bluuebunny/crossref_metadata_2025_split huggingface dataset:
SEM and fluorescence imaging of callose deposition in root hair tips and suspension cultures of Streptanthus tortuosus
Authors: Walters K.S. , Sjolund R.D. , Moore K.C. 
Published: June 2020

Callose, B-1,3-glucan, a component of cell walls, is associated with phloem sieve plates, plasmodesmata, and other cell wall structures that are formed in response to wounding or infection. Callose reacts with aniline blue to form a fluorescent complex that can be recognized in the light microscope with ultraviolet illumination. We have identified callose in cell wall protuberances that are formed spontaneously in suspension-cultured cells of S. tortuosus and in the tips of root hairs formed in sterile callus cultures of S. tortuosus. Callose deposits in root hairs are restricted to root hair tips which appear to be damaged or deformed, while normal root hair tips lack callose deposits. The callose deposits found in suspension culture cells are restricted to regions where unusual outgrowths or protuberances are formed on the cell surfaces, specifically regions that are the sites of new cell wall formation.Callose formation has been shown to be regulated by intracellular calcium levels.
Electron Energy Loss Spectroscopy as an alternative method to follow up the route of activators and drugs in cells and tissues
Authors: Wolf B. , Uhl B. , Schwinde A. , Hauschildt S. , Bessler W.G. 
Published: June 2020

Investigations on uptake and distribution of small molecules and drugs in cells and cells from solide tissues are very important for the evaluation of their physiological behaviour and/or therapeutical potency. In electron microscopy localizations of molecules are usually performed by immunocytochemical techniques using antibodies coupled to electron dense markers (e.g. ferritin, gold). Problems concerning labelling precision and the production of highly specific antibodies to small molecules resulted in the evaluation of a more sensitive and more physical technique.We applied analytical electron energy loss spectroscopy (EELS) on ultrathin cryosections of different cells incubated with various substances (lipopeptide, 5’fluoro-uracil, cis-platinum, etc.) to follow up the route of these molecules from the plasma membrane to ultrastructural target regions. Using cryosections, investigations on almost native material, without the use of disintegrative methods, are possible. The main advantage of this procedure is the exclusion of metabolic interferences after the incubation period.Here, we demonstrate analytical energy loss spectroscopy as a possibility to analyze the distribution of small molecules within cells directly without the use of labelled antibodies.
Study on erythrocytes by SEM photogrammetry (SEMP) technique
Authors: Wensheng Tong , Lianhuang Lu , Zhijun Zhang 
Published: June 2020

This is a combined study of two diffirent branches, photogrammetry and morphology of blood cells. The three dimensional quantitative analysis of erythrocytes using SEMP technique, electron computation technique and photogrammetry theory has made it possible to push the study of mophology of blood cells from LM, TEM, SEM to a higher stage, that of SEM P. A new path has been broken for deeply study of morphology of blood cells.In medical view, the abnormality of the quality and quantity of erythrocytes is one of the important changes of blood disease. It shows the abnormal blood—making function of the human body. Therefore, the study of the change of shape on erythrocytes is the indispensable and important basis of reference in the clinical diagnosis and research of blood disease.The erythrocytes of one normal person, three PNH Patients and one AA patient were used in this experiment. This research determines the following items: Height;Length of two axes (long and short), ratio; Crevice in depth and width of cell membrane; Circumference of erythrocytes; Isoline map of erythrocytes; Section map of erythrocytes.
Ultra grids for observation of same wide specimen at low and high magnification
Authors: Hirose Haruko , Shirota Kohei , Makita Takashi 
Published: June 2020

Inconvenience in observation of wide view of biological sections was to prepare two sections from same sample; one thicker and heavily stained for contrasity at lower magnification and another ultrathin and lightly stained for higher magnifications. High contrasity and less deformative characteristics of currently developed electron microscopes,EM-002A(120kv) and EM-002B(200kv) from ISI.ABT,allow us to examine same section at both low and high magnifications.Besides to superwide(Fig.1-a) and wide (Fig.1-b) grid of 7mmØ those which have been utilized for super wide field electron microscopy using a specially constructed conventional electron microscope(LEM-2000, ISI,ABT)(1), a new type of ultra grid was designed in this laboratory(Fig. 1-c) and it became commercially available.Though wide- and superwide- grids have wider squares than those in ordinary 3mmØ grids, we have to cut off some bars with a razor blade to obtain further wider vision in some cases.(2-6). Ultra grid(Fig.1-c,Fig.2) eliminated that inconvenience since it has several types of large compartments.The largest square is 1.5×2.Omm and equivalent to 9 squares in super wide grid.
En bloc staining available for stereoscopic observation of epoxy resin Quetol 651-embedded thick sections under a 300 kV TEM
Authors: Kushida Tsuyuka , Iijima Haruyuki , Kushida Hiroshi , Tsuruta Chusei 
Published: June 2020

A staining method has been devised for easy en bloc staining for stereoscopic observation of epoxy resin Quetol 651-embedded thick sections under a 300 kV transmission microscope (TEM). In order to enhance staining properties in thick section, osmium tetroxide-fixed tissue blocks are stained only en bloc, since the images of both sides in thick section give high contrast and the image of an intermediate layer shows low contrast by double staining.This method uses carbohydrazide (Polysciences, Inc., U.S.A.) as osmium bridging agent, and both osmium tetroxide and uranyl acetate as electron staining agents.The following procedure is suitable for en bloc staining.
1.Fix small tissue blocks in 2% cacodylate-buffered osmium tetroxide (pH 7.4) for 3 hours at 4°C.2.Wash well in buffer for 1 hour.3.Transfer in 1% aqueous carbohydrazide for 2 hours at room temperature.4.Wash well in distilled water for 1 hour.5.Stain in 1% aqueous osmium tetroxide for 2 hours at room temperature.6.Wash well in distilled water for 1 hour.7.Dehydrate in 50% alcohol for 1 hour.8.Stain in a 2.5% solution of uranyl acetate in 50% alcohol for 3 hours at room temperature.9.Wash in 50% alcohol for 1 hour.10.Dehydrate with 60%, 70%, 80%, 90% and 100% (2 changes) alcohols for 30 minutes each.11.Embed in a mixture of Quetol 651 (Nissin EM Co., Ltd., Japan), nonenyl succinic anhydride, methyl nadic anhydride and DMP-30 according to the method of Kushida et al.
Localization of α1-proteinase inhibitor binding to extracellular matrix by tandem scanning reflected light and transmission electron microscopy
Authors: Roemer E.J. , Rinehart A.R. , Simon S.R. 
Published: June 2020

α1-Proteinase Inibitor (α1-PI), a major inhibitor of human leukocyte elastase (HLE), has been shown to bind saturably to extracellular matrix (ECM), composed of 51% glycoproteins and proteoglycans, 37% types I and III collagen and 12% elastin, produced by R22 rat aortic smooth muscle cells. To determine the components of the ECM to which α1-PI binds we have employed both tandem scanning reflected light (TSM) and transmission electron (TEM) microscopy on whole mount and sectioned material immunolabeled with antibodies to α1-PI and to various ECM components.R22 cells were grown on glass coverslips for TSM, on coverslips with formvar and carbon coated gold grids for whole mount TEM, and in 60 mm dishes for embedment. At confluence ascorbic acid was added for 6-18 days. The cells were lysed with 5 mM NH4OH at 25°C and the remaining ECM was rinsed with PBS and dH2O. ECM was blocked with 0.1% BSA in PBS for 30 min at 25°C and then incubated with 1.3 mg α1-PI/ml in 0.1% BSA/PBS for 1.5-2 h at 37°C.
Effects of ultrasound treatment of the arteria thoracica (A. mammaria) interna (ATI) during preparation for coronary artery bypass surgery (CABG): A correlative light microscopic, scanning and Transmission electron microscopy study
Authors: Muss Wolfgang , Lametschwandtner A. , Schistek R. , Ghosh P.K. , Thurner J. , Unger F. 
Published: June 2020

Ultrasonic Aspirators (USA) or Ultrasonic Dissectors, respectively, are used in a variety of surgical disciplines . The functional principle is to fragment and aspi rate tissue components using a piezoelectric PZT (lead, zirconium, tantalum) ceramic electrostrictive transducer tip, which "vibrates" in Tts long axTs at about 23-24 kHz producing longitudinal waves.This study deals with the preparation of the ATI for CABG and compares the effects of conventional preparation of ATI by forceps, scalpel and cautery vs. USA-Application (SONOCORR; SOERING Ltd, Quickborn/FRG), whereby two different energy levels (5%, 25% of the maximal available energy (100W) ) were tested. The aim of the study was to check the effects of the USA on endothelial morphology. Two experiments were done: first, the distal ends of ATIs from four patients immediately prior to CABG were prepared either (two of them) conventionally (cf. above) or by use of USA (one at 5%, one at 25% of max. available energy).
New support film by the plasma polymerization method
Authors: Yamaguchi Masashi , Tanaka Akira , Suzuki Teruo 
Published: June 2020

The plasma polymerization replica method and the apparatus required were deviced by Tanaka. In this technique, ionized gas molecules from a glow discharge are instantly polymerized on the surface of a specimen. The polymerized molecules make a uniform, ultrathin film which precisely copies the three-dimensional ultrastracture of the specimen's surface. In the course of our study, we soon realized that the polymerized film would serve as a good support film. We report here a method of preparation of the new support film by plasma polymerization and its application.Method of preparation: Newly cleaved NaCl or KC1 crystals of 1-2 mm thickness, newly cleaved mica, or coverglass were placed on the cathod plate in a bell jar of the Plasma nano-replica apparatus(PNR-110, Ushio Inc., Tokyo). After evacuation of air, the bell jar was filled with hydrocarbon monomer gas(naphthalene etc.) to about 10 Pa. A high voltage (2-2.5 kV D.C.) was then applied across the electrodes to generate a glow discharge, and a very thin, threedimensionally polymerized film was formed on the surface of substrates.
Electron probe microanalysis of calcium and potassium in cryo-preserved cerebral cortex in situ: A comparison of wet and dry sectioning
Authors: Lightfoot F.G. , Rosenthal R.E. , Fiskum G. 
Published: June 2020

The sub-cellular concentration of calcium and potassium are of particular interest, and have been elusive to investigators not utilizing cryo-ultramicrotomy and cold stage microscopy. By using improved methods in cryo-fixation and tissue drying, one now has an alternate approach to determining ion concentration within the cell. Under certain experimental regimens, metal-mirror fixation is not plausible, and modifications must be adapted.Adult male Beagles, with ischemic brain damage, were used in this study. After the appropriate experimental design of each dog, following anesthesia, a left parietal craniotomy was done and samples of cerebral cortex were removed and processed for biochemical analysis. For analytical electron microscopy (AEM) the dura was removed and the endomarginal gyrus of the parietal lobe was cryofixed using cryo-pliers specifically designed for this procedure. Once the gyrus had been rapidly frozen it was transferred to a cold block for freeze drying under high vacuum (10-5Torr) for 48 hours. After drying, the tissue was fixed for 30 minutes in osmium vapors and infiltrated with Araldite resin and polymerized.
Radial mass analysis of unstained STEM images of molluscan hemocyanins
Authors: Hamilton M.G. , Herskovits T.T. , Wall J.S. 
Published: June 2020

The hemocyanins of molluscs are aggregates of a cylindrical decameric subparticle that assembles into di-, tri-, tetra-, penta-, and larger multi-decameric particles with masses that are multiples of the 4.4 Md decamer. Electron micrographs of these hemocyanins typically show the particles with two profiles: circular representing the cylinder viewed from the end and rectangular representing the side-view of the hollow cylinder.The model proposed by Mellema and Klug from image analysis of a didecameric hemocyanin with the two decamers facing one another with collar (closed) ends outward fits the appearance of side-views of the negatively-stained cylinders. These authors also suggested that there might be caps at the ends. In one of a series of transmission electron microscopic studies of molluscan hemocyanins, Siezen and Van Bruggen supported the Mellema-Klug model, but stated that they had never observed a cap component. With STEM we have tested the end cap hypothesis by direct mass measurements across the end-views of unstained particles.
An integrated screening method for evaluating the pulmonary toxicity of inhaled particulates: Role of microscopic techniques in assessing pulmonary macrophage clearance functions
Authors: Warheit David B. , Achinko Lena , Hartsky Mark A. 
Published: June 2020

There is a great need for the development of a rapid and reliable bioassay to evaluate the pulmonary toxicity of inhaled particles. A number of methods have been proposed, including lung clearance studies, bronchoalveolar lavage analysis, and in vitro cytotoxicity tests. These methods are often limited in scope inasmuch as they measure only one dimension of the pulmonary response to inhaled, instilled or incubated dusts. Accordingly, a comprehensive approach to lung toxicity studies has been developed.To validate the method, rats were exposed for 6 hours or 3 days to various concentrations of either aerosolized alpha quartz silica (Si) or carbonyl iron (CI) particles. Cells and fluids from groups of sham and dust-exposed animals were recovered by bronchoalveolar lavage (BAL). Alkaline phosphatase, LDH and protein values were measured in BAL fluids at several time points postexposure. Cells were counted and evaluated for viability, as well as differential and cytochemical analysis. In addition, pulmonary macrophages (PM) were cultured and studied for morphology, chemotaxis, and phagocytosis by scanning electron microscopy.
SEM of a self-expanding vascular stent in a canine animal model
Authors: Laliberté Verdon S. , Gould K. E. , Termin P. L. , Connolly R. J. , Belkin M. , Breckwoldt W. , Allen M. J. , Hayes L. L. , Callow A. D. 
Published: June 2020

Since the first reported use of a coilspring endovascular prosthesis to improve the outcome of recanalization and ballon angioplasty in peripheral atherosclerotic vessels, a nonthrombogenic, autologous lining has been considered critical for longterm patency. We have investigated the morphology of canine illiac arteries by scanning electron microscopy (SEM) at 3, 7, 21 and 28 days after placement of self-expanding stainless steel stents (Wallstent®).Purpose-bred mongrel dogs, purchased from USDA licensed vendors were housed in the AAALAC approved animal facility at New England Medical Center. Stents (n=18) were placed by percutaneous cannulation into the illiac arteries under fluoroscopic control. The area of each artery containing the stent was excised and perfusion fixed with 3% 0.1M cacodylate buffered glutaraldehyde. Samples were rinsed in 0.1M cacodylate buffer and post-fixed for 60 min. in 0.1M cacodylate buffered 1% Osmium tetroxide. Samples were then rinsed in double-distilled water and fixed for 60 min. in 1% aqueous tannic acid. Samples were dehydrated through graded alcohols, placed in hexamethyldisilazane (HMDS) for 10 minutes and subsequently air dried overnight. Dried specimens were mounted on aluminum stubs with silver paste and examined at 10-20 kv on an Amray 1000B scanning electron microscope.
Preservation and microscopy of bacterial biofilms causing corrosion on steel surfaces
Authors: Aldrich H.C. , McDowell L. , Choate A. , Bono P. 
Published: June 2020

Steel methane transmission lines are sometimes attacked by anaerobic bacteria which cause deep corrosion pits and represent a serious economic problem. We have developed a model laboratory system to follow the process of bacterial attachment and colonization of test specimens of pipeline steel. Test coupons of X and B grades of steel approximately 5mm x 10mm x 20mm were exposed to combinations of the bacteria Enterobacter aerogenes, Desulfovibrio desulfuricans, and Clostridium acetobutylicum in an aerobic culture for periods ranging from 2 hr. to 30 days.A method is described for preserving bacterial films on the surface of the coupons, so that both the bacteria and the corrosion may be studied with light microscopy and scanning and transmission electron microscopy (LM, SEM, TEM). Upon removal from the culture vessel, coupons are fixed in 2.5% glutaraldehyde, dehydrated in ethanols, and embedded in Spurr’s embedding resin, all within a glove bag purged with nitrogen to prevent oxidation of the steel. After polymerization, the plastic is separated by chilling to -80 C. and dropping into hot tap water. The plastic surface next to the metal carries the bacterial film intact, along with impressions of corrosion sites.
Cytochemical study in ischemia injury of renal proximal tubule epithelia cells and the preventive effect of inosine and Ligusticum wallichi
Authors: Zun Zheng , Jihong Sha 
Published: June 2020

The cytochemical methods were used to evaluate the changes of intracellular calcium,plasma meabrane permeability and their relationship during ischemia and reflow in rat renal proximal tubule epithelia cells. In addition, the preventive effects of inosine and Ligusticum Wallichi(LW) on ischemia and reperfusion cell injury were studied and compared.Renal ischemia was induced by 60 min. of renal pedicle clamping in male Spraque-Dawley rats (180-230g) and 2,4, and24 hr. reflow after 60 min. ischemia. The inosine and LW were administed 10 min. and 30 min. respectively before ischemia (Inosine 100mg/100g IV : LW 0.2ml/100g IM;LW is a traditional Chinese medicine, supplied by Chang Hai hospital, Shanghai, China). Calcium was selectively localized with oxalate-pyroantimonate precipitation procedures and analysed by EDX. Lanthanum (La) was employed as a tracer of plasma membrane permeability.Result, Ultrastructural morphology, By the end of 60 min. ischemia, there were no significant differences of cell damage between drug treated groups and untreated groups (Fig.1) but the cell damage would be more and more serious following extension of reflow time.
Ultrastructural localization of immunoglobulin M in activated B-cells in patients with epidemic hemorrhagic fever using gold probe
Authors: Chen Dehui , Guo Ning , Zhang Heqiu , Wang Guohua , Sun Yingxun , Bai Yan 
Published: June 2020

Our previous electron microscopic studies have shown for the first time that the atypical lymphoid cells in the peripheral blood of patients with epidemic hemorrhagic fever (EHF) during the acute febrile phase of the illness appeared to be activated B-cells and plasma cells of various developmental stages. But it is still not a tall clear whether those initially activated cells were actually B-cells. In this study, an indirect immuno gold staining method for the detection of B-cell surface antigen was used.Peripheral blood mononuclear cells were isolated from anticoagulated venous blood of 3 EHF patients (2—7 days of illness) and control donors by centrifugation at 2,500 Xg for 15 min. The cells obtained from the buffy coat were washed in TBS and resuspended in TBS-5% BSA. Unfixed cells were in cubated insuspension in step I with monoclonal antibody (MS27) directed against human IgM (μ Chain) for 30 min at 37±C. After washing in TBS—BSA, the gold probe was applied to the cells in step II incubation with GAM-G15 reagent for 45 min at RT. The gold probe was prepared in our laboratory according to Slot and Geuze. After the third washing in TBS-BSA, the cell pellets were fixed, dehydrated and embedded in Epon 812.
Immunolocalization of Type VI collagen in calfskin: A correlative study
Authors: Kronick Paul L. , Maleeff Beverly E. , Dahms Matthew P. 
Published: June 2020

Although fine details of the fundamental structural units of calfskin, the types I and III collagen fibrils, have been well elucidated, little is known about the location of “minority” collagens in the skin before and after processing to leather. Using microscopic methods, we investigated one such collagen, type VI, in calfskin to determine its presence and distribution.Four different methods of identification of type VI collagen were employed: immunofluorescence and peroxidase-antiperoxidase (PAP) staining using light microscopy, and transmission (TEM) and scanning (SEM) immunoelectron microscopy. In all cases, the primary antibody used was a polyclonal anti-type VI collagen raised in rabbitss. For light microscopy, 12 μm frozen sections were prepared. Fluorescent labeling was carried out with anti-rabbit IgG conjugated to r-phycoerythrin as secondary antibody. PAP staining was performed with the modified techniques of Amenta et al. Tissue for SEM was prepared by a modified protocol of Keene et al. without prefixation: the secondary antibody used was goat anti-rabbit IgG conjugated to 30 nm colloidal gold; it was then fixed in glutaraldehyde-paraformaldehyde and OsO4, dehydrated, critical point dried and sputter coated with gold.
Immunocytochemical mapping of the biosynthetic pathways of two major proteins expressed in terminally differentiated epidermal keratinocytes
Authors: Bisher M.E. , Roop D.R. , Steinert P.M. , Steven A.C. 
Published: June 2020

In the process of constant renewal of mammalian epidermis, cells divide and move upwards from the basal layer as they embark on a program of terminal differentiation. Cells located higher in the epidermis - the spinous, granular, and cornified layers - represent progressively later steps in this pathway. Commitment to terminal differentiation is accompanied by major changes in gene expression and morphology.Inter alia, large-scale synthesis of several proteins is initiated: these include filaggrin, a protein thought to be responsible for aggregating keratin filaments, and a recently discovered protein called loricrin, which has been implicated as a likely major component of the covalently cross-linked protein lining of the cornified cell envelope. We have used immunoelectron microscopy to investigate the biosynthetic pathways of filaggrin and loricrin.Immediately after sacrifice, skin samples were taken from newborn mice, and diced into blocks of 1-2 mm3. They were then either (i) fixed with 1% glutaraldehyde, 0.2% picric acid, in PBS and embedded in LR White, or (ii) fixed with 1% glutaraldehyde in PBS, and embedded in Lowicryl K4M. Essentially the same conclusions were reached from the experiments with both resins.