depth detection, and enhancement of noise related intensities may provide inaccurate depth map. To overcome these problems and to produce an accurate depth map, we proposed Bezier surface approximation. The proposed meth... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2757491 Mon, 31 Aug 2009 23:00:00 +0100 2757491 Automated compensation of light attenuation in confocal microscopy by exact histogram specification http://www.medworm.com/index.php?rid=2757490&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20767 Confocal laser scanning microscopy (CLSM) enables us to capture images representing optical sections on the volume of a specimen. The images acquired from different layers have a different contrast: the images obtained from the deeper layers of the specimen will have a lower contrast with respect to the images obtained from the topmost layers. The main reasons responsible for the effects described above are light absorption and scattering by the atoms and molecules contained in the volume through which the light passes. Also light attenuation can be caused by the inclination of the observed surface. In the case of the surfaces that have a steep inclination, the reflected light will have a different direction than the one of the detector. We propose a technique of digital image processing t... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2757490 Mon, 31 Aug 2009 23:00:00 +0100 2757490 MRT letter: Recovering weak-textured surfaces using image focus http://www.medworm.com/index.php?rid=2757489&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20768 This article presents a new SFF algorithm which can compute precise depth of dense as well as weak textured objects. Segmentation is applied to discard wrong depth estimate and then later interpolating them from accurate depth values of their neighbors. The performance of the proposed method is tested, using different image sequences of synthetic and real objects, with varying textures. Microsc. Res. Tech., 2009. © 2009 Wiley-Liss, Inc. (Source: Microscopy Research and Technique) Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2757489 Mon, 31 Aug 2009 23:00:00 +0100 2757489 Methodological assessment of acid-etching for visualizing the osteocyte lacunar-canalicular networks using scanning electron microscopy http://www.medworm.com/index.php?rid=2757488&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20772 Osteocytes are the most abundant of the bone cells. Each osteocyte is contained within its own lacuna and connected to adjacent osteocytes via fillipodial processes, which form an intricate network of canaliculi within the matrix. Studying this intricate network of cells and their processes is difficult, because it exists embedded within a densely mineralized matrix. Scanning electron microscopy (SEM) has been shown to be a useful tool for visualizing this cellular network, yet the techniques involved for preparing specimens has not been systematically explored. The goal of this study was to investigate how variations in acid-etching, both etching media and etching duration, affect SEM-based visualization of the osteocyte lacunar-canalicular network. Bone samples were embedded in plastic a... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2757488 Mon, 31 Aug 2009 23:00:00 +0100 2757488 The olfactory organ of the trout Salmo trutta fario: A novel localization for a progestin receptor http://www.medworm.com/index.php?rid=2757487&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20776 A progestin receptor (PR) has been detected in the olfactory organ of the trout Salmo trutta fario. The specificity of this receptor was high for 17[alpha],20[beta]-dihydroxy-4-pregnen-3-one (17[alpha],20[beta]-DP), but it also bound 17[alpha]-hydroxy-progesterone (17[alpha]-OHP) and 21-hydroxyprogesterone (21-OHP), even when present at low concentrations (10-fold in relative binding affinity assay). Progesterone (P) competed effectively at much higher concentrations (1,000-fold in relative binding affinity assay). Immunohistochemical studies carried out with three different monoclonal antibodies against human progesterone receptor (hPR), chicken progesterone receptor hinge region (cPR), and chicken progesterone receptor A/B domain (PR22), revealed that immunoreactivity was present in the ... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2757487 Mon, 31 Aug 2009 23:00:00 +0100 2757487 A model for predicting pathologist's velocity profiles when navigating virtual slides http://www.medworm.com/index.php?rid=2757486&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20760 This article presents a soft computing model, which permits to anticipate the pathologist trajectories in diagnosis tasks when exploring virtual slides. The Bayesian strategy combines an offline model of a baseline pathologist knowledge (the prior) and a prediction online module (the likelihood) that captures a particular pathologist navigation pattern. While optimal parameters for the biologically inspired offline model are calculated using an Expectation-Maximization strategy, prediction is carried out by a particle filter. Parameters are estimated from several series of actual navigations performed by several pathologists in different virtual slides. The present approach is compared with other conventional prediction methods and decreases the calculated MSE in about a 50% for the entire... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2757486 Mon, 31 Aug 2009 23:00:00 +0100 2757486 Cortactin/tyrosine-phosphorylated cortactin interaction with connexin 43 in mouse seminiferous tubules http://www.medworm.com/index.php?rid=2757485&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20771 Deletion of the cortactin gene leads to male infertility. Considering that cortactin is an actin filament (F-actin)-binding protein associated with intercellular junctions, we measured changes in the expression and distribution of cortactin and tyrosine phosphorylated cortactin (P-cortactin) in the seminiferous epithelium of developing and adult mice to address the physiological significance of cortactin to germ cell differentiation. Cortactin was expressed in neonatal and developing Sertoli cells. Cortactin levels decreased early during puberty, while P-cortactin increased. Cortactin labeling was intense in the basal and apical thirds of the epithelium. Sertoli cell cytoplasmic processes facing spermatogonia, preleptotene spermatocytes, and step 8-13 spermatids were intensely labeled by b... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2757485 Mon, 31 Aug 2009 23:00:00 +0100 2757485 Differentiation of Leydig cells in the Mongolian gerbil http://www.medworm.com/index.php?rid=2719539&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20763 In conclusion, the process of differentiation of ALC population in gerbil follows the pattern previously established for other rodents. However, the resulting mature ALC are strikingly different due their singular asymmetric morphology and presence of a cytoplasmic canaliculus and as well as their functional heterogeneity. Microsc. Res. Tech., 2009. © 2009 Wiley-Liss, Inc. (Source: Microscopy Research and Technique) Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2719539 Thu, 20 Aug 2009 23:00:00 +0100 2719539 An automatic integrated approach for stained neuron detection in studying neuron migration http://www.medworm.com/index.php?rid=2719540&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20762 Neurons that come to populate the six-layered cerebral cortex are born deep within the developing brain in the surface of the embryonic cerebral ventricles. It is very important to detect these neurons for studying histogenesis of the brain and abnormal migration that had been linked to cognitive deficits, mental retardation, and motor disorders. The visualization of labeled cells in brain sections was performed by immunocytochemical examination and its image data were documented to microscopic pictures. Based on the fact, automatic accurate neurons labeling is prerequisite instead of time-consuming manual labeling. In this article, a fully automated image processing approach is proposed to detect all the stained neurons in microscopic images. First of all, dark stained neurons are achieve... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2719540 Wed, 19 Aug 2009 23:00:00 +0100 2719540 Role of MMP9 on invadopodia formation in cells from adenoid cystic carcinoma. Study by laser scanning confocal microscopy http://www.medworm.com/index.php?rid=2674140&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20761 Migration, invasion and protease activity are essential for tumor progression and metastasis. Metastatic cells rely on invadopodia to degrade and invade extracellular matrix (ECM). Invadopodia are membrane protrusions with enzymes required for ECM degradation. These protrusions contain cortactin and membrane type 1 matrix metalloproteinase (MT1-MMP) superimposed to areas of digested matrix. Here we characterized invadopodia in a cell line (CAC2) derived from human adenoid cystic carcinoma. We carried out fluorescent-substrate degradation assay to assess in situ protease activity of CAC2 cells. Digestion spots in fluorescent substrate appear as black areas in green background. Cells were cultured on Matrigel-gelatin-FITC and fixed after 1 h and 3 h. CAC2 cells were double labeled to actin a... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2674140 Wed, 05 Aug 2009 23:00:00 +0100 2674140 Regenerating titanium ventricular assist device surfaces after gold/palladium coating for scanning electron microscopy http://www.medworm.com/index.php?rid=2653546&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20757 Titanium is one of the most commonly used materials for implantable devices in humans. Scanning electron microscopy (SEM) serves as an important tool for imaging titanium surfaces and analyzing cells and other organic matter adhering to titanium implants. However, high-vacuum SEM imaging of a nonconductive sample requires a conductive coating on the surface. A gold/palladium coating is commonly used and to date no method has been described to "clean" such gold/palladium covered surfaces for repeated experiments without etching the titanium itself. This constitutes a major problem with titanium-based implantable devices which are very expensive and thus in short supply. Our objective was to devise a protocol to regenerate titaniumsurfaces after SEM analysis. In a series of experiments, tita... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2653546 Wed, 29 Jul 2009 23:00:00 +0100 2653546 Is there a role for thyroid hormone on spermatogenesis? http://www.medworm.com/index.php?rid=2644959&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20759 Appropriate level of thyroid hormone is essential for normal development and metabolism in most vertebrate tissues and altered thyroid status impacts adversely on them. For many years the testis was regarded as a thyroid hormone unresponsive organ, but consistent evidence accumulated in the past two decades has definitively changed this classical view. Currently, the concept that thyroid hormone plays a critical role in testis development, in rats and other vertebrate species, is clearly established. Although the effects of thyroid hormone on Sertoli and Leydig cells in the immature testis are well described, its role on the adult organ remains controversial. In this review, we summarize and discuss the recent development on the thyroid hormone effects in immature and adult testes. Particu... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2644959 Mon, 27 Jul 2009 23:00:00 +0100 2644959 A novel and effective multistage classification system for microscopic starch grain images http://www.medworm.com/index.php?rid=2621414&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20758 This article presents a novel and effective multistage system for classifying Chinese Materia Medica microscopic starch grain images. The proposed classification system is constructed based on the Gaussian mixture model-based clustering, the feature assignment algorithm, and the similarity measurement. Several features for each starch grain image are extracted and every class of drug is represented by a set of characteristic features. For each stage of the system, only one feature is chosen and assigned to that stage via the feature assignment algorithm, and the corresponding characteristic features are subdivided into smaller subsets based on clustering techniques. At the final stage, each subset contains a certain class of drugs (with corresponding characteristic features) and similarity... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2621414 Mon, 20 Jul 2009 23:00:00 +0100 2621414 Morphological and crystallographic evolution of bainite transformation in Fe-0.15C binary alloy http://www.medworm.com/index.php?rid=2583996&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20755 In this article, an in situ observation method, combining laser scanning confocal microscopy and electron backscattering diffraction, was used to investigate the morphological and crystallographic evolution of bainite transformation in a Fe-0.15C binary alloy. The nucleation at a grain boundary and inclusions, sympathetic nucleation, and impingement event of bainitic ferrite were directly shown in real time. The variant evolution during bainite transformation and misorientation between bainitic ferrites were clarified. Strong variant selection was observed during sympathetic nucleation. Microsc. Res. Tech., 2009. © 2009 Wiley-Liss, Inc. (Source: Microscopy Research and Technique) Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2583996 Wed, 08 Jul 2009 23:00:00 +0100 2583996 New methods for chicken embryo manipulations http://www.medworm.com/index.php?rid=2576351&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20753 The capacity to image a growing embryo while simultaneously studying the developmental function of specific molecules provides invaluable information on embryogenesis. However, until recently, this approach was accomplished with difficulty both because of the advanced technology needed and because an easy method of minimizing damage to the embryo was unavailable. Here, we present a novel way of adapting the well-known EC culture of whole chick embryos to time-lapse imaging and to functional molecular studies using blocking agents. The novelty of our method stems from the ability to apply blocking agents ex ovo as well as in ovo. We were able to study the function of a set of molecules by culturing developing embryos ex ovo in tissue culture media containing these molecules or by injecting ... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2576351 Mon, 06 Jul 2009 23:00:00 +0100 2576351 Changes in MAPK pathway in neonatal and adult testis following fetal estrogen exposure and effects on rat testicular cells http://www.medworm.com/index.php?rid=2555970&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20756 Concerns have been raised about the possible role of the phytoestrogen genistein and the xenoestrogen Bisphenol A (BPA) as endocrine disruptors. In the present study, we examined the effects of fetal exposure to genistein and BPA on the Mitogen-activated protein kinase (MAPK) pathway and on testicular cell populations in neonatal and adult rat testes. At postnatal day (PND) 3, genistein (0.1-10 mg/kg/day) and BPA (1-200 mg/kg/day) induced Raf1 and Erk1/2 mRNA and protein increases in testes, mainly in Sertoli cells. No changes were seen for Mek1. At PND60, Erk1/2 protein expression remained robust in Sertoli cells and in some spermatogonia. Raf1 was predominant in Leydig cells while Mek1 was expressed strongly in spermatogonia, and they were both expressed in pachytene spermatocytes. No co... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2555970 Mon, 29 Jun 2009 23:00:00 +0100 2555970 A simple method for multiday imaging of slice cultures http://www.medworm.com/index.php?rid=2555971&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20750 We describe a simple method to image a slice culture preparation during the course of multiple days and over many continuous hours, without noticeable damage to the tissue or photobleaching. Our method uses a simple, inexpensive custom-built insulator constructed around the microscope to maintain controlled temperature and uses a perfusion chamber as used for in vitro slice recordings. Microsc. Res. Tech., 2009. © 2009 Wiley-Liss, Inc. (Source: Microscopy Research and Technique) Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2555971 Sun, 28 Jun 2009 23:00:00 +0100 2555971 Ontogeny of the androgen receptor expression in the fetal and postnatal testis: Its relevance on Sertoli cell maturation and the onset of adult spermatogenesis http://www.medworm.com/index.php?rid=2508276&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20754 From fetal life to adulthood, the testis evolves through maturational phases showing specific morphologic and functional features in its different compartments. The seminiferous cords contain Sertoli and germ cells, surrounded by peritubular cells, and the interstitial tissue contains Leydig cells and connective tissue. Sertoli cells secrete anti-Müllerian hormone (AMH), whereas Leydig cells secrete androgens. In the fetal and early postnatal testis, Leydig cells actively secrete androgens. Sertoli cells are morphologically and functionally immature - e.g., they secrete high levels of AMH - and germ cells proliferate by mitosis but do not enter meiosis. During infancy and childhood, Leydig cells regress and testosterone secretion declines dramatically. Sertoli cells remain immature and sp... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2508276 Wed, 24 Jun 2009 23:00:00 +0100 2508276 Structural study of the salivary glands of Anocentor nitens (Acari: Ixodidae) during the feeding cycle http://www.medworm.com/index.php?rid=2508278&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20751 The salivary glands of Anocentor nitens (Neumann,) occur in pairs and are located in the anterolateral region of the general cavity, with milky white color and approximately equal sizes. They consist of a secretory portion and an excretion duct. In some glandular acini, all the cells had a basophilic appearance they were stained by hematoxylin, whereas others presented cells with different staining affinities. In this work, we describe the variations observed in these glands during the feeding cycle of ticks [after feeding (0 h) and successively at 24, 48, 72, 96, 120, and 144 h]. The cells stained by hematoxylin were shown to be more reactive to Alcian blue, thus demonstrating the presence of acid glycosaminoglycans, whereas those stained using eosin presented weak or no reaction. A stron... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2508278 Thu, 18 Jun 2009 23:00:00 +0100 2508278 Self-assembled monolayers of DNA on cysteamine modified Au(111) surface: Atomic force microscopy study http://www.medworm.com/index.php?rid=2508277&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20752 The [lambda]-DNA molecules self-assemble on cysteamine-modified gold (111) surface to form flat-lying self-assembled monolayers (SAMs). The formation kinetics of such DNA SAMs is studied by atomic force microscopy (AFM). AFM results show that DNA molecules do not arrange themselves on cysteamine-modified gold (111) surface into a well-ordered monolayer. It is also found that the surface density of DNA monolayer does not increase as the DNA concentration increases. The high temperature of DNA solution and the immersing in ultrapure water produce some obvious DNA bundles. Whereas divalent cations in DNA solution result in the formation of more compact DNA films. The obtained information may be useful for practical application of the DNA films and further theoretical studies. Microsc. Res. Te... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2508277 Thu, 18 Jun 2009 23:00:00 +0100 2508277 PIV as a method for quantifying root cell growth and particle displacement in confocal images http://www.medworm.com/index.php?rid=2508279&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20749 In conclusion, PIV measured displacements successfully in images of living root cells and the external granular medium, revealing much potential for use by microscopists. Microsc. Res. Tech., 2009. © 2009 Wiley-Liss, Inc. (Source: Microscopy Research and Technique) Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2508279 Tue, 16 Jun 2009 23:00:00 +0100 2508279 Single photon fluorescent microlithography for live-cell imaging http://www.medworm.com/index.php?rid=2508285&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20748 Using fluorescent dyes to trigger the polymerization of a commercial polyurethane resin allows a rapid fabrication of micrometer and submicrometer sized fluorescent structures by one-photon absorption. Here, we show that standard He-Ne lasers emitting at 632.8 nm can be used to start the photopolymerization and that very low laser power is required. This procedure allows the fabrication of fiduciary fluorescent references on standard glass coverslips, mica sheets, or gold-coated coverslips for laser scanning or standard fluorescent microscopy. The biocompatibility of the polymerized resin with cells in culture was tested by growing Xenopus melanophores and a standard laser scanning microscope was used to demonstrate that it is possible to use equipment readily available in several laborato... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2508285 Thu, 11 Jun 2009 23:00:00 +0100 2508285 Authenticating and distinguishing the eight species of traditional Tibetan medicine "Meiduoluomi" by microscopic technique http://www.medworm.com/index.php?rid=2508284&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20725 Because of the morphological and macroscopic similarity, many species of Erigeron and Aster (Asteraceae) are confusable and usually used under the same name "Meiduoluomi" in traditional Tibetan medicine (TTM). To find an easy, quick, and reliable method to authenticate and distinguish the eight main medicinal plants of these species, the light microscope was used to reveal the morphoanatomic details. The fixed, sectioned, and stained plant materials and epidermis materials were studied by microscopic techniques. The results of the microscopic features are systematically described and illustrated, and comparison parameters are presented. Furthermore, a key to the eight species of "Meiduoluomi" was constructed. Microscopy can be unambiguously used to authenticate and distinguish the eight ma... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2508284 Thu, 11 Jun 2009 23:00:00 +0100 2508284 Regulated production of SnoN2 is a feature of testicular differentiation http://www.medworm.com/index.php?rid=2508283&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20739 The objective of this study was to determine the cellular localization of phosphorylated SMAD2/3 and the transcriptional repressor SnoN (Ski-like), a modifier of SMAD2/3 transcriptional activity, in mouse testes. Western blot established that only the smaller SnoN isoform, SnoN2, is produced in the testis. By immunohistochemistry, widespread phospho-SMAD2/3 distribution was observed in somatic and germ cells at all ages. In contrast, SnoN2 production was highly regulated, being detected only in gonocytes and interstitial cells at birth and in pachytene spermatocytes at puberty. In the adult, SnoN2 expression differed to that during the first wave, being ubiquitously expressed but exhibiting regulated nuclear localization. In another model of spermatogenic differentiation, the irradiated ra... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2508283 Thu, 11 Jun 2009 23:00:00 +0100 2508283 Effects of di(2-ethylhexyl) phthalate on gap and tight junction protein expression in the testis of prepubertal rats http://www.medworm.com/index.php?rid=2508282&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20741 The aim of this study was to analyze whether di(2-ethylhexyl) phthalate (DEHP), a Sertoli and Leydig cell toxicant, is able to induce alterations in the expression of testicular gap and tight junction proteins. DEHP was administered by gavage (1 g/5 mL corn oil/kg body weight/day) to 25-day-old male Sprague-Dawley rats for 2 days (DEHP-27d) and control rats were treated with corn-oil vehicle for 2 days (C-27d); animals were killed 24 h after the last treatment. Testes of DEHP-27d rats showed different degrees of germ cell sloughing of seminiferous tubules (ST). No alterations of the blood testis barrier (BTB) by lanthanum tracer study were observed. ST of DEHP-27d rats showed a milder immunofluorescence and more restricted expression of connexin-43 (Cx43) in the adluminal and basal compart... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2508282 Thu, 11 Jun 2009 23:00:00 +0100 2508282 Color-based tumor tissue segmentation for the automated estimation of oral cancer parameters http://www.medworm.com/index.php?rid=2508281&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20746 This article presents an automatic color-based feature extraction system for parameter estimation of oral cancer from optical microscopic images. The system first reduces image-to-image variations by means of color normalization. We then construct a database which consists of typical cancer images. The color parameters extracted from this database are then used in automated online sampling from oral cancer images. Principal component analysis is subsequently used to divide the color features into four tissue types. Each pixel in the cancer image is then classified into the corresponding tissue types based on the Mahalanobis distance. The aforementioned procedures are all fully automated; in particular, the automated sampling step greatly reduces the need for intensive labor in manual sampl... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2508281 Thu, 11 Jun 2009 23:00:00 +0100 2508281 Study of intussusceptive angiogenesis in inflammatory regional lymph nodes by scanning electron microscopy http://www.medworm.com/index.php?rid=2508280&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20747 The aim of the present study was to verify the occurrence of intussusceptive angiogenesis in blood vessels from submandibular lymph nodes responsible for lymphatic drainage of the tongue. A surgical wound inflicted on the ventral tongue of male Wistar rats and submandibular regional lymph nodes were evaluated at different postoperative periods. Scanning electron microscopy (SEM) was used to observe 123 lymph nodes at times 2, 3, 7, 10, 14, and 21 postoperative days. During the analysis of the vascular models with SEM, intussusceptive angiogenesis was observed in all groups evaluated. This was more extensive on the second and third postoperative days (83.33% and 80%, respectively), representing in these groups the expansion of the vascular chain of lymph nodes. At 21 postoperative days, int... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2508280 Thu, 11 Jun 2009 23:00:00 +0100 2508280 Measurement of edge verticality of optical recording bits on blu-ray discs using scanning probe microscopy http://www.medworm.com/index.php?rid=2446530&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20745 This work investigates recording bits on recordable and rewritable blu-ray discs using atomic force microscopy and conducting atomic force microscopy with high contrast, respectively. The geometric structure of the recording bits is clearly observed in images, which, when coupled with cross-section analysis, yields precise bit dimensions, and edge horizontal extended length values. The microscopic results are a valuable reference for increasing the recognition rate of digital signals in optical storage media. Furthermore, such a rapid and convenient measuring mode is an indispensable research tool for developing new recording materials and improving formation mechanisms. Microsc. Res. Tech., 2009. © 2009 Wiley-Liss, Inc. (Source: Microscopy Research and Technique) Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2446530 Sun, 31 May 2009 04:00:00 +0100 2446530 Expression of patched-1 and smoothened in testicular meiotic and post-meiotic cells http://www.medworm.com/index.php?rid=2446536&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20733 Desert hedgehog (Dhh) signaling plays an essential role in the normal development of the testis and in the process of spermatogenesis. Little is known about the involvement in spermatogenesis of the prototypic member of the family, Ptc1, which acts to suppress hedgehog signaling through Smoothened (Smo). Here, we have examined the expression of Ptc1, Smo, and Dhh in mouse and rat seminiferous epithelium. Our findings demonstrate that Ptc1 and Smo are expressed by primary spermatocytes and by round and condensing spermatids whereas Dhh is expressed by Sertoli cells. The findings suggest that Sertoli cells coordinate Dhh-dependent spermatogenesis events via Ptc1 and Smo prior to the first meiotic division and in postmeiotic (haploid) cells, particularly during the first half of spermiogenesi... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2446536 Fri, 29 May 2009 04:00:00 +0100 2446536 Apoptotic DNA fragmentation can be revealed in situ: An ultrastructural approach http://www.medworm.com/index.php?rid=2446535&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20735 A common pattern of apoptotic death is DNA cleavage, initially producing large fragments (50 kbp), followed by the production of nucleosomic/oligonucleosomic fragments. Nevertheless, apoptosis without DNA fragmentation, at least of the nucleosomic type, has been reported. To investigate the spatial relationship between DNA cleavage and chromatin condensation, we applied the TUNEL technique to the ultrastructural analysis of apoptotic cells. A modified method, utilizing a gold-conjugated antidigoxigenin antibody, was carried out on U937 versus Molt-4 cells, both exposed to UVB radiation or staurosporine treatment. Gold particle density in the different domains of apoptotic cells was evaluated by a four-way ANOVA test. Gold labelling was more strongly localised in condensed chromatin than in... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2446535 Fri, 29 May 2009 04:00:00 +0100 2446535 Substrate influence on cell shape and cell mechanics: HepG2 cells spread on positively charged surfaces http://www.medworm.com/index.php?rid=2446534&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20742 A human hepatoma cell line (HepG2) was cultured on positively and negatively charged polyelectrolytes. Cell/surface adhesion and cell shape evolution were followed with quartz microbalance with dissipation (QCM-D) and optical microscopy as a function of time, respectively. In particular, substrates coated with poly(ethyleneimine) (PEI) led to fast cell attachment and further spreading, with average maximum frequency [Delta]f = 79 Hz and dissipation [Delta]D = 40 × 10-6. On the contrary, no cell spreading was observed on poly(sodium-4-styrenesulfonate) (PSS), with [Delta]f = 33 Hz and [Delta]D = 4.5 × 10-6. Atomic force microscopy (AFM) was used to investigate the influence of cell shape on its mechanical properties. Considering the cells as an homogenous solid material, the corresponding... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2446534 Fri, 29 May 2009 04:00:00 +0100 2446534 A scanning electron microscopic study on the cephalic cuticular sensillae of different larval stages of Antheraea assamensis, an indigenous silkworm species of Northeast India http://www.medworm.com/index.php?rid=2446533&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20736 Scanning electron microscopy revealed the presence of a number of cuticular sensillae on the head of different larval stages of the "Muga" silk moth, Antheraea assamensis. These include four types of sensilla trichodea, bead-like folded sensilla, two types of sensilla basiconica, and sensory pegs. The study seems to be the first attempt in describing the morphology and distribution pattern of the cephalic cuticular sensillae of different larval stages of the silk moth Antheraea assamensis, an indigenous species of Northeast India. The possible significance of the sensillae in searching behavior of the larval stages during feeding is discussed with the help of available literature. Microsc. Res. Tech., 2009. © 2009 Wiley-Liss, Inc. (Source: Microscopy Research and Technique) Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2446533 Fri, 29 May 2009 04:00:00 +0100 2446533 Increased androgen receptor and remodeling in the prostatic stroma after the inhibition of 5-alpha reductase and aromatase in gerbil ventral prostate http://www.medworm.com/index.php?rid=2446532&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20740 Prostate require high levels of steroidogenic enzymes such as 5[alpha]-reductase (5[alpha]-r) and Aromatase (Aro) for the formation of active steroids. Dihydrotestosterone (DHT), the prostate dominant androgen, is converted from testosterone (T) by the action of 5[alpha]-r. Aro provides an alternative pathway for estrogen, via T aromatization. Since prostatic maintenance is dependent on both reciprocal stromal-epithelial interaction and regulation by steroids, this study aimed to elucidate what the absence of 5[alpha]-r and Aro enzymes provokes in the prostate microenvironment after their long-term inhibition. Data obtained 1 day after the 30 consecutive days of enzymatic inhibition with Finasteride (5[alpha]-r inhibitor) and Letrozole (Aro inhibitor) demonstrated a marked stromal remodeli... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2446532 Fri, 29 May 2009 04:00:00 +0100 2446532 Visualization of wound periderm and hyphal profiles in pine stems inoculated with the pitch canker fungus Fusarium circinatum http://www.medworm.com/index.php?rid=2446531&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20744 Postpenetration behavior of Fusariumcircinatum in stems of pine species was investigated with light and transmission electron microscopy. Two-year-old stems of Pinus rigida and P. densiflora were wound-inoculated with the fungal conidial suspension and subjected to 25°C for up to 30 days. It was common to observe the formation of wound periderm on each pine species, recovering wounded sites with newly formed tissues. The outermost thick layer of wound periderm was pink to red colored with the phloroglucinol-EtOH staining, indicating heavy deposition of lignin in wound periderm. The cork layers in the wound periderm of the two pine species consisted of cells that were mostly devoid of cellular contents in cytoplasm. The cork cells showed convoluted cell walls with different electron densit... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2446531 Fri, 29 May 2009 04:00:00 +0100 2446531 Distribution and three-dimensional appearance of the interstitial cells of Cajal in the rat stomach and duodenum http://www.medworm.com/index.php?rid=2423493&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20743 The relationship between the interstitial cells of Cajal (ICC) and enteric nerves or smooth muscles cells is not fully defined. Presently, distribution and appearance of ICC in the rat stomach and duodenum was studied by immunohistochemistry, electron microscopy, and three-dimensional reconstruction. c-kit expressing ICC were regularly observed in the Auerbach's myenteric plexus (AP) of the stomach and duodenum. ICC in stomach and duodenum muscle layers was dissimilarly distributed. c-kit immunoreactive cells were sparsely distributed in the stomach circular muscle layer but were abundant in the duodenum deep muscular plexus (DMP). Electron microscopy revealed that stomach ICC-AP were irregular ovals with few cytoplasmic processes, and possessed an electron-dense cytoplasm, numerous mitoch... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2423493 Wed, 20 May 2009 04:00:00 +0100 2423493 Imaging of cochlear tissue with a grating interferometer and hard X-rays http://www.medworm.com/index.php?rid=2423499&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20728 This article addresses an important current development in medical and biological imaging: the possibility of imaging soft tissue at resolutions in the micron range using hard X-rays. Challenging environments, including the cochlea, require the imaging of soft tissue structure surrounded by bone. We demonstrate that cochlear soft tissue structures can be imaged with hard X-ray phase contrast. Furthermore, we show that only a thin slice of the tissue is required to introduce a large phase shift. It is likely that the phase contrast image of the soft tissue structures is sufficient to image the structures even if surrounded by bone. For the present set of experiments, structures with low-absorption contrast have been visualized using in-line phase contrast imaging and a grating interferomete... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2423499 Tue, 19 May 2009 04:00:00 +0100 2423499 Impact of caspase activation in human spermatozoa http://www.medworm.com/index.php?rid=2423498&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20732 Caspases are central components in the apoptosis signaling cascade. The family of cysteine proteases transduces and enhances the apoptosis signal, and activation of effector caspases results in controlled cellular degradation. Although initially the presence of caspases in spermatozoa was controversially discussed in recent years, many studies demonstrated their activation in male germ cells. Activated apoptosis signaling results in decreased fertilizing capacity of the sperm. This review presents the current knowledge on the role of caspases in human sperm. Techniques of caspase monitoring are highlighted. With regard to the high impact of caspases on the sperm fertilizing potential, physiological and pathological settings of caspase activation and inactivation are discussed. Finally, the... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2423498 Tue, 19 May 2009 04:00:00 +0100 2423498 Application of microscopy in authentication and distinguishing of 11 Paris species in West Sichuan http://www.medworm.com/index.php?rid=2423497&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20726 In the trade and raw drug market, the medicinal plants of genus Paris are available in the form of rhizomes without any vouchers, thus making it difficult to identify and distinguish the different species of Genus Paris. Recent studies have shown that the species of Paris possess different chemical constituents, pharmacological activities, and efficiencies in clinical application. To distinguish 11 species of Paris collected from the western Sichuan province of China and ensure their safety and efficacy, in the present work, the microscopic characteristics of rhizomes and the crude drug powder of the 11 species of Paris were compared using a light microscope according to the usual microscopic techniques. The results of the microscopic features were systematically described and illustrated.... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2423497 Tue, 19 May 2009 04:00:00 +0100 2423497 Three-dimensional analysis of connexin43 gap junction in the ex vivo rat seminiferous tubules: Short-term effects of hormonal effectors http://www.medworm.com/index.php?rid=2423496&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20731 Cx43 gap junctions are essential for proliferation, differentiation, and apoptosis of germ cells during spermatogenesis. However, only few and indirect observations have been reported on the distribution of Cx43, the predominant Cx within the seminiferous tubules. In thepresent study, we developed an innovative method that allows visualization of the three- dimensional localization of Cx43 associated with gap junctions and their functionality in isolated spermatogenic stage-specific seminiferous tubules. Cx43 gap junctions were present between myoid cells, between Sertoli cells, and between Sertoli and germ cells. Cx43 levels and coupling were stage-dependent with higher values at stages VI-VIII of spermatogenesis and markedly reduced at stages IX-X. Short-term exposure of seminiferous tub... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2423496 Tue, 19 May 2009 04:00:00 +0100 2423496 Effect of two medium chain triglycerides-supplemented diets on synaptic morphology in the cerebellar cortex of late-adult rats http://www.medworm.com/index.php?rid=2423495&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20737 Ketogenic diets (KDs) have shown beneficial effects in experimental models of neurodegeneration, designating aged individuals as possible recipients. However, few studies have investigated their consequences on aging brain. Here, late-adult rats (19 months of age) were fed for 8 weeks with two medium chain triglycerides-supplemented diets (MCT-SDs) and the average area (S), numeric density (Nvs), and surface density (Sv) of synapses, as well as the average volume (V), numeric density (Nvm), and volume density (Vv) of synaptic mitochondria were evaluated in granule cell layer of the cerebellar cortex (GCL-CCx) by computer-assisted morphometric methods. MCT content was 10 or 20%. About 10%MCT-SD induced the early appearance of senescent patterns (decreased Nvs and Nvm; increased V), whereas ... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2423495 Tue, 19 May 2009 04:00:00 +0100 2423495 Oleic acid-added embedding medium for histological analysis of hard tissue http://www.medworm.com/index.php?rid=2423494&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20738 In this study, mixtures of methyl methacrylate (MMA), di-butyl-phthalate (DBP), and oleic acid (OA) were tested for possible application as an embedding medium for large and small undecalcified bone specimens. Mechanical properties were tested in a compressive mode. We investigated the change of hydrophilicity in the sectioned surface by measuring the contact angle depending on the OA. Crystallinity was analyzed using a X-ray diffractometer (XRD). Surface analysis was performed using a confocal laser scanning microscope. To determine the staining efficiency of staining dyes, hamatoxylin-eosin (H&E) and Masson's trichrome (MT) staining methods were performed for the histological analysis of bone-implant complex. We confirmed that the investigated embedding media showed good properties such ... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2423494 Tue, 19 May 2009 04:00:00 +0100 2423494 Gene expression analysis of immunostained endothelial cells isolated from formaldehyde-fixated paraffin embedded tumors using laser capture microdissection - A technical report http://www.medworm.com/index.php?rid=2397718&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20734 This report unveils a method for quantitative gene expression analysis in cells that were identified by immunostaining and retrieved by LCM from FFPE tissues. This method is ideally suited for the analysis of relatively rare cell types within a tissue, and should improve on our ability to perform differential diagnosis of pathologies as compared to conventional LCM. Microsc. Res. Tech., 2009. © 2009 Wiley-Liss, Inc. (Source: Microscopy Research and Technique) Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2397718 Fri, 08 May 2009 04:00:00 +0100 2397718 Effect of black tea on histological and immunohistochemical changes in pancreatic tissues of normal and streptozotocin-induced diabetic mice (Mus musculus) http://www.medworm.com/index.php?rid=2397720&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20721 The aim of the present study is to evaluate the effect of hot water extract of black tea in regenerating [beta] cells in streptozotocin-induced diabetic mice. Light microscopic examination of pancreatic sections of streptozotocin-induced diabetic mice showed the acinar cells to be small, shrunken, and with deteriorated [beta] cells. The dose of streptozotocin not only altered the function of [beta] cells but also damaged the acinar region. The changes in acinar cells were coarsening of endoplasmic reticulation suggesting alteration in their secretory function. The control pancreatic tissue showed well-defined granulated islets and dark [beta] cells when stained with chrome hematoxylin and phloxine. Interestingly, pancreatic sections of diabetic mice fed with black-tea extract showed regene... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2397720 Thu, 07 May 2009 04:00:00 +0100 2397720 Microscopic studies of the influence of main exposure time on parameters of flexographic printing plate produced by digital thermal method http://www.medworm.com/index.php?rid=2397721&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20719 The digital thermal technology of producing flexographic printing plates from photopolymer plates is one of the newest technologies. This technology allows to develop flexographic plates without the use of any solvent. The process of producing flexographic printing plates by the digital thermal method consists of several main stages: back exposure, laser exposure, main exposure, thermal development, post exposure, and light finishing. The studies carried out with the use of optical stereoscopic microscopy allowed to determine the effect of time of main exposure to ultraviolet radiation on the dot area, diameter, and edge factor of halftone dots reproduced on flexographic printing plate produced by the digital thermal method, as well as on the quality of reproducing the surface and on the p... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2397721 Wed, 06 May 2009 04:00:00 +0100 2397721 [gamma]H2AX marks the main events of the spermatogenic process http://www.medworm.com/index.php?rid=2380570&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20730 Recent findings reveal that chromatin reorganization requires formation and repair of DNA double strand breaks (DSBs). Therefore, dynamic changes in chromatin structure taking place along the mitotic and the meiotic processes should be accompanied by histone H2AX phosphorylation and formation of [gamma]H2AX, a prominent event occurring at DSB sites. Accordingly, [gamma]H2AX has been detected at multiple developmental steps in adult germ cells. Nevertheless, no comprehensive study focusing on the main biological events occurring in the seminiferous epithelium has been carried out in mammals. Here I analyze H2AX phosphorylation in immunostained testis sections comparing PAS/cresyl violet counterstained, noncounterstained, and immuno-fluorescence preparations and show several waves of H2AX ph... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2380570 Thu, 30 Apr 2009 04:00:00 +0100 2380570 Strategic aspects of spermatogenesis and the maturation of spermatozoa: Introduction http://www.medworm.com/index.php?rid=2375445&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20729 No abstract. (Source: Microscopy Research and Technique) Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2375445 Wed, 29 Apr 2009 04:00:00 +0100 2375445 Ultrastructural analysis of radicular dentine surface submitted to CO2 laser at different parameters http://www.medworm.com/index.php?rid=2341651&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20723 Conclusions: The intensity of the effects is dependent on the laser-irradiation dosimetries. Alterations were more intense when higher parameters were used. Microsc. Res. Tech., 2009. © 2009 Wiley-Liss, Inc. (Source: Microscopy Research and Technique) Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2341651 Sat, 18 Apr 2009 04:00:00 +0100 2341651 Expression profile of a 400-bp Stra8 promoter region during spermatogenesis http://www.medworm.com/index.php?rid=2341654&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20724 In this study, a detailed analysis of the EGFP expression pattern in the testis was carried out, showing that the transgene was expressed in meiotic and postmeiotic germ cells and not in undifferentiated germ cells. These findings were supported by confocal microscopy and flow cytometric analyses, and do not agree with the previous report concerning the 400-bp Stra8 promoter activity. Microsc. Res. Tech. 2009. © 2009 Wiley-Liss, Inc. (Source: Microscopy Research and Technique) Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2341654 Fri, 17 Apr 2009 04:00:00 +0100 2341654 Integrated analytical study for the authentication of five Russian icons (XVI-XVII centuries) http://www.medworm.com/index.php?rid=2341653&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20727 This article presents an integrated approach for the authentication of five Russian icons through the study of their paint materials (elemental composition and stratigraphic mapping), state of conservation and artistic technique. The results of the applied analytical protocol, involving optical and scanning electron microscopy coupled with energy-dispersive X-ray spectrometry and Fourier transformed infrared spectroscopy/microspectroscopy, led to the conclusion that the five icons belong to the same group and iconographic school (from Northern Russia) and date from the end of the 16th century to the beginning of 17th century. Because of massive overpainting, the icons "suffered" a change in iconographic style and significance, and restoration treatment brought to light the original paintin... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2341653 Fri, 17 Apr 2009 04:00:00 +0100 2341653 Micro-fabrication and monitoring of three-dimensional microstructures based on laser-induced thermoplastic formation http://www.medworm.com/index.php?rid=2341652&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20722 This article reports a novel laser-induced micro-fabrication method and its monitoring system for three-dimensional (3D) microstructures. The mechanism of the method is that a small zone of thermoplastic material melted by laser heating grows in liquid surrounding environment, solidifying into a convex microstructure, such as micro-dot or micro-pillar. A laser diode (808 nm) with maximum power output of 130 mW is used as power source, and a kind of paraffin mixed with stearic acid and paint serves as the thermoplastic material for 3D microstructure formation experiments. A light microscope system consisting of a charge-coupled device (CCD) and a computer is utilized to realize real-time observation of the micro-fabricating process. The distribution of local temperature rise on material sur... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2341652 Fri, 17 Apr 2009 04:00:00 +0100 2341652 Simultaneous 3D visualization and quantification of murine bone and bone vasculature using micro-computed tomography and vascular replica http://www.medworm.com/index.php?rid=2330120&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20720 Recent evidence suggests a close functional relationship between osteogenesis and angiogenesis as well as between bone remodeling and bone vascularization. Consequently, there is a need for visual inspection and quantitative analysis of the bone vasculature. We therefore adapted and implemented two different vascular corrosion casting (VCC) protocols using a polyurethane-based casting resin in mice for a true three-dimensional (3D), direct, and simultaneous measurement of bone tissue and vascular morphology by micro-computed tomography ([mu]CT). For assessment of vascular replicas at the level of capillaries, a vascular contrast perfusion (VCP) protocol was devised using a contrast modality based on a barium sulfate suspension in conjunction with synchrotron radiation (SR) [mu]CT. The vasc... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2330120 Fri, 17 Apr 2009 04:00:00 +0100 2330120 Measuring microenvironment mechanical stress of rat liver during diethylnitrosamine induced hepatocarcinogenesis by atomic force microscope http://www.medworm.com/index.php?rid=2330123&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20716 We developed a highly sensitive method to detect liver tissue stiffness with atomic force microscopy (AFM), and investigated the physical features of hepatocarcinogenesis. Wistar rats received weekly intraperitoneal injections of diethylnitrosamine (DEN) or saline (control) followed by a 2-week wash-out period. Liver samples were harvested at 10, 14, or 18 weeks for pathological examination and stress detection. Previously normal liver tissues developed fibrosis and carcinoma after DEN administration. Although the elastic modulus (E) values of the normal (saline; 0.18 ± 0.04 MPa), fibrotic (8 weeks DEN; 0.25 ± 0.06 MPa) and cirrhotic (12 weeks DEN; 0.39 ± 0.06 MPa) tissues were significantly different, there was no significant difference between the E values of the cirrhotic and the hep... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2330123 Tue, 07 Apr 2009 04:00:00 +0100 2330123 5-HT 1A and 2A receptor positive cells in the cerebella of mice and human and their decline during aging http://www.medworm.com/index.php?rid=2330122&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20717 This study evaluated the 5-HT1A and 5-HT2A receptor positive cells in the cerebella of mice and human by immunocytochemistry. Mice were of ages 1, 3, and 12 months whereas the human subjects were divided into two groups, a younger 57-78 years old group and an older 82-91 years old group. Both 5-HT1A and 5-HT2A receptor positive cells were observed in the molecular and granular layers of the cerebella of mice and human. Although there was a decline in these positive cells during aging, no regional difference in the positive cells were observed in the anterior, middle, and posterior regions of the cerebella. Microsc. Res. Tech. 2009. © 2009 Wiley-Liss, Inc. (Source: Microscopy Research and Technique) Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2330122 Tue, 07 Apr 2009 04:00:00 +0100 2330122 Ultrastructural changes of platelet aggregates and fibrin networks in a patient with renal clear cell adenocarcinoma: A scanning electron microscopy study http://www.medworm.com/index.php?rid=2330121&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20718 A 47-year-old male patient presented with weight loss, hematuria, and a left renal mass, which proved to be a clear cell renal carcinoma with multiple liver, pulmonary and bone metastases. The platelet count was raised initially (414 × 109/L) but declined 10 weeks after a debulking procedure followed by chemotherapy. Fibrin clots were prepared for scanning electron microscopy (SEM) by adding human thrombin to platelet rich plasma (derived by differential centrifugation of fresh blood samples taken from the patient as well as controls). The clots were washed, fixed in 2.5% glutaraldehyde and Dulbecos phosphate buffered saline and prepared for SEM with a Zeiss Ultra 55 FEG SEM. The SEM photographs revealed an altered morphology of the platelet aggregates with multiple breakages in the plate... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2330121 Tue, 07 Apr 2009 04:00:00 +0100 2330121 Morphological filter improve the efficiency of automated tracking of secretory vesicles with various dynamic properties http://www.medworm.com/index.php?rid=2330124&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20711 Membrane trafficking is a very important physiological process involved in protein transport, endocytosis, and exocytosis. The functions of vesicles are strongly correlated with various spatial dynamic properties of vesicles, including their types of movements and morphology. Several methods are used to quantify such dynamic properties, but most of them are specific to particular populations of vesicles. We previously developed the so-called PTrack system for quantifying the dynamics of secretory vesicles near the cell surface, which are small and move slowly. To improve the system performance in quantifying large and fast-moving vesicles, we firstly combined morphological filter with two-threshold image processing techniques to locate granules of various sizes. Next, Kalman filtering was ... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2330124 Mon, 06 Apr 2009 04:00:00 +0100 2330124 Morphometric characterization of murine articular cartilage - Novel application of confocal laser scanning microscopy http://www.medworm.com/index.php?rid=2330126&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20713 A new technique for characterization of the three-dimensional morphology of murine articular cartilage is proposed. The technique consists of a novel application of confocal laser scanning microscopy (CLSM), where the objective was to develop and validate it for cartilage measurements in murine joints. Murine models are used in arthritis research, because they are well-described for manipulating the disease pathophysiology, facilitating our understanding of the disease, and identifying new targets for therapy. A calibration and reproducibility study was carried out to provide a consistent testing methodology for quantification of murine joints. The proximal tibial condyles from male C57BL/6 mice were scanned using a CLS microscope with an isotropic voxel size of 5.8 [mu]m. Measurements and... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2330126 Thu, 02 Apr 2009 04:00:00 +0100 2330126 Ultrastructure and distribution of superficial neuromasts of blind cavefish, Phreatichthys andruzzii, juveniles http://www.medworm.com/index.php?rid=2330125&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20714 Transmission and scanning electron microscopy (TEM, SEM) were used to study the ultrastructure of superficial neuromasts in 15 six-month old blind cavefish juveniles, Phreatichthys andruzzii (Cyprinidae). In five specimens examined with SEM, the number of superficial neuromasts over the fish body (480-538) was recorded. They were localized mainly on the head (362-410), including the dorsal surface, the mentomandibular region, and laterally from the mouth to the posterior edge of the operculum. Neuromasts were also present laterally on the trunk and near the caudal fin (116-140). A significantly higher number of neuromasts were present on the head compared to the trunk (t-test, P < 0.05). Superficial neuromasts of the head and those along the trunk were similar in ultrastructure. Each neuro... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2330125 Thu, 02 Apr 2009 04:00:00 +0100 2330125 How to visualize the spider mite silk? http://www.medworm.com/index.php?rid=2300731&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20712 Tetranychus urticae (Acari: Tetranychidae) is a phytophagous mite that forms colonies of several thousand individuals. Like spiders, every individual produces abundant silk strands and is able to construct a common web for the entire colony. Despite the importance of this silk for the biology of this worldwide species, only one previous study suggested how to visualize it. To analyze the web structuration, we developed a simple technique to dye T. urticae'silk on both inert and living substrates. Fluorescent brightener 28 (FB) (Sigma F3543) diluted in different solvents at different concentrations regarding the substrate was used to observe single strands of silk. On glass lenses, a 0.5% dimethyl sulfoxide solution was used and on bean leaves, a 0.1% aqueous solution. A difference of silk ... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2300731 Thu, 02 Apr 2009 04:00:00 +0100 2300731 Distribution of ghrelin peptide in the gastrointestinal tract of stomachless and stomach-containing teleosts http://www.medworm.com/index.php?rid=2300733&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20709 The occurrence and localization of ghrelin peptide in the gastrointestinal tract of Carassius auratus and Dicentrarchus labrax, two fish species that exhibit different feeding behavior, different habitats, and different anatomical organizations of the gastroenteric tract, were examined by immunohistochemical methods and western blotting analysis. All of the gastrointestinal segments studied displayed immunohistochemical localizations of ghrelin peptide. Numerous single or clustered immunoreactive cells were found along the gastric folds, particularly in the pyloric region of Dicentrarcus labrax, whereas scattered ghrelin immunoreactive cells were observed in the intestinal epithelium of both fish species. Double immunolabeling PGP 9.5/ghrelin demonstrated the localization of ghrelin peptid... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2300733 Wed, 25 Mar 2009 04:00:00 +0100 2300733 Ultrastructural study of the ability of seminal plasma proteins to protect ram spermatozoa against cold-shock http://www.medworm.com/index.php?rid=2300732&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20710 In this study, we have attempted to examine the potential protective effect of SP proteins in membrane ultrastructure of ram spermatozoa subjected to cold-shock, by means of transmission electron microscopy (TEM). All the experiments were carried out with fresh spermatozoa freed from SP by a dextran/swim-up procedure. The high proportion of viable spermatozoa found in the swim-up obtained sample decreased drastically after the cold-shock treatment, and a considerable blebbing and vesiculation of the plasma and acrosomal membranes was found. The addition of SP proteins increased the sperm resistance to damage due to cold-shock (48% membrane-intact spermatozoa versus 15% in the control sample), and TEM analysis revealed that membrane alteration was prevented. This protective effect seems to ... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2300732 Wed, 25 Mar 2009 04:00:00 +0100 2300732 The dynamic epigenetic program in male germ cells: Its role in spermatogenesis, testis cancer, and its response to the environment http://www.medworm.com/index.php?rid=2300734&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20715 Spermatogenesis is a truly remarkable process that requires exquisite control and synchronization of germ cell development. It is prone to frequent error, as paternal infertility contributes to 30-50% of all infertility cases; yet, in many cases, the mechanisms underlying its causes are unknown. Strikingly, aberrant epigenetic profiles, in the form of anomalous DNA and histone modifications, are characteristic of cancerous testis cells. Germ cell development is a critical period during which epigenetic patterns are established and maintained. The progression from diploid spermatogonia to haploid spermatozoa involves stage- and testis-specific gene expression, mitotic and meiotic division, and the histone-protamine transition. All are postulated to engender unique epigenetic controls. In su... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2300734 Tue, 24 Mar 2009 04:00:00 +0100 2300734 Different expression of NR2B and PSD-95 in rat hippocampal subregions during postnatal development http://www.medworm.com/index.php?rid=2285250&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20708 In this study, we used immunofluorescent staining to assess NR2B and PSD-95 expressions and the relationship between them in CA1, CA3, and DG of rat hippocampus on different postnatal day. In CA1, NR2B expression decreased with age. It was high at birth, reached a plateau at P4, and declined gradually after P7. In CA3, NR2B expression was similar to that in CA1. But the stratum lucidum was devoid of staining. In DG, the NR2B expression retained a higher level. From P0 to P2, the PSD-95 expression in CA1 increased gently, and then declined slightly. After P7, the PSD-95 expression increased sharply till P28, and decreased again. In CA3 and DG, the PSD-95 expression is very similar except that low-level of PSD-95 was found in the CA3 stratum lucidum. The expression of NR2B did not correlate ... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2285250 Tue, 24 Mar 2009 04:00:00 +0100 2285250 An immunofluorescence method for postembedded tissue in the acrylic resin Technovit 9100 New® using fluorescein isothiocyanate secondary detection http://www.medworm.com/index.php?rid=2274990&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20705 This article describes a method for immunofluorescence labeling in Technovit 9100 New resin embedded tissue and suggests the likely chromogenic elements generating autofluorescence. Microsc. Res. Tech., 2009. © 2009 Wiley-Liss, Inc. (Source: Microscopy Research and Technique) Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2274990 Thu, 19 Mar 2009 04:00:00 +0100 2274990 Redundancy of the effect of TGF[beta]1 and [beta]-NGF on the second meiotic division of rat spermatocytes http://www.medworm.com/index.php?rid=2274992&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20706 In this study, we raised the question if [beta]-NGF and TGF[beta]1 have additional or redundant effects on this step. Hence, we addressed the effect of [beta]-NGF in combination with TGF[beta]1, as compared to those of [beta]-NGF or TGF[beta]1 separately, on the completion of meiosis by rat late PS/DS. Identification and counting of meiotic cells were performed by cytological methods. Under our culture conditions, some late PS/DS differentiated into round spermatids. When [beta]-NGF in combination with TGF[beta]1 was added to the culture medium, the numbers of PS/DS and of secondary spermatocytes were not modified by the treatment. By contrast, the number of round spermatids was about 2-fold lower in treated cultures than in basal cultures, and an increase in metaphase II, but not metaphas... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2274992 Mon, 16 Mar 2009 04:00:00 +0100 2274992 Comparative ultrastructure of nonwounded Mexican lime and Yuzu leaves infected with the citrus canker bacterium Xanthomonas citri pv. citri http://www.medworm.com/index.php?rid=2274991&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20707 Ultrastructural aspects of citrus canker development were investigated in nonwounded leaves of citrus species by transmission electron microscopy (TEM). A susceptible species Mexican lime and a resistant species Yuzu were spray-inoculated with a virulent strain of Xanthomonas citri pv. citri. Initial symptoms occurred on Mexican lime [sim]9 days after inoculation, whereas they appeared on Yuzu mostly 11 days after inoculation. In Mexican lime leaves, the bacterial invasion was usually accompanied by host cell wall dissolution and cellular disruption. Fibrillar materials from degenerated cell walls were usually found in intercellular spaces. Damaged host cells with necrotic cytoplasm showed the localized separation of plasma membrane from the cell wall. Bacterial multiplication and electron... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2274991 Mon, 16 Mar 2009 04:00:00 +0100 2274991 Spermatozoal RNAs: What about their functions? http://www.medworm.com/index.php?rid=2261056&cid=s_33602_166_f&fid=33602&url=http%3A%2F%2Fdx.doi.org%2F10.1002%252Fjemt.20697 The profound architectural changes that transform spermatids into spermatozoa result in a high degree of DNA packaging within the sperm head. However, the mature sperm chromatin that harbors imprinted genes exhibits a dual nucleoprotamine/nucleohistone structure with DNase-sensitive regions, which could be implicated in the establishment of efficient epigenetic information in the developing embryo. Despite its apparent transcriptionally inert state, the sperm nucleus contains diverse RNA populations, mRNAs, antisense and miRNAs, that have been transcribed throughout spermatogenesis. There is also an endogenous reverse transcriptase that may be activated under certain circumstances. It is now commonly accepted that sperm can deliver some RNAs to the ovocyte at fertilization. This review pre... Microscopy Research and Technique journals http://www.medworm.com/rss/comments.php?id=2261056 Sat, 14 Mar 2009 04:00:00 +0100 2261056